losartan-potassium and Carcinoma--Renal-Cell

One of the molecules regulated by the transcription factor, hypoxia inducible factor (HIF), is the hypoxia-responsive hematopoietic factor, erythropoietin (EPO). This may have relevance to the development of renal cell carcinoma (RCC), where mutations of the von Hippel-Lindau (VHL) gene are major risk factors for the development of familial and sporadic RCC. VHL mutations up-regulate and stabilize HIF, which in turn activates many downstream molecules, including EPO, that are known to promote angiogenesis, drug resistance, proliferation and progression of solid tumours. HIFs typically respond to hypoxic cellular environment. While the hypoxic microenvironment plays a critical role in the development and progression of tumours in general, it is of special significance in the case of RCC because of the link between VHL, HIF and EPO. EPO and its receptor, EPOR, are expressed in many cancers, including RCC. This limits the use of recombinant human EPO (rhEPO) to treat anaemia in cancer patients, because the rhEPO may be stimulatory to the cancer. EPO may also stimulate epithelial-mesenchymal transition (EMT) in RCC, and pathological EMT has a key role in cancer progression. In this mini review, we summarize the current knowledge of the role of EPO in RCC. The available data, either for or against the use of EPO in RCC patients, are equivocal and insufficient to draw a definitive conclusion.

Topics: Anemia; Animals; Carcinoma, Renal Cell; Erythropoietin; Hematinics; Humans; Hypoxia-Inducible Factor 1; Kidney Neoplasms; Receptors, Erythropoietin; Recombinant Proteins; Risk Factors; Signal Transduction; Tumor Microenvironment; Von Hippel-Lindau Tumor Suppressor Protein

The patient was a 46-year-old man with gross hematuria and left lumbar pain. Computed tomography revealed a large left renal tumor with hemorrhage. Serological examination revealed polycythemia and a high erythropoietin level. After left radical nephrectomy, polycythemia and serologically high level of erythropoietin disappeared. Histopathological findings showed renal cell carcinoma composed of spindle cells. Based on the above mentioned clinical course, we diagnosed this case as erythropoietin-producing renal cell carcinoma with polycythemia. It is suggested that erythropoietin not only causes polycythemia but also stimulates proliferation of the tumor, because the majority of erythropoietin-producing renal cell carcinomas have been reported to be highly advanced. This patient had no evidence of disease one year after the operation.

Topics: Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Neoplasms; Male; Middle Aged; Nephrectomy; Polycythemia

The use of erythropoietin (EPO) for the treatment of anemia associated with urological malignancies is not well defined. The rate of anemia is dependent on the type of cancer and on the different types of treatment. Only with a substantial risk for blood transfusion is substitution treatment by EPO justified. Additionally, the long-term risks of blood transfusions have to be balanced against the costs of EPO treatment.. Different experts have reviewed the literature on anemia and EPO regarding the four main tumor entities.. In prostate cancer, EPO treatment may be justified before radical prostatectomy and in patients with advanced, hormone-refractory disease. In bladder cancer, significant treatment-related anemia mainly occurs in patients who have to undergo radical cystectomy and in patients who will be treated with polychemotherapy for metastatic disease. Patients with renal cell carcinoma rarely suffer from anemia and thus are usually not candidates for EPO treatment. Testis cancer patients only have a substantial risk for blood transfusions if they belong to the intermediate or poor prognosis group according to IGCCCG or if they need salvage chemotherapy or salvage surgery. However, in testis cancer patients EPO treatment should generally be preferred to blood transfusions since cure rates are excellent and thus the potential risks of transfusion-related infections are significant.

Topics: Anemia; Carcinoma, Renal Cell; Erythropoietin; Germinoma; Humans; Male; Testicular Neoplasms; Urologic Neoplasms

Patients with renal cell carcinoma (RCC) rarely develop erythrocytosis. Mechanism of this phenomenon has been disclosed by a series of recent studies using tissue culture technology that erythropoietin (Ep) elaborated by tumor cells. In this review article, the results of in vivo and in vitro studies using the established cell line KU-2 a nude mouse transplantable strain originated from a patient, T.N., with an Ep producing RCC showing erythrocytosis are introduced and discussed on the mechanism of Ep production by the cultured cells. The cultured RCC cells are eligible to produce Ep at stage of over confluence forming the "domes" which are conceivable as the differentiation of the RCC cells in vitro.

Topics: Animals; Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Neoplasms; Mice; Neoplasm Transplantation; Polycythemia; Tumor Cells, Cultured

It is noted that while a wide variety of syndromes have been associated with hypernephroma in the clinical literature, there is clear understanding of the pathophysiology of these effects only in the cases of the endocrine disorders where direct tumor production of hormone can be demonstrated in vitro. Furthermore, this knowledge has done little to alter the care of patients with the disease, except for indications that indomethacin might be of benefit in some patients with hypercalcemia and that one might consider the use of converting enzyme inhibitors in patients with hypernephroma and hypertension. The overall approach to the disease is still surgical. Resection of the tumor also removes the paraneoplastic syndrome. Persistence or recurrence of a syndrome suggests the continued presence of the neoplasm, with the considerations for prognosis which that fact entails. To that degree, at least, these conditions are useful as tumor markers, but such use is limited because they are inconsistent. Further studies of pathophysiology of paraneoplastic syndromes will lead to better understanding of processes of cell differentiation and regulation, and possibly better ways to manage the patients in which they occur.

Topics: Carcinoma, Renal Cell; Chorionic Gonadotropin; Erythropoietin; Glucagon-Like Peptides; Humans; Hypercalcemia; Kidney Neoplasms; Paraneoplastic Endocrine Syndromes; Paraneoplastic Syndromes; Parathyroid Hormone; Prolactin; Prostaglandins; Renin

Hypoxia-inducible factor-2α (HIF-2α) is a transcription factor that frequently accumulates in clear cell renal cell carcinoma (ccRCC), resulting in constitutive activation of genes involved in carcinogenesis. Belzutifan (MK-6482, previously known as PT2977) is a potent, selective small molecule inhibitor of HIF-2α. Maximum tolerated dose, safety, pharmacokinetics, pharmacodynamics and anti-tumor activity of belzutifan were evaluated in this first-in-human phase 1 study (NCT02974738). Patients had advanced solid tumors (dose-escalation cohort) or previously treated advanced ccRCC (dose-expansion cohort). Belzutifan was administered orally using a 3 + 3 dose-escalation design, followed by expansion at the recommended phase 2 dose (RP2D) in patients with ccRCC. In the dose-escalation cohort (n = 43), no dose-limiting toxicities occurred at doses up to 160 mg once daily, and the maximum tolerated dose was not reached; the RP2D was 120 mg once daily. Plasma erythropoietin reductions were observed at all doses; erythropoietin concentrations correlated with plasma concentrations of belzutifan. In patients with ccRCC who received 120 mg once daily (n = 55), the confirmed objective response rate was 25% (all partial responses), and the median progression-free survival was 14.5 months. The most common grade ≥3 adverse events were anemia (27%) and hypoxia (16%). Belzutifan was well tolerated and demonstrated preliminary anti-tumor activity in heavily pre-treated patients, suggesting that HIF-2α inhibition might offer an effective treatment for ccRCC.

Topics: Adult; Aged; Aged, 80 and over; Antineoplastic Agents; Basic Helix-Loop-Helix Transcription Factors; Biomarkers, Tumor; Carcinoma, Renal Cell; Dose-Response Relationship, Drug; Erythropoietin; Female; Humans; Kidney Neoplasms; Male; Middle Aged; Progression-Free Survival; Von Hippel-Lindau Tumor Suppressor Protein

The combination of interferon-alpha2a (IFN-alpha2a) and interleukin-2 (IL-2) induces objective responses in patients with metastatic renal cell carcinoma (MRCC). Anaemia is associated with poor cancer control and reduced quality of life. The aim of the study was to investigate response rate and quality of life in patients with MRCC receiving the combination of Erythropoetin, IFN-alpha2a and IL-2.. Patients with MRCC received epoetin beta (150 IU/kg and haemoglobin <130 g/l or 75 IU/kg and haemoglobin >or=130 g/l) three times weekly, from 14 days before and continuing throughout immunotherapy. In weeks 3-6 the patients received IFN-alpha2a 6 x 10(6) IU/m2 and IL-2 4.5 x 10(6) IU/m2 three times weekly on days 1, 3 and 5. The treatment was repeated two times and in the case of success a third cycle was added. The quality of life was assessed with the FACT questionnaire for fatigue, before, during and after therapy.. A total of 21 patients were treated, 19 of whom could be evaluated concerning response, toxicity and quality of life. We observed 1 complete remission, 2 partial remissions, 5 cases of stable disease and 11 with progressive disease. The overall response rate was 16%. Toxicity was mild to moderate; there were no WHO grade III or IV toxicity. The quality of life increased in ten patients, nine of whom exhibited an increase in their haemoglobin during therapy. Five of the nine patients with decreased quality of life also experienced a decrease in their haemoglobin. The correlation of increased haemoglobin and quality of life was significant (p<0.05).. The combination of IFN-alpha2a, IL-2 and epoetin beta resulted in objective remissions with mild to moderate toxicity. The quality of life correlates significantly with increasing haemoglobin.

Topics: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Bone Neoplasms; Carcinoma, Renal Cell; Chemotherapy, Adjuvant; Combined Modality Therapy; Dose-Response Relationship, Drug; Drug Administration Schedule; Erythropoietin; Female; Follow-Up Studies; Hemoglobinometry; Humans; Injections, Subcutaneous; Interferon alpha-2; Interferon-alpha; Interleukin-2; Kidney Neoplasms; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Nephrectomy; Quality of Life; Recombinant Proteins

Study of the antitumour effects of erythropoietin on metastatic renal cell carcinoma.. After giving their informed consent, 20 patients with histologically proven metastatic renal cell carcinoma received subcutaneous recombinant erythropoietin three times a day at a dose of 150 IU/kg when haemoglobin was less than or equal to 12 g/dL or 75 IU/kg when haemoglobin was higher than 12 g/dL. Treatment was continued for a minimum of 8 weeks before reassessment and was continued thereafter, except in the case of progression or excessive toxicity. A staging assessment was performed every 8 weeks and the response was assessed on the basis of WHO criteria. A clinical and laboratory assessment was performed every two months to evaluate toxicity, graded according to the WHO scale. All but one of the patients had received immunotherapy or chemotherapy prior to inclusion in the study.. One complete response (> 12 months), one partial response (8 months), two minor responses, 10 cases of stabilisation and 6 cases of progression were observed. 15 patients received treatment at full doses. In 5 patients, the duration of treatment was reduced before the 8 weeks initially defined because of tumour progression in one patient and because of haemoglobin persistently greater than 15 g/dL in 4 other patients. Adverse effects consisted of 1 case of moderate headache, 2 cases of transient bone pain, and 1 case of transient hypertension.. Erythropoietin exerts a moderate antitumour effect which needs to be confirmed by a phase II trial of first-line treatment in selected patients.

