epidermal-growth-factor and Carcinoma--Renal-Cell

The therapeutic landscape of renal cell carcinoma (RCC) has greatly expanded in the last decade. From being a malignancy orphan of effective therapies, kidney cancer has become today a tumor with several treatment options. Renal cell carcinoma (RCC) is a metabolic disease, being characterized by the dysregulation of metabolic pathways involved in oxygen sensing (VHL/HIF pathway alterations and the subsequent up-regulation of HIF-responsive genes such as VEGF, PDGF, EGF, and glucose transporters GLUT1 and GLUT4, which justify the RCC reliance on aerobic glycolysis), energy sensing (fumarate hydratase-deficient, succinate dehydrogenase-deficient RCC, mutations of HGF/MET pathway resulting in the metabolic Warburg shift marked by RCC increased dependence on aerobic glycolysis and the pentose phosphate shunt, augmented lipogenesis, and reduced AMPK and Krebs cycle activity) and/or nutrient sensing cascade (deregulation of AMPK-TSC1/2-mTOR and PI3K-Akt-mTOR pathways). In this complex scenario it is important to find prognostic and predictive factors that can help in decision making in the treatment of mRCC.

Topics: AMP-Activated Protein Kinases; B7-H1 Antigen; Carcinoma, Renal Cell; Epidermal Growth Factor; Fumarate Hydratase; Glucose Transporter Type 1; Glucose Transporter Type 4; Hepatocyte Growth Factor; Humans; Kidney Neoplasms; Metabolic Networks and Pathways; Molecular Targeted Therapy; Mutation; Phosphatidylinositol 3-Kinases; Platelet-Derived Growth Factor; Precision Medicine; Programmed Cell Death 1 Receptor; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-met; Receptors, Fibroblast Growth Factor; Signal Transduction; Succinate Dehydrogenase; TOR Serine-Threonine Kinases; Vascular Endothelial Growth Factor A

Major progress in treatment of renal cell carcinoma (RCC) has occurred in the past decade.. This review reports the background on the potential implication of the EGF/EGFR pathway in RCC, the different data on EGFR positivity in RCC and results from prospective Phase II and III trials on lapatinib in RCC, along with other EGF/EGFR inhibitors.. Despite important progress and the real revolution of the past decade in treatment of RCC, treatment still remains palliative most of the time. To improve treatment, it is necessary to understand whether a specific population could be selected on a molecular feature of the tumor. Until there is better knowledge of the biology, further development of drugs such as lapatinib cannot be supported.

Topics: Animals; Antineoplastic Agents; Carcinoma, Renal Cell; Clinical Trials, Phase II as Topic; Clinical Trials, Phase III as Topic; Epidermal Growth Factor; ErbB Receptors; Humans; Kidney Neoplasms; Lapatinib; Protein Kinase Inhibitors; Quinazolines

One-third of patients with renal cell carcinoma present with unresectable or metastatic disease. Immunotherapy, the current standard treatment, induces response in only 10-20% of patients. Chemotherapy with current agents is minimally effective. Other approaches including allogeneic stem cell transplant, vaccine and gene therapy and signal transduction inhibitors, offer promise in early Phase studies. This paper reviews the current treatment options and promising new agents in development.

Topics: Angiogenesis Inhibitors; Antibodies, Monoclonal; Carcinoma, Renal Cell; Cytokines; Drug Therapy; Endothelial Growth Factors; Epidermal Growth Factor; Fibroblast Growth Factors; Genetic Therapy; Hematopoietic Stem Cell Transplantation; Humans; Immunotherapy; Immunotherapy, Active; Kidney Neoplasms; Lymphokines; Transplantation, Homologous; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Metastatic renal cell carcinoma (mRCC) patients treated with anti-vascular endothelial growth factor (VEGF) therapies demonstrate promising outcomes but not all patients benefit. Factors that predict response remain to be elucidated.. Nephrectomy material from 37 patients with mRCC receiving bevacizumab +/- erlotinib was used for protein and gene expression assessment. Protein lysates were subjected to reverse-phase protein array profiling. RNA extracts were used to carry out gene expression microarray-based profiling. Normalized protein and gene expression data were correlated with overall survival (OS) and progression-free survival (PFS) using univariate Cox hazard model and linear regression. Immunoblotting was carried out to validate the results.. High protein levels of AMP-activated protein kinase and low levels of cyclin B1 (CCNB1) were associated with longer OS and PFS. Further validation revealed reduced expression and activation of phosphoinositide 3-kinase (PI3K) pathway components and cell cycle factors in patients with prolonged survival after therapy. Gene expression analysis revealed up-regulation of PI3K- and cell cycle-related pathways in patients with shorter PFS.. The OS and PFS of bevacizumab +/- erlotinib-treated patients with renal cell carcinoma were associated with changes in expression of protein and gene expression markers related to PI3K pathway and cell cycle signaling.