Topics: Aged; Antineoplastic Agents, Hormonal; Bone and Bones; Carcinoma, Renal Cell; Disease Progression; Erythropoietin; Female; Headache; Hemoglobins; Humans; Hypertension; Injections, Subcutaneous; Kidney Neoplasms; Male; Middle Aged; Neoplasm Staging; Pain; Prospective Studies; Recombinant Proteins; Remission Induction; World Health Organization

Topics: Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Diseases, Cystic; Kidney Neoplasms; Polycythemia

Topics: Aged; Carcinoma, Renal Cell; Diagnosis, Differential; Dizziness; Erythropoietin; Fatigue; Humans; Kidney Neoplasms; Lung Neoplasms; Male; Nephrectomy; Polycythemia; Tomography, X-Ray Computed; Treatment Outcome

In this study, we report here a rare case of polycythemia and cRCC in the same patient, which may be helpful in understanding clinical features and molecular mechanisms underlying VHL-mutation-associated cRCC and polycythemia induced by germline mutation of HIF2A. Firstly, we identified a rare but well studied germline mutation resulting in polycythemia in HIF2A (c.1609G>A, p.Gly537Arg) in the blood of the patient and his daughter. Meanwhile, we identified an inactivating VHL mutation (c.391A>T, p.N131Y), as well as TP53 mutation(c.977A>T, p.E326V) and mTOR mutation(c.7498A>T, p.I2500F) in renal cancer tissue. Moreover, protein levels of VHL, HIF1A, HIF2A, EPO, and VEGF estimated by immunohistochemical staining substantiated hyperactivation of the oxygen-sensing pathway. In addition, we identified 158 somatic SNP/indel mutations, including 90 missense/nonsense/splice/stop-loss mutations by whole-exome sequencing (WES) of the tumor specimen and matched normal DNA.

Topics: Basic Helix-Loop-Helix Transcription Factors; Carcinoma, Renal Cell; DNA Mutational Analysis; Erythropoietin; Germ-Line Mutation; Humans; Male; Middle Aged; Mutation; Polycythemia; Polymorphism, Single Nucleotide; TOR Serine-Threonine Kinases; Tumor Suppressor Protein p53; Von Hippel-Lindau Tumor Suppressor Protein

The aim of the study was to investigate the expression of erythropoietin and neuroendocrine markers in clear cell renal cell carcinoma (CCRCC). We retrospectively reviewed the medical records and re-evaluated histopathological specimens of 33 patients with CCRCC and compared with those of 11 cases of non-CCRCC. All patients were treated with a partial or radical nephrectomy at St. Olavs Hospital, Trondheim University Hospital, between 2010 and 2016. Thirty-three patients who were diagnosed with CCRCC had a total of 35 tumours, where 34 of the tumours were CCRCC and one was papillary adenoma. Thirty-three (97%) of 34 CCRCCs were positive for erythropoietin, and the same 33 (97%) tumours demonstrated strong expression for neuron-specific enolase (NSE). Two (6%) of 34 CCRCCs had a positive reaction for synaptophysin, and three (9%) of 34 were positive for CD56. Erythropoietin and NSE were negative in non-CCRCCs, and chromogranin A was negative in all tumours. The above findings suggest that there is a strong association between CCRCC and the expression of erythropoietin and NSE.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Renal Cell; CD56 Antigen; Erythropoietin; Female; Humans; Immunohistochemistry; In Situ Hybridization; Kidney Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Retrospective Studies; Synaptophysin

Erythropoietin (EPO) is a frequently prescribed anti-anemic drug for patients with advanced renal carcinoma. However, recent evidence from clinical studies suggested that EPO accelerated tumor progression and jeopardized the 5-year survival. Herein, we show, starting from the in silico microarray bioinformatics analysis, that activation of Erythropoietin signaling pathway enhanced renal clear carcinoma (RCC) progression. EPO accelerated the proliferative and migratory ability in 786-O and Caki-2 cells. Moreover, comparative proteomics expression profiling suggested that exogenous EPO stimulated RCC progression via up-regulation of KIAA0101 expression. Loss of KIAA0101 impeded the undesirable propensity of EPO in RCC. Finally, low expression of KIAA0101 was associated with the excellent prognosis and prognosticated a higher 5-year survival in human patients with renal carcinoma. Overall, KIAA0101 appears to be a key promoter of RCC malignancy induced by EPO, which provide mechanistic insights into KIAA0101 functions, and pave the road to develop new therapeutics for treatment of cancer-related and chemotherapy-induced anemia in patients with RCC.

Topics: Apoptosis; Biomarkers, Tumor; Carcinoma, Renal Cell; Carrier Proteins; Cell Movement; Cell Proliferation; DNA-Binding Proteins; Erythropoietin; Humans; Kidney Neoplasms; Neoplasm Staging; Prognosis; Proteomics; Survival Rate; Tumor Cells, Cultured

This article seeks to clarify if gender-based differences occur in the pharmacokinetics of metoprolol in the elderly patients. There are a series of physiologic changes that occur in the elderly ranging from decreased hepatic blood flow to increased adiposity causing higher plasma concentrations at therapeutic doses as compared to the healthy young population.. Population pharmacokinetic modeling were performed using MONOLIX and Monte-Carlo simulations were conducted using MATLAB. The data was based from a previously published dataset where elderly patients, having multiple comorbidities, were administered a 50mg dose of metoprolol.. Gender stratified doses resulting in an equivalent systemic metoprolol exposure in geriatric patients have been identified. Metoprolol doses resulting a similar AUC in a healthy young male administered 50mg tablet were 15mg for geriatric women and 25mg for geriatric men. Further, Metoprolol doses of 25mg for geriatric women and 50mg for geriatric men resulted in an equivalent AUC to a healthy young males dosed with a 100mg tablet. A 15mg Metoprolol tablet may need to be compounded to account for the gender differences in Metoprolol pharmacokinetics.

Topics: Adult; Aged; Aged, 80 and over; Animals; Antigens, Ly; Apoptosis; Astrocytes; Biomarkers; Biomarkers, Tumor; Blotting, Western; Bone Marrow Cells; Brain Ischemia; Bromodeoxyuridine; Carcinoma, Renal Cell; Case-Control Studies; Cell Hypoxia; Cell Movement; Cell Proliferation; Cells, Cultured; Chemokines; Diabetes, Gestational; Down-Regulation; Erythropoietin; Female; Gestational Age; Glucose; Heme Oxygenase-1; Heterozygote; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunoenzyme Techniques; Insulin-Like Growth Factor Binding Protein 1; Insulin-Like Growth Factor Binding Protein 2; Insulin-Like Growth Factor Binding Protein 3; Insulin-Like Growth Factor I; Kidney Neoplasms; Longitudinal Studies; Male; Mice, Inbred C57BL; Mice, Knockout; Middle Aged; Mitogen-Activated Protein Kinases; Myeloid Cells; Neoplasm Grading; Neoplasm Staging; Neuroprotection; Odds Ratio; Oxidation-Reduction; Oxygen; Pregnancy; Prognosis; Rats; Real-Time Polymerase Chain Reaction; Receptors, CCR2; Receptors, Erythropoietin; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; RNA, Messenger; Signal Transduction; Stearoyl-CoA Desaturase; Survival Rate; TRPV Cation Channels; Tumor Cells, Cultured; Uterus; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-2; Young Adult

Clear cell renal cell carcinoma (ccRCC) is characterized by inactivation of the von Hippel-Lindau tumour suppressor gene (VHL). Because no other gene is mutated as frequently in ccRCC and VHL mutations are truncal, VHL inactivation is regarded as the governing event. VHL loss activates the HIF-2 transcription factor, and constitutive HIF-2 activity restores tumorigenesis in VHL-reconstituted ccRCC cells. HIF-2 has been implicated in angiogenesis and multiple other processes, but angiogenesis is the main target of drugs such as the tyrosine kinase inhibitor sunitinib. HIF-2 has been regarded as undruggable. Here we use a tumourgraft/patient-derived xenograft platform to evaluate PT2399, a selective HIF-2 antagonist that was identified using a structure-based design approach. PT2399 dissociated HIF-2 (an obligatory heterodimer of HIF-2α-HIF-1β) in human ccRCC cells and suppressed tumorigenesis in 56% (10 out of 18) of such lines. PT2399 had greater activity than sunitinib, was active in sunitinib-progressing tumours, and was better tolerated. Unexpectedly, some VHL-mutant ccRCCs were resistant to PT2399. Resistance occurred despite HIF-2 dissociation in tumours and evidence of Hif-2 inhibition in the mouse, as determined by suppression of circulating erythropoietin, a HIF-2 target and possible pharmacodynamic marker. We identified a HIF-2-dependent gene signature in sensitive tumours. Gene expression was largely unaffected by PT2399 in resistant tumours, illustrating the specificity of the drug. Sensitive tumours exhibited a distinguishing gene expression signature and generally higher levels of HIF-2α. Prolonged PT2399 treatment led to resistance. We identified binding site and second site suppressor mutations in HIF-2α and HIF-1β, respectively. Both mutations preserved HIF-2 dimers despite treatment with PT2399. Finally, an extensively pretreated patient whose tumour had given rise to a sensitive tumourgraft showed disease control for more than 11 months when treated with a close analogue of PT2399, PT2385. We validate HIF-2 as a target in ccRCC, show that some ccRCCs are HIF-2 independent, and set the stage for biomarker-driven clinical trials.

Topics: Animals; Aryl Hydrocarbon Receptor Nuclear Translocator; Basic Helix-Loop-Helix Transcription Factors; Binding Sites; Carcinoma, Renal Cell; Cell Line, Tumor; Cell Transformation, Neoplastic; Drug Resistance, Neoplasm; Erythropoietin; Female; Gene Expression Regulation, Neoplastic; Humans; Indans; Indoles; Kidney Neoplasms; Male; Mice; Mice, Inbred NOD; Mice, SCID; Molecular Targeted Therapy; Mutation; Pyrroles; Reproducibility of Results; Sulfones; Sunitinib; Xenograft Model Antitumor Assays

To investigate the correlation between parathyroid hormone-related protein (PTHrP), erythropoietin (EPO), and vascular endothelial growth factor (VEGF) expression in clear cell renal cell carcinoma (ccRCC). Immunohistochemical studies on PTHrP, EPO and VEGF were performed in 249 patients with ccRCC. Serum calcium level and haematocrit were analyzed. The expression of the factors and clinicopathological parameters were studied statistically for possible correlations. The incidence for hypercalcaemia and polycythaemia were 15.3% and 2.0% respectively. Expression of PTHrP, EPO, and VEGF were respectively related to advanced stage (P < 0.0001 respectively). PTHrP was not related to tumour grade. Expressions of EPO and VEGF were correlated to tumour grade significantly. All factors were expressed higher in hypercalcaemic patients. PTHrP, EPO, and VEGF were positively correlated with each other in non-hypercalcaemic patients yet not in hypercalcaemic ones. PTHrP and EPO are related to VEGF expression and to the progression of ccRCC. This finding offers us new insight on the behaviour of ccRCC and offers possible targets in RCC treatment.

Topics: Calcium; Carcinoma, Renal Cell; Cohort Studies; Disease Progression; Erythropoietin; Female; Humans; Hypercalcemia; Immunohistochemistry; Kidney Neoplasms; Male; Neoplasm Grading; Neoplasm Staging; Parathyroid Hormone-Related Protein; Vascular Endothelial Growth Factor A

Erythropoietin (EPO) provides an alternative to transfusion for increasing red blood cell mass and treating anemia in cancer patients. However, recent studies have reported increased adverse events and/or reduced survival in patients receiving both EPO and chemotherapy, potentially related to EPO-induced cancer progression. Additional preclinical studies that elucidate the possible mechanism underlying EPO cellular growth stimulation are needed.. Using commercial tissue microarray (TMA) of a variety of cancers and benign tissues, EPO and EPO receptor immunohistochemical staining was performed. Furthermore using a panel of human renal cells (Caki-1, 786-O, 769-P, RPTEC), in vitro and in vivo experiments were performed with the addition of EPO in normoxic and hypoxic states to note phenotypic and genotypic changes.. EPO expression score was significantly elevated in lung cancer and lymphoma (compared to benign tissues), while EPOR expression score was significantly elevated in lymphoma, thyroid, uterine, lung and prostate cancers (compared to benign tissues). EPO and EPOR expression scores in RCC and benign renal tissue were not significantly different. Experimentally, we show that exposure of human renal cells to recombinant EPO (rhEPO) induces cellular proliferation, which we report for the first time, is further enhanced in a hypoxic state. Mechanistic investigations revealed that EPO stimulates the expression of cyclin D1 while inhibiting the expression of p21cip1 and p27kip1 through the phosphorylation of JAK2 and ERK1/2, leading to a more rapid progression through the cell cycle. We also demonstrate an increase in the growth of renal cell carcinoma xenograft tumors when systemic rhEPO is administered.. In summary, we elucidated a previously unidentified mechanism by which EPO administration regulates progression through the cell cycle, and show that EPO effects are significantly enhanced under hypoxic conditions.

Topics: Animals; Carcinoma, Renal Cell; Cell Cycle; Cell Growth Processes; Cell Hypoxia; Cell Line, Tumor; Erythropoietin; Heterografts; Humans; Immunohistochemistry; Janus Kinase 2; Kidney Neoplasms; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Mice, Nude; Random Allocation; Receptors, Erythropoietin; Recombinant Proteins; Signal Transduction

Co-expression of erythropoietin (Epo) and erythropoietin receptor (EpoR) has been found in various non-hematopoietic cancers including hereditary and sporadic renal cell carcinomas (RCC), but the Epo/EpoR autocrine and paracrine mechanisms in tumor progression have not yet been identified. In this study, we used RNA interference method to down-regulate EpoR to investigate the function of Epo/EpoR pathway in human RCC cells. Epo and EpoR co-expressed in primary renal cancer cells and 6 human RCC cell lines. EpoR signaling was constitutionally phosphorylated in primary renal cancer cells, 786-0 and Caki-1 cells, and recombinant human Epo (rhEpo) stimulation had no significant effects on further phosphorylation of EpoR pathway, proliferation, and invasiveness of the cells. Down-regulation of EpoR expression in 786-0 cells by lentivirus-introduced siRNA resulted in inhibition of growth and invasiveness in vitro and in vivo, and promotion of cell apoptosis. In addition, rhEpo stimulation slightly antagonized the anti-tumor effect of Sunitinib on 786-0 cells. Sunitinib could induce more apoptotic cells in 786-0 cells with knockdown EpoR expression. Our results suggested that Epo/EpoR pathway was involved in cell growth, invasion, survival, and sensitivity to the multi-kinases inhibitor Sunitinib in RCC cells.

Topics: Animals; Apoptosis; Carcinoma, Renal Cell; Cell Line, Tumor; Cell Proliferation; Erythropoietin; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Indoles; Kidney Neoplasms; Matrix Metalloproteinase 2; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Protein Kinase Inhibitors; Pyrroles; Receptors, Erythropoietin; Recombinant Proteins; Signal Transduction; Sunitinib

• To elucidate the association of progression of advanced renal cell carcinoma with anaemia and investigate factors influencing tumor-associated anaemia.. • We analyzed different clinical variables to study associations with anaemia in 86 metastatic renal cell carcinoma patients. • 45 (52%) of patients had already developed anaemia prior to therapy.. • Anaemic patients had an increase in the serum markers C-reactive protein (CRP), IL-6 and erythropoietin (EPO). In addition we observed substantial correlation between IL-6 and CRP serum levels (R = 0.639, P < 0.0001). • Univariate logistic regression analysis revealed that patients with IL-6 >10 pg/mL had a considerable increase in risk for anaemia (odds ratio 3.86, P= 0.003). • In addition, patients with CRP >0.7 mg/dL had a very strong increase in risk for anaemia (OR = 14.08, P < 0.0001). • Stepwise multivariate logistic regression analysis confirmed CRP >0.7 mg/mL as the only independent predictor for anaemia. Cox-regression modeling selected serum IL-6 as the strongest independent prognostic indicator (hazard ratio 3.58, P < 0.0001).. • Anaemia depends on serum IL-6, which is a strong inductor of CRP and regulator of the iron-transport. Serum IL-6 may be considered as a target to treat cancer-related anaemia.

Topics: Aged; Anemia; Biomarkers; C-Reactive Protein; Carcinoma, Renal Cell; Epidemiologic Methods; Erythropoietin; Female; Humans; Interleukin-6; Kidney Neoplasms; Male; Middle Aged; Prognosis

We investigated the safety and feasibility of sorafenib in patients with end-stage renal disease undergoing hemodialysis by examining the influence of pharmacokinetic parameters to their benefit and also the occurrence of drug-related adverse events of sorafenib.. Ten patients with metastatic renal cell carcinoma undergoing hemodialysis received sorafenib. Initial dose was 200 mg once daily, and the dose was increased up to the maintenance dose of 200 mg twice daily. The pharmacokinetic study was performed after a steady state was reached with 200 mg twice daily in six patients.. Complete response occurred in one patient, partial response in three, stable disease in four and progressive disease in two. Median progression-free survival was 6.3 months. Serious adverse events were found in nine patients, including a Grade 5 subarachnoid hemorrhage and a Grade 4 cerebellar hemorrhage. In the pharmacokinetic study, the geometric mean of maximum concentration and area under the curve from 0 to 10 h of plasma concentration were similar on the day of hemodialysis and the day off hemodialysis. These data were lower than those from Japanese people with healthy kidneys and normal kidney function. There was no association between objective response or the occurrence of serious adverse events and pharmacokinetic parameters.. Treatment with sorafenib of patients with metastatic renal cell carcinoma undergoing hemodialysis appears to be feasible, but we express some concern about the higher incidence of serious adverse events even with the reduced dose. However, clinical efficacy was not compromised.

Topics: Aged; Antineoplastic Agents; Benzenesulfonates; Carcinoma, Renal Cell; Disease-Free Survival; Drug Administration Schedule; Erythropoietin; Feasibility Studies; Hematinics; Hemoglobins; Humans; Kaplan-Meier Estimate; Kidney Failure, Chronic; Kidney Neoplasms; Male; Middle Aged; Niacinamide; Phenylurea Compounds; Protein Kinase Inhibitors; Pyridines; Renal Dialysis; Sample Size; Sorafenib; Treatment Outcome

Decreased levels of von Hippel-Lindau (VHL) tumour suppressor protein are associated with up-regulation of hypoxia-inducible factor (HIF), leading to increased tumour proliferation, angiogenesis and progression. The role of erythropoietin (EPO), a target gene for HIF, remains unknown for sporadic clear cell renal cell carcinoma (sCCRCC). In this study, we determined expression levels of EPO, and its correlation with VHL mutations and HIF-1α and HIF-2α expression in 82 patients identified with sCCRCC following nephrectomy. We identified VHL gene alterations using multiplex polymerase chain reaction, purifying products of polymerase chain reaction, and direct sequencing. Immunohistochemical staining for HIF-1α, HIF-2α and EPO was performed for tumour and corresponding normal tissues. Data were analyzed with respect to clinicopathological factors. EPO was detected in 87.8% of sCCRCC tumours versus 7.3% for normal tissues. EPO expression was related to tumours demonstrating VHL gene abnormalities. Of specimens with VHL alterations 95.6% tested positive for EPO, versus 78.3% when VHL gene expression was normal (P<0.01). EPO was identified in 96.2 and 94.2% of HIF-1α and HIF-2α positive specimens, respectively, compared to 72.4 and 53.8% for HIF-1α and HIF-2α negative groups (p<0.01). Moreover, EPO expression correlated significantly with increasing nuclear grade (p<0.05). HIF-2α was identified in 84.1% of sCCRCC, compared to 64.6% for HIF-1α. Expression of HIF-1α, HIF-2α and EPO is common in sCCRCC. Although both forms of HIF up-regulate expression of EPO, the relationship to HIF-2α appears to be more pronounced. The VHL-HIF-EPO pathway requires further study, as it may represent a potential molecular target for therapy of sCCRCC.

Topics: Basic Helix-Loop-Helix Transcription Factors; Carcinoma, Renal Cell; Chi-Square Distribution; DNA Methylation; Electrophoresis, Agar Gel; Erythropoietin; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunohistochemistry; Kidney Neoplasms; Mutation; Neoplasm Staging; Von Hippel-Lindau Tumor Suppressor Protein

Topics: Antineoplastic Agents; Axitinib; Carcinoma, Renal Cell; Erythropoietin; Humans; Imidazoles; Indazoles; Kidney Neoplasms; Male; Middle Aged; Protein-Tyrosine Kinases

Mature circulating endothelial cell (CEC) as well as endothelial progenitor populations may reflect the activity of anti-angiogenic agents on tumor neovasculature or even constitute a target for anti-angiogenic therapy. We investigated the behavior of CECs in parallel with hematopoietic progenitor cells (HPCs) in the blood of renal cell cancer patients during sunitinib treatment. We analyzed the kinetics of a specific population of small VEGFR2-expressing CECs (CD45(neg)/CD34(bright)), HPCs (CD45(dim)/CD34(bright)), and monocytes in the blood of 24 renal cell cancer (RCC) patients receiving 50 mg/day of the multitargeted VEGF inhibitor sunitinib, on a 4-week-on/2-week-off schedule. Blood was taken before treatment (C1D1), on C1D14, C1D28, and on C2D1 before the start of cycle 2. Also plasma VEGF and erythropoietin (EPO) were determined. Remarkably, while CD34(bright) HPCs and monocytes decreased during treatment, CD34(bright) CECs increased from 69 cells/ml (C1D1) to 180 cells/ml (C1D14; P = 0.001) and remained high on C1D28. All cell populations recovered to near pre-treatment levels on C2D1. Plasma VEGF and EPO levels were increased on C1D14 and partly normalized to pre-treatment levels on C2D1. In conclusion, opposite kinetics of two circulating CD34(bright) cell populations, HPCs and small CECs, were observed in sunitinib-treated RCC patients. The increase in CECs is likely caused by sunitinib targeting of immature tumor vessels.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD34; Antineoplastic Agents; Biomarkers, Tumor; Blood Cell Count; Carcinoma, Renal Cell; Endothelial Cells; Erythropoietin; Female; Flow Cytometry; Hematopoietic Stem Cells; Humans; Indoles; Kidney Neoplasms; Kinetics; Male; Middle Aged; Pyrroles; Sunitinib; Treatment Outcome; Vascular Endothelial Growth Factor A

To investigate the prognostic impact of erythropoietin (EPO) and EPO-receptor (EPO-R) expression in tumour as well as serum EPO in patients with renal cell carcinoma (RCC).. Using immunohistochemistry, EPO and EPO-R were assessed in tissue microarrays from 195 RCCs. RCC type, TNM stage, nuclear grade, survival, EPO and haemoglobin (Hb) levels in blood were registered.. Strong expression of EPO and EPO-R in tumour tissue was found in 83 and 56%, respectively. EPO and EPO-R expression differed between RCC types. Serum EPO and blood Hb did not correlate to the expression of EPO or EPO-R. A positive correlation was found between the expression of EPO and EPO-R (p = 0.028). Survival was not related to tumour EPO, whereas strong EPO-R expression indicated a non-significantly worse prognosis. Serum EPO correlated positively to TNM stage and nuclear grade and negatively to survival. A multivariate analysis showed that TNM stage and nuclear grade were independent prognostic factors. Tumour EPO and EPO-R expression as well as serum EPO added no independent prognostic information.. No correlation between EPO or EPO-R in tumour tissue and serum EPO or blood Hb was found. Neither EPO, EPO-R in tumour tissue nor serum EPO are independent prognostic factors.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Renal Cell; Erythropoietin; Female; Gene Expression; Hemoglobins; Humans; Male; Middle Aged; Neoplasm Staging; Prognosis; Receptors, Erythropoietin; Tissue Array Analysis

Mutation of the VHL tumor suppressor gene is a frequent genetic event in the carcinogenesis of renal-cell carcinoma (RCC). Circulating endothelial progenitor cells (EPCs) have important role in neoangiogenesis, and mobilization of these cells is induced by various growth factors including erythropoietin (EPO). With this regard, we analyzed a patient with EPO-producing clear-cell RCC and polycythemia. DNA extraction and sequencing analysis of the VHL gene were performed from the tumor and the adjacent normal renal tissue. Isolated and cultured circulating EPCs from the blood taken with phlebotomy were characterized by flow cytometry and immunofluorescence analysis. This RCC had two novel somatic mutations of the VHL gene, p.Leu128Pro and p.Asn131Lys. Culture of blood mononuclear cells revealed a strikingly high number of endothelial cell colonies derived from EPCs (nearly 10-fold more than in controls). Elevated number of circulating EPCs seems to be related to high EPO production from RCC with novel double somatic mutation of the VHL gene in this patient.