Topics: Angiogenesis Inhibitors; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Bevacizumab; Biomarkers, Tumor; Carcinoma, Renal Cell; Cell Cycle; Epidermal Growth Factor; Gene Expression Profiling; Humans; Kidney Neoplasms; Neoplasm Proteins; Oligonucleotide Array Sequence Analysis; Survival Analysis; Vascular Endothelial Growth Factor A

Due to a lack of knowledge of the disease process, papillary renal cell carcinoma (PRCC) has a dismal outlook. This research was aimed at uncovering the possible biomarkers and the underlying principles in PRCC using a bioinformatics method.. We searched the Gene Expression Omnibus (GEO) datasets to obtain the GSE11151 and GSE15641 gene expression profiles of PRCC. We used the R package limma to identify the differentially expressed genes (DEGs). The online tool DAVID and ClusterProfiler package in R software were used to analyze Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway dominance, respectively. The STRING database was utilized to construct the PPI network of DEGs. Using the Cytoscape technology, a protein-protein interaction (PPI) network that associated with DEGs was created, and the hub genes were identified using the Cytoscape plug-in CytoHubba. The hub genes were subjected to a Kaplan-Meier analysis to identify their correlations with survival rates.. From the selected datasets, a total of 240 common DEGs were identified in the PRCC, including 50 upregulated genes and 190 downregulated regulated genes. Renal growth, external exosome, binding of heparin, and metabolic processes were all substantially associated with DEGs. The CytoHubba plug-in-based analysis identified the 10 hub genes (. We revealed important genes and proposed biological pathways that may be implicated in the formation of PRCC. EGF might be a predictive biomarker for PRCC and therefore should be investigated as a novel treatment strategy.

Topics: Carcinoma, Renal Cell; Computational Biology; Epidermal Growth Factor; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Gene Regulatory Networks; Humans; Kidney Neoplasms

Previous studies have demonstrated that the expression of CARD10 is closely associated with the occurrence of tumors, and its role is mainly to promote tumor progression by activating the transcription factor NF‑κB. However, the signaling pathway in renal cancer remains unclear. The objective of the present study was to investigate the ability of caspase recruitment domain 10 (CARD10) to regulate the NF‑κB signaling pathway and promote the progression of renal cell carcinoma (RCC). Expression of CARD10 in ACHN, 786‑O and HK‑2 cells was evaluated via western blot analysis, as was the epidermal growth factor (EGF)‑induced activation of NF‑κB signaling pathway‑related proteins in cells. The expression of CARD10 was inhibited by CARD10 short hairpin RNA transfection. Cell cycle analysis and MTT assays were used to evaluate cell proliferation. Cell apoptosis was analyzed via flow cytometry. The invasion of renal cell lines was detected via Transwell cell migration and invasion assays in vitro. The results showed that CARD10 expression was significantly higher in RCC cells than in normal renal tubular epithelial cells. CARD10 silencing inhibited the proliferation, invasion and migration of RCC cells. EGF stimulation upregulated the activation of the NF‑κB pathway in RCC cells. Inhibition of CARD10 expression inhibited NF‑κB activation in RCC cells. Taken together, these data suggested that CARD10 promotes the progression of renal cell carcinoma by regulating the NF‑κB signaling pathway. Thus, this indicated that CARD10 may be a novel therapeutic target in RCC.