Topics: Aged; Carcinoma, Renal Cell; DNA, Neoplasm; Endothelium, Vascular; Erythropoietin; Humans; Kidney Neoplasms; Male; Mutation; Nephrectomy; Polycythemia; Reverse Transcriptase Polymerase Chain Reaction; Stem Cells; Von Hippel-Lindau Tumor Suppressor Protein

The treatment of metastatic renal cell carcinoma (RCC) remains a clinical challenge. Factors predicting any benefit of different therapies would therefore be useful. Angiogenesis is important in tumor progression and the development of metastases. The aim of this study in patients with distant metastases at diagnosis was to evaluate possible outcome information obtained with a number of soluble angiogenic variables in serum.. Serum samples were taken at diagnosis from 120 consecutive patients with metastatic RCC who were operated on with radical nephrectomy. Different clinicopathological variables and vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor-1 (VEGFR-1), basic fibroblast growth factor (bFGF) and erythropoietin levels in serum were compared with the clinical course.. The median survival time for all patients was 9 months. Six patients (5%) died during the postoperative period, all of whom had a performance status (PS) of 2 or 3. None of the angiogenic factors (VEGF, VEGFR-1, bFGF, erythropoietin) gave any prognostic information, except that VEGF was associated with survival (p = 0.0234) in patients with a good PS. A number of other variables gave prognostic information in univariate analysis but, after multivariate analysis, only PS (p = 0.002), the number of metastatic sites (p = 0.003) and capsule invasion (p = 0.017) remained as independent predictive factors.. Among predictive factors, only PS, the number of metastatic sites and capsule invasion independently predicted survival in patients with metastatic RCC, while soluble angiogenic factors in serum gave no prognostic information. Nephrectomy in patients with metastatic RCC remains controversial but long-term survival can be achieved in selected patients, especially those with a good PS.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Renal Cell; Disease Progression; Erythropoietin; Female; Fibroblast Growth Factor 2; Humans; Kidney Neoplasms; Male; Middle Aged; Neovascularization, Pathologic; Prognosis; Proportional Hazards Models; Survival Analysis; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1

To investigate, in a retrospective study, the expression of erythropoietin (Epo) in human renal cell carcinoma (RCC) and its correlation with overall survival, as Epo (an haematopoietic cytokine that regulates the production of red blood cells), with its receptor, was recently localized in non-haematopoietic tissues, e.g. liver, uterus, central nervous system, vascular endothelial cells and solid tumours.. We used data from 113 patients who had radical nephrectomy for RCC between 1990 and 2000, taking sections from formalin-fixed and paraffin wax-embedded tissue blocks. The association between Epo staining and the patients' characteristics was assessed by either chi-squared tests (for categorical variables) or two-sample independent t-tests (for continuous variables).. Tissue from 37 patients (33%) was positive for cytoplasmic Epo expression; 76 (67%) samples were negative. Univariate hazard ratio analysis confirmed that those with positive Epo staining were more than twice as likely to die as those with negative staining (hazard ratio 2.34, 95% confidence interval 1.27-4.3).. This study shows that the expression of Epo in RCC is adversely associated with overall survival. This is the first report of such an association, and might be explained by the loss of Von Hippel-Lindau protein function in clear cell RCC. The expression of Epo might have potential use in clinical trials when stratifying high-risk patients for adjuvant therapy after nephrectomy.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Immunohistochemistry; Kidney Neoplasms; Male; Middle Aged; Multivariate Analysis; Nephrectomy; Prognosis; Retrospective Studies; Survival Analysis

Signalling by erythropoietin (EPO) is increasingly recognised as a relevant mechanism in tumour biology, potentially leading to enhanced proliferation, angiogenesis and therapy resistance. Paraneoplastic polycythemia by cancerous overproduction of EPO is a rare event, but most frequently seen in patients with renal cell carcinoma (RCC). The majority of clear cell RCC displays a strong activation of the transcription factor regulating EPO, the Hypoxia-inducible Factor (HIF). Therefore, it is unclear why only a small minority of patients develop polycythemia. We studied 70 RCC for EPO gene and HIFalpha isoform expression. 34% of all RCC showed expression of EPO mRNA in RNase protection assays, which were almost exclusively of the clear cell type. Only 1 patient presented with polycythemia. In situ hybridisation revealed that expression of EPO was in the tumour cells. Expression of EPO mRNA was always associated with activation of HIF, which could involve HIF-1alpha and/or HIF-2alpha. The frequency of EPO gene expression in RCC is therefore much higher than the prevalence of polycythemia. Furthermore, activation of HIF appears necessary for EPO gene expression in RCC, but is clearly not the only determinant. Further to the reported expression of EPO receptors in tumour tissues, the finding of widespread expression of EPO in RCC supports the recent notion of an involvement of this system in paracrine or autocrine effects of tumour cells.

Topics: Adenocarcinoma, Clear Cell; Basic Helix-Loop-Helix Transcription Factors; Carcinoma, Renal Cell; Cell Line, Tumor; Erythropoietin; Gene Expression Regulation, Neoplastic; Germany; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunoblotting; In Situ Hybridization; Kidney Neoplasms; Paraneoplastic Syndromes; Polycythemia; Prevalence; Ribonucleases; RNA, Messenger; Signal Transduction; Tumor Cells, Cultured; Up-Regulation

Anaemia which develops as a consequence of malignancies is often treated using recombinant human erythropoietin (rhEpo). Epo is now known as an anti-apoptotic factor for a wide range of cell types that express Epo receptors (EpoRs) and its co-use with cancer therapies can act detrimentally to diminish therapy-induced apoptosis. This had not been analyzed for renal cell carcinomas (RCCs). We examined the influence of rhEPO on the ability of cisplatin to induce apoptosis in RCCs. Two RCC cell lines (SN12K1 and ACHN) were compared with a non-RCC renal epithelial cell line (HK2). Cells were treated with 50 microM cisplatin with and without 200 IU/mL rhEpo and were compared for apoptosis, mitosis and protein expression of EpoR, nuclear factor-kappaB (NFkappaB), protein kinase C (PKC), Bcl-2, Bax and cyclin-D1. Experiments were repeated with PKC promotion (PMA, 20 nM) or inhibition (H7, 10 microM). rhEpo reduced cisplatin-induced apoptosis in RCCs (p < 0.01), compared with HK-2s. EpoR expression was increased only in SN12K1 with rhEpo, with and without cisplatin. NFkappaB, Bax and Bcl-2 expression was unchanged. PKC protein expression was significantly reduced in cisplatin-treated RCCs with rhEpo, correlating with reduced apoptosis. When the PKC pathway was inhibited in these cells, levels o apoptosis returned to normal for cisplatin treatment, indicating activation of the PKC pathway by rhEpo. PMA promotion increased mitosis only in the RCCs, with and without rhEpo (p < 0.05). In summary, rhEPO reduced cisplatin-induced apoptosis of RCCs and promoted their mitosis via PKC-dependent pathways. This information indicates caution for use of rhEpo in RCC patients for anemias.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Anemia; Apoptosis; Carcinoma, Renal Cell; Cell Line, Tumor; Cisplatin; Enzyme Activation; Epithelial Cells; Erythropoietin; Humans; Kidney Neoplasms; Mitosis; Neoplasm Proteins; Protein Kinase C; Recombinant Proteins; Signal Transduction; Tetradecanoylphorbol Acetate

Clear cell renal cell carcinoma (CCRCC) is the most common renal carcinoma and it is often associated with von Hippel-Lindau disease (VHL) gene mutations. CCRCCs with VHL mutations demonstrate hypoxia-inducible factor (HIF) overexpression as well as increased expression of vascular endothelial growth factor (VEGF). Recently, the erythropoietin (Epo) has been found to be upregulated in renal and other tumors associated with VHL disease. Furthermore, Epo and Epo receptor (EpoR) coexpression has also been reported in these tumors. The results provided strong evidence that an autocrine loop is involved in tumorigenesis in VHL disease. We investigated whether Epo and EpoR coexpression also occurs in sporadic CCRCC. Fifty-four sporadic CCRCCs were analyzed. VHL gene mutations were detected in 30 out of 54 tumors. Coexpression of Epo and EpoR was detected in 50 out of 54 tumors regardless of their VHL mutation status. The results suggest that coexpression of Epo and EpoR plays an important role in tumorigenesis of sporadic CCRCC.

Topics: Adenocarcinoma, Clear Cell; Amino Acid Substitution; Base Sequence; Carcinoma, Renal Cell; Codon; DNA Primers; Erythropoietin; Humans; Kidney Neoplasms; Receptors, Erythropoietin; Reverse Transcriptase Polymerase Chain Reaction

Metastatic renal cell carcinoma (MRCC) involving the thorax can be difficult to distinguish from diffuse malignant mesothelioma (DMM) using traditional morphologic approaches. Standard panels of immunohistochemical markers are of limited benefit.. To investigate several antibodies to renal cell carcinoma-associated proteins for differentiating MRCC from DMM.. One hundred DMMs and 20 MRCCs were evaluated for immunoexpression of erythropoietin. The same cases and an additional 45 DMMs were evaluated for CD10 and renal cell carcinoma marker (RCCMa) immunoreactivity.. Erythropoietin was expressed in 100% of DMMs and MRCCs. Staining for CD10 was observed in 54% of DMMs and 100% of MRCCs. RCCMa stained 26% of DMMs and 55% of MRCCs. Although erythropoietin staining was similarly strong and diffuse in both DMM and MRC, patterns of staining for RCCMa and CD10 differed between MRCC and DMM. Immunoreactivity was strong and diffuse for both RCCMa and CD10 in most MRCCs. Of CD10-positive DMMs, nearly half showed staining in less than 50% of tumor cells and about one fourth of positive cases exhibited only weak to moderately intense staining. Only half of RCCMa-positive DMMs showed staining in more than 49% of tumor cells and staining was only weak to moderately intense in most cases.. Given the overlap in the expression of renal cell carcinoma markers in MRCC and DMM, results with these markers must be interpreted cautiously and should be used in conjunction with mesothelium-associated markers. Differences in expression may potentially help distinguish MRCC from DMM inasmuch as strong and diffuse expression of RCCMa and CD10 supports a diagnosis of MRCC over DMM.

Topics: Biomarkers, Tumor; Carcinoma, Renal Cell; Diagnosis, Differential; Erythropoietin; Humans; Immunoenzyme Techniques; Kidney Neoplasms; Mesothelioma; Neprilysin; Pleural Neoplasms

Two cell lines that exemplify erythropoietin (EPO) receptor-positive tumors, human renal carcinoma cell lines RCC and the myelomonocytic leukemia cell line U937, were investigated for the apoptosis-modulatory potential of EPO. Cells cultured in the presence of EPO exhibited an elevated apoptotic response to cancer chemotherapeutic agents such as daunorubicin (Dauno) and vinblastine (VBL). Chemosensitization by EPO did not involve an increase in p53 activation, yet correlated with enhanced Bax/Bak-dependent mitochondrial membrane perturbation and caspase maturation. In vitro monotherapy with Dauno or VBL induced the degradation of IkappaBalpha, provoked the translocation of NF-kappaB p65/50 to the nucleus and stimulated the expression of an NF-kappaB-activatable reporter gene. All these signs of NF-kappaB activation were perturbed in the presence of EPO. Inhibition of JAK2, one of the receptor-proximal elements of EPO-mediated signal transduction, greatly diminished the EPO-mediated chemosensitization and NF-kappaB inhibition. EPO lost its death-facilitating effects in the presence of an NF-kappaB inhibitor, underscoring the cause-effect relationship between EPO-mediated chemosensitization and NF-kappaB inhibition. Altogether, these results suggest that, at least in a specific subset of tumors, EPO receptor agonists can prevent activation of the NF-kappaB pathway, thereby enhancing the propensity of EPO receptor-positive tumor cells to undergo apoptosis.

Topics: Antineoplastic Agents; Apoptosis; Carcinoma, Renal Cell; Cell Death; Cell Line, Tumor; Daunorubicin; Erythropoietin; Humans; Janus Kinase 2; Kidney Neoplasms; NF-kappa B; Protein Transport; Protein-Tyrosine Kinases; Proto-Oncogene Proteins; Receptors, Erythropoietin; Signal Transduction; U937 Cells; Vinblastine

Von Hippel-Lindau (VHL) disease is characterized by multiple tumors in specific target organs. The tumors at different sites share distinct morphologic and genetic characteristics but their cell of origin is unknown. We show that VHL disease-associated renal clear cell carcinomas (RCC) consistently coexpress erythropoietin (Epo) and Epo receptor (EpoR). In addition, coexpression of Epo and EpoR is detected in many renal cysts, providing further evidence that renal cysts are potential precursors for RCC. In conjunction with VHL gene deficiency, coexpression of Epo and EpoR in renal cysts and tumors may reflect a developmental arrest in immature mesenchymal cells. Such arrest may lead to autocrine stimulation, cell proliferation, and renal tumor development, similar to tumorigenesis of VHL disease-associated hemangioblastomas.

Topics: Blotting, Western; Carcinoma, Renal Cell; Erythropoietin; Female; Gene Expression; Humans; Immunohistochemistry; Kidney Diseases, Cystic; Kidney Neoplasms; Male; Receptors, Erythropoietin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm; von Hippel-Lindau Disease

Erythropoietin (EPO)-producing renal cell carcinomas (RCC) in patients with chronic renal failure secondary to autosomal dominant polycystic kidney disease (ADPKD) has not previously been reported. We report a case of EPO-producing RCC associated with ADPKD in a 66-year-old woman, and discuss the clinical and radiological findings.

Topics: Aged; Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Kidney Failure, Chronic; Kidney Neoplasms; Neoplasm Proteins; Polycystic Kidney, Autosomal Dominant

An obese 76-year-old woman with type II diabetes, hypertension, coronary artery disease, and gastroesophageal reflux was found to have a 6-cm lower-pole mass in a solitary functional right kidney. Because her religious beliefs prohibited blood transfusion, minimally invasive surgery--a laparoscopic partial nephrectomy--was performed, with a good result. Minimally invasive surgery, perhaps with administration of erythropoietin, iron-dextran, or both, is often a good option for severely anemic patients or those whose religious beliefs are opposed to transfusion. Methods of minimizing blood loss intraoperatively are reviewed.

Topics: Aged; Carcinoma, Renal Cell; Coronary Disease; Diabetes Mellitus, Type 2; Erythropoietin; Female; Gastroesophageal Reflux; Humans; Hypertension; Iron-Dextran Complex; Jehovah's Witnesses; Kidney Neoplasms; Laparoscopy; Nephrectomy

Erythropoietin (EPO)-producing renal cell carcinomas in two hemodialysis patients are reported. Despite deteriorated kidney function, these patients did not manifest anemia at diagnosis and their elevated serum EPO levels rapidly returned to within the normal range after nephrectomy. Immunohistochemical staining of the resected specimens showed production of erythropoietin in the tumor cells in one case and in the lining cells of the cyst wall in the other case. Renal cell carcinoma could cause an increase of blood hematocrit level in dialysis patients.

Topics: Carcinoma, Renal Cell; Erythropoietin; Humans; Immunohistochemistry; Kidney Neoplasms; Male; Middle Aged; Nephrectomy; Renal Dialysis

Topics: Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Kidney Neoplasms; Middle Aged; Risk Factors

An increased level of soluble transferrin receptor (sTfR) has been recognized as a useful indicator of iron deficiency, especially in tumour anaemia and in chronic diseases. In cases of erythropoietin substitution, however, it indicates a successful stimulation of erythropoiesis. We report an "unusual" increase in sTfR in a 60-year-old man who suffered from end-stage hypernephroma with extended lung metastases. He showed pulmonal hypertension, polycythaemia and a high serum level of erythropoietin. We assume that, in this case, the increased sTfR originates not only from bone marrow but could be partly contributed also by the malignant tissue of the hypernephroma.

Topics: Anemia, Iron-Deficiency; Biopsy; Bone Marrow; Carcinoma, Renal Cell; Diagnosis, Differential; Erythropoietin; Humans; Kidney Neoplasms; Male; Middle Aged; Neoplasm Proteins; Polycythemia; Receptors, Transferrin

Topics: Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Neoplasms; Ligases; Oxygen; Transcription Factors; Tumor Suppressor Proteins; Ubiquitin-Protein Ligases; von Hippel-Lindau Disease; Von Hippel-Lindau Tumor Suppressor Protein

The von Hippel-Lindau (VHL) tumor suppressor gene targets hypoxia-inducible transcription factors (HIFs) for proteasomal degradation. Erythrocytosis due to inappropriate production of erythropoietin (EPO), one of the HIF target genes, is a classic albeit rare finding in patients with renal cancer. We report the clinical to molecular analysis in a patient in whom a thrombotic myocardial infarction was the first manifestation of a clear cell renal carcinoma associated with an elevated serum EPO level (109 U/L) and erythrocytosis (hemoglobin 200 g/L [20 g/dL]). The tumor strongly expressed EPO messenger RNA and the 2 regulatory subunits HIF-1alpha and HIF-2alpha. Sequence analysis of tumor tissue identified a point mutation of the VHL gene (nucleotide 701 T > C) with a predicted amino acid exchange (Leu163Pro). This structural change, although located at distance to the HIF-binding region, was found to inhibit binding of HIF-1alpha to VHL, thus leading to accumulation of HIF, which drives EPO production.

Topics: Carcinoma, Renal Cell; Erythropoietin; Genes, Tumor Suppressor; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Kidney Neoplasms; Ligases; Male; Middle Aged; Myocardial Infarction; Point Mutation; Polycythemia; RNA, Messenger; Tomography, X-Ray Computed; Transcription Factors; Tumor Cells, Cultured; Tumor Suppressor Proteins; Ubiquitin-Protein Ligases; Von Hippel-Lindau Tumor Suppressor Protein

To analyze the secretion of hematopoietic growth factors and the expression of their corresponding receptors in 40 newly established renal cell carcinoma (RCC) cell lines of different histologic types. Little is known about the secretion and function of hematopoietic growth factors by human RCCs.. The expression of the hematopoietic growth factors (ie, erythropoietin, interleukin [IL]-3, IL-5, granulocyte colony-stimulating factor [G-CSF], granulocyte-macrophage colony-stimulating factor [GM-CSF], and macrophage colony-stimulating factor [M-CSF]) was determined by enzyme-linked immunosorbent assay analysis under different culture conditions, including suspension culture and monolayer cultures (plastic and Matrigel-coated culture flasks). The expression of their corresponding receptors was defined by fluorescence activated cell scanner analysis and by reverse-transcriptase polymerase chain reaction. The response of the RCC cell lines to exogenous hematopoietic growth factors was analyzed by MTT assay.. In almost all of the cell lines, significant amounts of GM-CSF and M-CSF were secreted, and in four cell lines, a secretion of G-CSF was detected. Fourteen RCC cell lines showed secretion of IL-3, and production of IL-5 and erythropoietin was not observed in any cell line. Secretion of GM-CSF and M-CSF was affected by the substratum offered for cell attachment in the adherent cultures. GM-CSF secretion was more pronounced under culture conditions with a reduced frequency of cell-to-cell contacts. Two cell lines were shown to express receptors for M-CSF, but receptors for G-CSF and GM-CSF could not be detected in any cell line. Exposure to exogenous G-CSF, GM-CSF, and M-CSF did not affect the proliferation of our RCC cell lines.. The results of our study clearly demonstrate that human RCC cells can secrete significant amounts of G-CSF, GM-CSF, M-CSF, and IL-3, and are thereby theoretically able to modulate the host's tumor-directed immune response.

Topics: Carcinoma, Renal Cell; Enzyme-Linked Immunosorbent Assay; Erythropoietin; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoietic Cell Growth Factors; Humans; Interleukin-3; Interleukin-5; Kidney Neoplasms; Macrophage Colony-Stimulating Factor; Neoplasm Proteins; Tumor Cells, Cultured

We reported recently that normal human, rat, and mouse tubular cells express authentic erythropoietin-receptors (EPO-R) through which EPO stimulates mitogenesis. The present study examines whether EPO could elicit such a proliferative and thereby potentially detrimental response in cells of human renal-cell carcinoma (RCC).. Nephrectomy samples were screened from patients with RCC (one chromophilic, two clear cell) as well as cell lines of human (Caki-2, 786-0) and mouse (RAG) renal adenocarcinomas for expression of EPO-R transcripts and protein. Cells were further tested for specific 125I-EPO binding and mitogenic response to EPO.. Authentic EPO-R transcripts and protein (approximately 72 kD) were detected in renal tumors and cell lines. Tumors showed low-level EPO expression, while cell lines did not. In cells, specific 125I-EPO binding to a single class of EPO-R (apparent Kd 1. 3 to 1.4 nmol/L, Bmax 2.2 to 2.6 fmol/mg protein) was observed. EPO stimulated cell proliferation dose dependently, and the individual mitogenic effects of either EPO or 10% newborn calf serum were markedly amplified when both were coadministered.. These data are the first to demonstrate, to our knowledge, that human RCCs express EPO-R message and protein and that receptor activation stimulates their proliferation in vitro. If these mitogenic effects of EPO are also operative in patients with RCC, endogenous EPO or its administration for the treatment of anemia could potentially hasten proliferation of renocellular malignancies.

Topics: Adenocarcinoma, Clear Cell; Anemia; Animals; Carcinoma, Renal Cell; Cell Division; Erythropoietin; Gene Expression; Humans; Iodine Radioisotopes; Kidney Neoplasms; Kidney Tubules, Proximal; Male; Membrane Proteins; Mice; Mice, Inbred BALB C; Middle Aged; Mitogens; Neovascularization, Pathologic; Receptors, Erythropoietin; RNA, Messenger; Transcription, Genetic; Tumor Cells, Cultured; von Hippel-Lindau Disease

A 58 year old male heavy smoker presented with intracranial haemorrhage and erythrocytosis. Four aetiologies of polycythaemia--polycythaemia rubra vera (PRV), renal cell carcinoma, sleep apnoea syndrome, and relative polycythaemia--were found to be associated with the underlying causes of erythrocytosis. He did not fulfill the diagnostic criteria for PRV at initial presentation, but an erythropoietin independent erythroid progenitor assay identified the masked PRV, and the low post-phlebotomy erythropoietin concentration also suggested the likelihood of PRV evolution. This case demonstrates that a search for all the possible causes of erythrocytosis is warranted in patients who already have one aetiology of polycythaemia.

Topics: Carcinoma, Renal Cell; Erythroid Precursor Cells; Erythropoietin; Humans; Intracranial Hemorrhages; Kidney Neoplasms; Male; Middle Aged; Polycythemia; Polycythemia Vera; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sleep Apnea Syndromes; Smoking

We assessed blood loss and subsequent transfusion associated with nephrectomy performed for suspected renal cell carcinoma to establish guidelines for preoperative autologous blood donation and identify a subgroup of patients that may benefit from erythropoietin administration.. We retrospectively reviewed the charts of 211 patients who underwent partial (73%) or radical (23%) nephrectomy for presumed renal cell carcinoma at our institution between 1990 and 1999. Patients were divided into groups 1-44.5% treated with radical nephrectomy for localized disease, 2-21.3% radical nephrectomy for metastatic lesions invading the renal vasculature or inferior vena cava, 3-8% radical nephrectomy for metastatic disease with locally extensive lesions and 4-26.5% partial nephrectomy for localized lesions. Patient charts were evaluated for preoperative and postoperative hematocrit, estimated blood loss, transfusions received, surgical complications and underlying disease.. Median estimated blood loss was 200, 400, 250 and 555 cc in groups 1 to 4, respectively. However, patients in groups 2 and 3 had a substantially greater range of blood loss than those in groups 1 and 4, respectively. The incidence of those with a blood loss of greater than 1 l. was 7%, 36%, 24% and 11% in groups 1, to 4, respectively. The incidence of those requiring transfusion was significantly lower in group 1 than in groups 2 to 4 (18% versus 44%, 24% and 30%, respectively, p <0.009). Mean transfusion requirement plus or minus standard deviation was significantly greater in groups 2 and 3 than in 1 and 4 (2.3 +/- 1.08, 5.5 +/- 4.4, 11.3 +/- 9.6 and 2.3 +/- 1.7 units, respectively, p <0.05). No significant difference was noted in the change in hematocrit as a result of surgery in the 4 groups (p >0.05). Similarly underlying disease and operative complications did not have a significant effect on blood loss or transfusion (p >0. 05).. Radical or partial nephrectomy for localized renal cell carcinoma leads to consistent and well tolerated operative blood loss that rarely results in the need for substantial transfusion. In contrast, nephrectomy for advanced disease may cause a risk of greater blood loss and subsequent need for the transfusion of multiple units of blood. While preoperative autologous blood donation may have limited value in this regard due to the high cost and number of units needed, preoperative erythropoietin administration may be a viable option. Prospective randomized studies are currently planned.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Blood Loss, Surgical; Blood Transfusion; Carcinoma, Renal Cell; Erythropoietin; Hematocrit; Humans; Kidney Neoplasms; Middle Aged; Nephrectomy; Recombinant Proteins; Retrospective Studies

A 57-year-old man with renal cell carcinoma and erythrocytosis showed a high serum level of erythropoietin (EPO). High EPO signal was observed on Northern blot analysis and RT-PCR in the total RNA extracted from the renal tumor. Immunohistochemical staining also demonstrated tumor tissue with high immunostaining of EPO. Nucleotide sequences of EPO cDNA in the tumor were normal. To date, only one report has discussed the nucleotide sequences of a tumor's EPO gene; it showed mutant EPO cDNA in hepatocellular carcinoma tissue. This is the first demonstration of normal EPO cDNA in renal cell carcinoma.

Topics: Carcinoma, Renal Cell; DNA, Complementary; Erythropoietin; Gene Expression Regulation, Neoplastic; Humans; Kidney Neoplasms; Male; Middle Aged

Erythropoietin (EPO) is a well-recognized hormone that induces erythrocytosis in a wide range of physiologic and pathologic situations in mammals. One of these situations is a paraneoplastic erythrocytosis, which might be seen in association with various neoplasms, including renal cell carcinoma (RCC), hepatoma, and cerebellar hemangioblastoma. Although there have been multiple studies confirming the association between this erythrocytosis and the production of EPO by tumor cells, immunohistochemical detection of EPO in formalin-fixed, paraffin-embedded tissue was not described. We report on the use of microwave antigen retrieval to detect EPO in RCCs in such routinely processed tissues. We selected 19 RCCs received as nephrectomy specimens, fixed in formalin, and routinely processed. These cases were previously diagnosed on the basis of morphologic, immunohistochemical, and clinical features. We examined the immunoreactivity of these specimens with a monoclonal anti-EPO antibody. Fetal (20 wk gestational age) liver served as a positive control. Intense positive immunoreactivity was observed as cytoplasmic, granular staining in fetal hepatocytes. Fourteen RCCs (10 clear cell and 4 tubulopapillary types) demonstrated unequivocal cytoplasmic immunoreactivity. Additionally, two clear cell tumors were only focally positive, whereas three (16%) were negative. EPO immunoreactivity might thus prove to be of value in the diagnosis or confirmation of RCC, particularly in the context of routinely processed material.

Topics: Antibodies, Monoclonal; Carcinoma, Renal Cell; Erythropoietin; Histocytological Preparation Techniques; Humans; Immunohistochemistry

A 49-year-old woman had been on hemodialysis for 18 years. She presented with left back pain and macrohematuria. Radiologic studies demonstrated a left renal tumor with acquired cystic disease of the kidney. Her serum erythropoietin (EPO) level was 78.4 U/L despite no history of EPO supplementation. Left radical nephrectomy was performed. Pathologic examination revealed EPO-producing renal cell carcinoma. After surgery, the patient's serum EPO level decreased markedly to 15.1 U/L. The measurement of serum EPO levels may be useful for detecting and monitoring a recurrence of renal cell carcinoma with acquired cystic disease of the kidney in patients on long-term hemodialysis.

Topics: Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Immunohistochemistry; Kidney Neoplasms; Middle Aged; Nephrectomy; Polycystic Kidney Diseases; Tomography, X-Ray Computed

Topics: Carcinoma, Renal Cell; Circadian Rhythm; Erythropoietin; Humans; Kidney; Kidney Neoplasms; Male; Middle Aged

A case of erythropoietin-producing renal cell carcinoma is reported. A 59-year-old woman was referred to our hospital for further examination of erythrocytosis accompanied with an elevated serum erythropoietin level. Ultrasonography and computerized tomography scan showed a left renal tumor. She underwent a left radical nephrectomy. Postoperatively, serum erythropoietin level and erythrocytosis were normalized. Histopathology was renal cell carcinoma, clear cell subtype, and immunohistochemistry revealed erythropoietin in cancer cells. Furthermore, the cancerous tissue contained a high concentration of erythropoietin.

Topics: Biomarkers, Tumor; Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Immunohistochemistry; Kidney Neoplasms; Middle Aged; Polycythemia

A cell line obtained from a primary lesion of renal cell carcinoma was newly established and observed to produce erythropoietin (Ep) continuously in vivo as well as in vitro. The histopathological and biological characteristics of this cell line were then analyzed in parallel with a study initiated on the role of cAMP on Ep production.. The cells were subsequently serially transplanted into nude mice. The levels of Ep and hematocrit in the mice were found to correlate very closely with the tumor size. The histology of the xenograft was similar to the original tumor cells which constituted a nearly clear cell type and was immunostained positively for Ep. Ultrastructurally, Ep was localized in the perinuclear space, with part of the rough surface of the endoplasmic reticulum. In vitro, the cells grew exponentially with an approximate population doubling time of 2 days. The mRNA of Ep was detected using the RT-PCR method. The number of chromosomes in the cells ranged from 117 to 147 and featured complicated rearrangements and marker chromosomes. Forskolin, a well-known activator of adenylate cyclase which in turn generated the accumulation of cyclic AMP, produced a significant dose-related enhancement of Ep biosynthesis both in the cells and in the spent culture medium.. Based on the results derived from the foregoing analysis of this new cell line, cAMP was found to play a salient role both in Ep biosynthesis as well as in Ep release.

Topics: Aged; Animals; Carcinoma, Renal Cell; Cyclic AMP; Erythropoietin; Humans; Kidney Neoplasms; Male; Mice; Tumor Cells, Cultured

Erythropoietin production by renal cell carcinoma (RCC) is reported to be a potential marker for interleukin-2/interferon-alpha-responding tumor. We have investigated whether erythropoietin of RCC cells is involved in the immune recognition by lymphokine-activated killer (LAK) cells. Cells from primary culture of RCC cells expressing erythropoietin-mRNA or producing erythropoietin were more susceptible to lysis by LAK cells than those not expressing or producing it, respectively. RCC cells transfected with erythropoietin-cDNA became more susceptible to lysis by LAK cells than their erythropoietin-negative parental cells. These results indicate higher susceptibility of erythropoietin-producing RCC cells to lysis by LAK cells, suggesting that erythropoietin of RCC cells is involved in the immune recognition by LAK cells.

Topics: Carcinoma, Renal Cell; Cytotoxicity, Immunologic; DNA, Complementary; Erythropoietin; Humans; Kidney Neoplasms; Killer Cells, Lymphokine-Activated; Transfection; Tumor Cells, Cultured

Topics: Adrenal Gland Neoplasms; Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Recombinant Proteins

Topics: Adult; Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Neoplasms; Male; Neoplasm Invasiveness; Recombinant Proteins; Treatment Outcome

We conducted the establishment of erythropoietin (Epo) producing human renal cell carcinoma heterotransplanted in nude mice (JRC 901) and analysed its histopathological and biological characteristics. Regarding to histopathological analysis, JRC 901 showed renal cell carcinoma with granular cell subtype, alveolar pattern and grade II malignancy. In an effort to the electron microscopic analysis, JRC 901 showed renal cell carcinoma with microvilli, rich lipid droplets and mitochondria. As to the tumour doubling time, the JRC 901 showed 14.81 days in a logarithmic phase. As to the karyotype, the JRC 901 showed human, 46, XY, -11, 8p+, 17q-, +mar. After tumour inoculation to the nude mice, the blood level of Epo increased at 5 weeks, and its level reached at 485.2 mU/ml at 12 weeks after tumour inoculation. Furthermore, immunohistochemical staining using anti-Epo showed positive staining within cytoplasm of JRC 901. Moreover, the production of Epo was observed in the level of mRNA (264 bp) using RT-PCR method. We conclude that the JRC 901 is a human renal cell carcinoma heterotransplantable to nude mice and this tumour produce the Epo after tumour inoculation to nude mice.

Topics: Animals; Carcinoma, Renal Cell; Cell Line; Erythropoietin; Humans; Immunohistochemistry; Kidney Neoplasms; Male; Mice; Mice, Nude; Middle Aged; Neoplasm Transplantation; Transplantation, Heterologous; Tumor Cells, Cultured

The paraneoplastic syndrome of erythrocytosis is associated with a variety of neoplasms including renal adenocarcinoma, cerebellar hemangioma, and hepatoma. We now report the characterization of the biological and molecular features of an erythropoietin-secreting human renal adenocarcinoma, designated RCCEp+. Serial transplantation of the tumor in athymic mice resulted in a dramatic increase in hematocrit and serum erythropoietin concentration. Growth in vitro was accompanied by a constant rate of erythropoietin secretion. Karyotype analysis demonstrated several unusual features, including the absence of 3p deletions and near tetraploidy. Erythropoietin mRNA was demonstrated by Northern blot both in freshly excised tumor and in tumor cells growing in vitro. Erythropoietin secretion was constitutive and was not induced either by cobalt or hypoxia. Southern blot analysis revealed no rearrangement of the erythropoietin gene in the tumor. Interestingly, in situ hybridization demonstrated erythropoietin mRNA in only a small population of the tumor cells. Further studies of RCCEp+ should prove useful in elucidating the molecular basis for this paraneoplastic syndrome.

Topics: Animals; Carcinoma, Renal Cell; Erythropoietin; Female; Gene Expression; Humans; In Situ Hybridization; In Vitro Techniques; Kidney Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Paraneoplastic Syndromes; Polycythemia; RNA, Messenger; Transplantation, Heterologous; Tumor Cells, Cultured

The prevalence of increased serum immunoreactive erythropoietin (Epo) was determined in a prospective study of 49 patients with renal cell carcinoma. Measured by a monoclonal antibody based commercial enzyme-linked immunoassay, the Epo concentration was above the normal range, determined in nonanemic humans, in four of the renal carcinoma patients. Since three of these were anemic, their increased Epo level was considered to be appropriate. The high estimate of serum Epo (218 U/l) in the fourth patient, who was not anemic, was not confirmed when tested by radioimmunoassay. Thus, in contrast with earlier studies, our results indicate that increased Epo is not a clear serological renal cell carcinoma marker. In addition, when monolayer cell cultures of 14 different established human renal carcinoma lines were screened, none of these released immunoreactive Epo in measurable amounts.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Renal Cell; Cell Hypoxia; Enzyme-Linked Immunosorbent Assay; Erythropoietin; Female; Humans; Kidney Neoplasms; Male; Middle Aged; Paraneoplastic Syndromes; Prevalence; Prospective Studies; Tumor Cells, Cultured

An 85-year-old woman had a right-sided renal cell carcinoma removed 20 years ago. At haemoglobin concentration. Two years ago she had a syncope, at which time the haemoglobin concentration was 16.9 g/dl. Ultrasound and computed tomography (CT) revealed an extensive retroperitoneal space-occupying lesion, which however was not investigated further, and no therapeutic consequences were drawn. An erythrocytosis (7.5 x 10(6)/microliters) and elevated haemoglobin concentration (> 20 g/dl) were found when she was examined after a fall in which she had sustained only minimal injury. The retroperitoneal mass had slightly increased in size. Histological examination of a CT-guided fine-needle biopsy revealed metastases of the hypernephroid carcinoma. The serum erythropoietin concentration was increased (42.4 U/l) and failed to increase even after repeated venesections, indicating erythropoietin production by the late metastases of the renal cell carcinoma. There was no evidence for any systemic haematological disease. Six months after the diagnosis of metastases the patient died at home, presumably of a cerebrovascular accident.

Topics: Aged; Aged, 80 and over; Carcinoma, Renal Cell; Erythropoietin; Fatal Outcome; Female; Humans; Kidney Neoplasms; Polycythemia; Retroperitoneal Neoplasms; Time Factors

The discovery of high hemoglobin and hematocrit values in a patient necessitates the determination of the red blood cell mass in order to confirm the absolute character of the polycythemia. If a true polycythemia is confirmed, its etiology must then be established. The diagnostic approach of polycythemia is presented in this paper. It is illustrated by a case presentation in which a polycythemia secondary to a renal carcinoma is discussed. Erythrocytosis is a classical, albeit rare manifestation of this type of tumor, and has the advantage of allowing early detection. Thus, it permits a prompt treatment plan, thereby improving the prognosis of such a neoplasia. The usefulness of a serum level of erythropoietin (EPO) is subsequently discussed. The diagnostic value of EPO remains controversial because of the overlapping values recorded amongst healthy patients, patients with polycythemia vera and others with secondary polycythemia. Finally, we discuss the presence of substances in paraneoplastic polycythemias whose biological activity is close to that of EPO. However, this molecules of a different structure would not be detected by the radioimmunoassay used to measure erythropoietin level.

Topics: Algorithms; Carcinoma, Renal Cell; Diagnosis, Differential; Erythropoietin; Humans; Kidney Neoplasms; Male; Polycythemia

Interleukin-2 (IL-2) recently was approved by the Food and Drug Administration for the treatment of renal cell cancer. It is effective in a small minority of patients, but no markers identify individuals likely to respond to treatment.. Two polycythemic patients with erythropoietin-producing renal cell cancer and three other polycythemic patients with renal cell cancer were treated with the combination of IL-2 and alpha-interferon (alpha-IFN).. All five patients achieved a partial or complete remission. In both patients in which it was measured, the erythropoietin level decreased significantly with treatment, and the polycythemia resolved in all patients. Hypothyroidism developed in two patients, and transient hyperthyroidism developed in another.. These results contrast with those achieved with IL-2 alone or in combination with lymphokine-activated killer cells, for which a 15% response rate was seen in patients with renal cell cancer and polycythemia. Although less than 5% of renal cell tumors produce erythropoietin, its production may identify a subset of individuals with renal cell cancer responsive to IL-2 and alpha-IFN.

Topics: Adult; Aged; Biomarkers; Carcinoma, Renal Cell; Drug Therapy, Combination; Erythropoietin; Female; Humans; Interferon-alpha; Interleukin-2; Kidney Neoplasms; Male; Middle Aged; Polycythemia; Remission Induction

Erythropoietin levels in serum were analyzed in 165 patients with renal cell carcinoma. All samples were taken before therapy and stored at -80 degrees C. Erythropoietin, a glucoprotein produced by the renal cortex was quantified by an enzyme immunoassay. Fifty-five of 165 patients (33%) had elevated serum levels. In patients with metastatic disease (M+), elevated levels were found in 24 of 65 cases (38%). Patients with high-grade tumors had significantly more often increased erythropoietin than those with low-grade tumors. No correlation between erythrocytosis and elevation of erythropoietin in serum was found. There was a significant difference in survival between patients with normal and patients with elevated erythropoietin levels (p = 0.013). The study shows that erythropoietin is a tumor marker with a low sensitivity. However, it correlates with stage and grade and provides prognostic information.

Topics: Biomarkers, Tumor; Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Kidney; Kidney Neoplasms; Male; Middle Aged; Neoplasm Staging; Prognosis; Sensitivity and Specificity; Survival Analysis

An 83-year-old man came to our hospital complaining of asymptomatic gross hematuria. We found a subcutaneous tumor in his chest and an abdominal mass in his left upper quadrant. Laboratory data showed red blood cell count of 737 x 10(4)/mm3 and hemoglobin level of 17.9 g/dl. The serum erythropoietin level measured by radioimmunoassay was considerably high. He underwent left nephrectomy and tumor resection in the chest wall. Serum erythropoietin level became normal following surgery and erythrocytosis disappeared. Histological findings revealed renal cell carcinoma, alveolar type, clear cell subtype, grade 2. The erythropoietin levels of the extracts of the cancer tissue and the normal kidney tissue were 2430 mU/g and 59.5 mU/g, respectively. Lung, liver and bone metastases appeared four months after the operation and serum erythropoietin level increased again.

Topics: Aged; Aged, 80 and over; Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Neoplasms; Male; Skin Neoplasms

Immunotherapy with interleukin-2 (IL-2) and lymphokine-activated killer (LAK) cells results in significant tumor regression in patients with advanced cancer. We have investigated the kinetics of circulating erythroid (BFU-E) and granulocytic-macrophage (CFU-GM) progenitors after IL-2 therapy in 11 cancer patients, mainly affected by metastatic melanoma and renal cell carcinoma. Administration of IL-2 from day 1 through day 5 constantly induced a dramatic decrease of the number of circulating BFU-E and CFU-GM, which then showed a striking rebound (up to values fourfold and sevenfold higher, respectively, than the pretherapy levels) on discontinuation of IL-2, ie, from day 5 through day 10. A similar kinetic pattern was observed during and after the second cycle of IL-2 administration. 3[H]-thymidine killing experiments showed that the cycling activity of the progenitors was virtually unmodified in the rebound phases. To explore the mechanism(s) underlying this kinetic pattern, we have analyzed the plasma concentration of several hematopoietic growth factors, including IL-1 beta, IL-3, IL-4, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), G-CSF, and erythropoietin (Ep). No modifications in the levels of IL-3, GM-CSF, or IL-1 beta were observed, whereas a pronounced increase of IL-6 and G-CSF concentration was monitored, starting at day 3 and peaking at day 5 of treatment (a parallel, but modest, increase of Ep level was also observed). The elevation of IL-6 and G-CSF concentration is directly correlated with and may, at least in part, underlie the subsequent rebound of circulating hematopoietic progenitors. Furthermore, the increase in IL-4 level observed at day 10 of therapy may mediate the eosinophilia gradually starting at this stage of treatment.

Topics: Adult; Aged; Carcinoma, Renal Cell; Cell Count; Erythroid Precursor Cells; Erythropoietin; Female; Granulocyte Colony-Stimulating Factor; Granulocytes; Growth Substances; Hematopoietic Stem Cells; Humans; Immunotherapy, Adoptive; Interleukin-2; Interleukin-6; Kidney Neoplasms; Kinetics; Macrophages; Male; Melanoma; Middle Aged; Neoplasms

The present studies were undertaken to assess the effects of atrial natriuretic factor (ANF) on erythropoietin (Ep) secretion in Ep-producing renal carcinoma (RC) cells using a sensitive radioimmunoassay for Ep. Human ANF produced a significant dose-related increase in Ep secretion at concentrations of 10(-7) and 10(-6) M when compared with vehicle controls. ANF (greater than or equal to 10(-9) M) also significantly increased the intracellular guanosine 3',5'-cyclic monophosphate (cGMP) concentration after 5-min incubation with the RC cells. Scatchard analysis of the human 125I-labeled ANF binding data indicated that the RC cells contain a single class of binding sites with a dissociation constant (Kd) of 93 +/- 1 pM and a binding capacity of 2,190 +/- 750 sites/cell. Incubation of the RC cells with 8-bromo-cGMP in concentrations of 10(-7)-10(-5) M also produced a significant dose-related enhancement of Ep secretion. These findings suggest that the increase in Ep secretion in response to ANF can be attributed, at least in part, to activation of guanylate cyclase, which is coupled to specific ANF receptors on the RC cell.

Topics: Atrial Natriuretic Factor; Carcinoma, Renal Cell; Cyclic GMP; Erythropoietin; Humans; Kidney Neoplasms; Kinetics; Lung Neoplasms; Tumor Cells, Cultured

One to five percent of human renal cell carcinomas are associated with polycythemia. It is generally assumed that polycythemia results from the secretion of erythropoietin (Epo) by the malignant cells. However, there is no direct proof supporting this hypothesis. Three patients with typical renal adenocarcinoma and polycythemia were studied. All three exhibited high Epo serum levels as measured by radioimmunoassay (RIA). A strong Epo signal was observed on Northern blot analysis of total RNA extracted from the renal tumors. The Epo message seemed to be of normal size and no Epo gene rearrangement was observed with the restriction enzymes tested. Using the in situ hybridization technique, a significant labeling was constantly observed on the tumor cells. Immunohistochemical studies showed that these tumor cells, known to be of tubular origin, were labeled by an anti-cytokeratin antibody and therefore were of epithelial nature. Thus, this study demonstrated that malignant cells of tubular origin were able to produce Epo constitutively, whereas in the mouse hypoxic kidney, peritubular cells (probably capillary endothelial cells) were the major site of Epo synthesis.

Topics: Adenocarcinoma; Blotting, Southern; Carcinoma, Renal Cell; DNA Probes; DNA, Neoplasm; Epithelium; Erythropoietin; Gene Rearrangement; Hormones, Ectopic; Humans; Kidney Neoplasms; Nucleic Acid Hybridization; Polycythemia; RNA, Messenger; RNA, Neoplasm

A case of renal cell carcinoma with erythrocytosis is reported. A 58-year-old male had a right renal tumor with retroperitoneal lymph node and lung metastases. Preoperative laboratory data revealed a red blood cell count of 6.31 million/mm3, a hemoglobin concentration of 17.2 g/dl, a hematocrit of 52.1, and an erythropoietin (EPO) level of 47.3 mU/ml, which was determined by RIA method (normal range: 8-30). The patient underwent radical nephrectomy and retroperitoneal lymph node dissection without any blood transfusion because blood loss was only 300 g. The removed kidney weighed 400 g. Pathological examination showed a renal cell carcinoma not invading the fibrous capsule with lymph node metastasis. The EPO activities of extracts from the normal kidney and cancer tissues were less than 18 and 50.8 mU/g, respectively. At present, 3 months postoperatively, he receives daily UFT and HLBI every other day for the lung disease left, which does not change in size, and all laboratory data mentioned above have returned to normal ranges. These results enabled us to diagnose the present case as EPO-producing renal cell carcinoma.

Topics: Carcinoma, Renal Cell; Erythropoietin; Humans; Kidney Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Polycythemia

We used a battery of antigens to determine whether immunohistochemistry can (a) contribute to resolving the histogenesis of the stromal component of the capillary hemangioblastoma, and (b) answer cases of difficult pathologic differential diagnosis with metastatic clear cell carcinoma. The stromal cells of the capillary hemangioblastoma are antigenically polymorphous and may express immunoreactive erythropoietin, renin, keratin, Leu M1, Leu 7, actin, neuron-specific enolase, S100 protein, and glial fibrillary acidic protein. However, the use of epithelial membrane antigen allows certain histopathologic distinction between capillary hemangioblastoma and metastatic clear cell carcinoma.

Topics: Adenocarcinoma; Aged; Antigens, Differentiation; Antigens, Neoplasm; Carcinoma, Renal Cell; Cerebellar Neoplasms; Diagnosis, Differential; Erythropoietin; Hemangiosarcoma; Humans; Immunoenzyme Techniques; Keratins; Kidney Neoplasms; Male; Membrane Glycoproteins; Mucin-1; Renin

Erythropoietin producing cells were identified in the murine hypoxic kidney by in situ hybridization. The positive cells were peritubular cells, most likely endothelial cells of the cortex and outer medulla. Glomerular and tubular cells were not labelled. In three patients with renal adenocarcinomas associated with polycythemia, a strong Epo message was observed on Northern blot analysis. Using in situ hybridization, a strong labelling was observed in all cases on the tumor cells which are of tubular origin.

Topics: Animals; Carcinoma, Renal Cell; Erythropoietin; Gene Expression Regulation; Genes; Humans; Hypoxia; Kidney Cortex; Kidney Neoplasms; Mice; Neoplasm Proteins; Nucleic Acid Hybridization; Polycythemia; Transcription, Genetic

We have presented a model for the role of external messenger substances in hypoxic stimulation of kidney production of erythropoietin. These autacoids probably act in concert to activate the adenylate cyclase system to enhance production and/or secretion of erythropoietin. The phosphoproteins generated in this system could act at the level of transcription and translation of erythropoietin as well as at the level of release of erythropoietin from the cell. Even though eicosanoids and beta-2-adrenergic agonists may be involved in mild to moderate hypoxia, it seems more likely that adenosine is more involved in erythropoietin production with increasing severity of hypoxia. Adenosine may play a very early role in hypoxia following the decrease in ATP to trigger erythropoietin production, and hydrogen peroxide may be generated from hypoxanthine, a metabolite of adenosine, during reoxygenation and regional changes in blood flow in the normal kidney and perhaps in certain renal and hepatic tumors. Further work is necessary in vivo to completely clarify the role of adenosine and oxygen free radicals in regulating kidney production of erythropoietin.

Topics: Adenosine; Animals; Carcinoma, Renal Cell; Catalase; Erythropoietin; Female; Glucose Oxidase; Humans; Hydrogen Peroxide; Hypoxanthine; Hypoxanthines; Kidney; Kidney Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Superoxides; Tumor Cells, Cultured; Xanthine Oxidase; Xanthines

We have identified a monoclonal antibody (ERY-1), which reacts with erythrocytes, erythroid precursor cells, and with embryonal yolk sac, and normal liver and kidney. The antibody also decorates the neoplastic cells of hepatocellular, renal, and yolk sac carcinomas. No reactivity was seen in a variety of other epithelial or mesenchymal neoplasms. It is possible that ERY-1 recognizes an erythropoiesis-associated antigen present in yolk sac, kidney, liver, and bone marrow, all of which are involved in erythropoiesis in various stages of human development. Furthermore, ERY-1 has proved to be extremely useful in the histopathologic diagnosis of hepatocellular and renal cell carcinomas.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Hepatocellular; Carcinoma, Renal Cell; Enzyme-Linked Immunosorbent Assay; Erythroblasts; Erythropoiesis; Erythropoietin; Humans; Immunohistochemistry; Kidney Neoplasms; Liver Neoplasms

This study provides clear documentation of in vivo biogenesis of erythropoietin (Epo) by a human renal carcinoma. A middle-aged woman with a clear cell renal carcinoma of the left kidney developed severe polycythemia. This polycythemia was accompanied by markedly elevated levels of immunoreactive erythropoietin both in the peripheral venous blood, and in blood derived from the left renal vein during nephrectomy. Exstirpation of the non-invasive renal carcinoma was followed by complete restoration of both hematocrit and erythropoietin plasma concentration to normal levels. The fall in plasma erythropietin concentration immediately after nephrectomy (T/2 less than or equal to 3 hours) was probably a valid representation of plasma erythropoietin metabolism in this patient. Direct evidence of erythropoietin production in individual renal carcinoma cells was provided by immunoperoxidase studies demonstrating focal cytoplasmatic accumulation of immunoreactive erythropoietin in the tumor cells.

Topics: Aged; Carcinoma, Renal Cell; Erythropoietin; Female; Half-Life; Hematocrit; Humans; Immunoenzyme Techniques; Kidney Neoplasms; Microscopy, Electron; Nephrectomy; Polycythemia

Topics: 8-Bromo Cyclic Adenosine Monophosphate; Carcinoma, Renal Cell; Colony-Forming Units Assay; Erythropoiesis; Erythropoietin; Hematocrit; Humans; In Vitro Techniques; Kidney Neoplasms; Radioimmunoassay; Tumor Cells, Cultured

An established cloned human renal carcinoma line RC-1, which has been continuously maintained in culture for several years and which produces erythropoietin, was injected s.c. into BALB/c athymic mice and produced tumors. Tumorigenicity was directly correlated with the number of RC-1 cells inoculated. Tumor cell histology resembled the original patient-derived tumor. Tumor-bearing mice developed hepatosplenomegaly and significant reticulocytosis with elevated hemoglobin and hematocrit values that were proportional to tumor mass. In addition, red cell mass and blood volume of nude mice increased over 100% as compared to control mice or to animals bearing nonrelevant neoplasms. Large amounts of immunoreactive erythropoietin could be extracted from the nude mouse RC-1 tumors. These results indicate that the RC-1 cell line is tumorigenic and produces biologically active erythropoietin in vivo in athymic mouse hosts, thus providing a reproducible model to study ectopic erythropoietin production and its regulation in vivo.

Topics: Animals; Carcinoma, Renal Cell; Erythropoiesis; Erythropoietin; Female; Humans; Kidney Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Polycythemia; Transplantation, Heterologous; Tumor Cells, Cultured

Topics: Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Kidney Neoplasms; Middle Aged; Nephrectomy; Radioimmunoassay

We used our recently reported stable, transformed human renal carcinoma cell line as a model system to study the role of 3',5'-adenosine monophosphate (cAMP) in erythropoietin secretion. The erythropoietin produced by these cells is both biologically active and immunologically cross-reactive with purified native human hormone in our radioimmunoassay. Erythropoietin release by these renal carcinoma cells appears to be stimulated by cAMP as well as by the phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (MIX). The response to cAMP involves a rapid and enhanced release of hormone, which occurred within 30 minutes of exposure of the cells to the effector and continued for at least 4 hours. Intracellular erythropoietin was higher in the control cultures than in the cells treated with cAMP, suggesting that cAMP stimulates the release of a storage pool of hormone. The ability of cAMP and MIX to elicit the release of erythropoietin suggests that a cAMP-mediated mechanism is involved in the release of this hormone.

Topics: 1-Methyl-3-isobutylxanthine; 8-Bromo Cyclic Adenosine Monophosphate; Carcinoma, Renal Cell; Cell Line; Cyclic AMP; Erythropoietin; Humans; Secretory Rate; Time Factors

Topics: Animals; Carcinoma, Renal Cell; Cells, Cultured; Erythropoietin; Humans; Immunoenzyme Techniques; Kidney Neoplasms; Mice; Mice, Nude; Neoplasm Transplantation

A human renal carcinoma from a patient with an erythrocytosis, serially transplanted into athymic nude mice, was grown in primary monolayer cell cultures. After reaching confluency the cultured cells formed multicellular hemicysts (domes) which became more abundant as the cultures approached saturation density. Erythropoietin (Ep) production by this renal carcinoma in culture was only slightly increased at the time of semiconfluency but showed a marked increase in Ep levels in the culture medium after the cultures reached confluency, in parallel with an increase in dome formation. The phorbol ester tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) showed a significant dose-related inhibitory effect on Ep production and dome formation in the renal carcinoma cell cultures, suggesting an important role of protein kinase C, the only known receptor for TPA, in inhibiting the expression of differentiated phenotypes in the renal carcinoma cells. TPA also suppressed Ep secretion over a period of 96 h, indicating a time course of suppression of this differentiated function of the renal carcinoma cells in culture. This hypothesis was further supported by the observation that diacylglycerol, the endogenous activator of protein kinase C, likewise inhibited Ep production and dome formation in the renal carcinoma cell cultures. These studies suggest a role of the inositol-lipid second messenger path and protein kinase C in the regulation of Ep production.

Topics: Carcinoma, Renal Cell; Cell Division; Diglycerides; Erythropoietin; Glycerides; Humans; Protein Kinase C; Tetradecanoylphorbol Acetate; Time Factors; Tumor Cells, Cultured

The bioassay using the polythemic mice demonstrated persistent erythropoietin (Ep) activity in 24 renal carcinoma patients. Eight patients without clinical evidence of renal carcinoma had Ep levels that were slightly higher than those of controls, suggesting the possibility of occult disease. Increased levels of Ep were noted in 5 patients with other genitourinary carcinomas. This selective study reaffirms the value of Ep as a biologic marker in some renal cell cancers, and occasionally in other genitourinary tumors.

Topics: Aged; Animals; Biological Assay; Carcinoma, Renal Cell; Erythropoietin; Female; Humans; Kidney Neoplasms; Male; Mice; Mice, Inbred ICR; Middle Aged; Polycythemia; Urogenital Neoplasms

The present studies report the maintenance of erythropoietin (Ep) production in long-term cultures of a human renal carcinoma from a patient with erythrocytosis. The renal carcinoma cells were grown and maintained in monolayer cultures for 7 months. They were serially passaged every 2-3 weeks when the cultured cells reached confluency. Ep levels measured with a sensitive radioimmunoassay in the spent culture media of the cells in the stage of semiconfluent or confluent density were less than 20 and 30 mU/ml, respectively, throughout the period of 15 successive passages. However, when the renal carcinoma cells were maintained in culture without passage after reaching confluency, Ep levels in the spent media of these cells reproducibly showed an exponential increase to more than 300 mU/ml at the time of saturation density. The importance of cell population density in Ep production by the renal carcinoma cell cultures was further confirmed by the observation that the cultures with higher seeding density reached confluency earlier and began an exponential increase in Ep production sooner than those cultures with lower seeding density.

Topics: Carcinoma, Renal Cell; Cell Count; Cell Division; Cells, Cultured; Erythropoietin; Humans; Kidney Neoplasms; Time Factors

Topics: Adenocarcinoma; Blood Cell Count; Carcinoma, Renal Cell; Epoetin Alfa; Erythropoietin; Geriatrics; Humans; Kidney Neoplasms; Lung Neoplasms; Mercaptopurine; Neoplasm Metastasis; Neoplasms; Neoplasms, Second Primary; Nephrectomy

Topics: Adenocarcinoma; Blood; Body Fluids; Carcinoma, Renal Cell; Epoetin Alfa; Erythrocytes; Erythropoiesis; Erythropoietin; Humans; Kidney Neoplasms; Neoplasm Metastasis; Nephrectomy; Pathology; Polycythemia; Urine

Topics: Adenocarcinoma; Bone Marrow Examination; Carcinoma, Renal Cell; Epoetin Alfa; Erythrocytes; Erythropoiesis; Erythropoietin; Hematocrit; Kidney Neoplasms; Polycythemia; Rats; Research; Tissue Extracts