Topics: Apoptosis; Carcinoma, Renal Cell; CARD Signaling Adaptor Proteins; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Epidermal Growth Factor; Gene Expression Regulation, Neoplastic; Gene Silencing; Humans; Kidney Neoplasms; NF-kappa B; RNA, Small Interfering; Signal Transduction; Up-Regulation

Renal cell carcinoma (RCC) is the most common adult kidney cancer, and accounts for 85% of all cases of kidney cancers worldwide. Praeruptorin B (Pra-B) is a bioactive constituent of

Topics: Carcinoma, Renal Cell; Cathepsin C; Cathepsins; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Coumarins; Cysteine Endopeptidases; Epidermal Growth Factor; Gene Expression Regulation, Neoplastic; Humans; Kidney Neoplasms; MAP Kinase Signaling System; Models, Biological; Molecular Structure

Aberrant activation of the ubiquitous transcription factor STAT3 is a major driver of solid tumor progression and pathological angiogenesis. STAT3 activity is regulated by numerous posttranslational modifications (PTMs), including Tyr(705) phosphorylation, which is widely used as an indicator of canonical STAT3 function. Here, we report a noncanonical mechanism of STAT3 activation that occurs independently of Tyr(705) phosphorylation. Using quantitative liquid chromatography-tandem mass spectrometry, we have discovered and characterized a novel STAT3 phosphoform that is simultaneously phosphorylated at Thr(714) and Ser(727) by glycogen synthase kinase 3α and -β (GSK-3α/β). Both Thr(714) and Ser(727) are required for STAT3-dependent gene induction in response to simultaneous activation of epidermal growth factor receptor (EGFR) and protease-activated receptor 1 (PAR-1) in endothelial cells. In this combinatorial signaling context, preventing formation of doubly phosphorylated STAT3 by depleting GSK-3α/β is sufficient to disrupt signal integration and inhibit STAT3-dependent gene expression. Levels of doubly phosphorylated STAT3 but not of Tyr(705)-phosphorylated STAT3 are remarkably elevated in clear-cell renal-cell carcinoma relative to adjacent normal tissue, suggesting that the GSK-3α/β-STAT3 pathway is active in the disease. Collectively, our results describe a functionally distinct, noncanonical STAT3 phosphoform that positively regulates target gene expression in a combinatorial signaling context and identify GSK-3α/β-STAT3 signaling as a potential therapeutic target in renal-cell carcinoma.

Topics: Carcinoma, Renal Cell; Cells, Cultured; Early Growth Response Protein 1; Epidermal Growth Factor; ErbB Receptors; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Human Umbilical Vein Endothelial Cells; Humans; Kidney Neoplasms; Phosphorylation; Promoter Regions, Genetic; Protein Binding; Protein Processing, Post-Translational; Receptor, PAR-1; Signal Transduction; STAT3 Transcription Factor; Transcriptional Activation

The epidermal growth factor (EGF) is responsible for the activation of intracellular signal transducers that act on cell-cycle progression, cell motility, angiogenesis and inhibition of apoptosis. However, cells can block these effects activating opposite signaling pathways, such as the transforming growth factor beta 1 (TGFβ1) pathway. Thus changes in expression levels of EGF and TGFB1 in renal cells might modulate the renal cell carcinoma (RCC) development, in consequence of changes in regulatory elements of signaling networks such as the microRNAs (miRNAs). Our purpose was to investigate the synergic role of EGF+61G>A and TGFB1+869T>C polymorphisms in RCC development. Genetic polymorphisms were studied by allelic discrimination using real-time PCR in 133 RCC patients vs. 443 healthy individuals. The circulating EGF/EGFR-MAPK-related miR-7, miR-221 and miR-222 expression was analyzed by a quantitative real-time PCR in plasma from 22 RCC patients vs. 27 healthy individuals. The intermediate/high genetic proliferation profile patients carriers present a significantly reduced time-to-progression and a higher risk of an early relapse compared with the low genetic proliferation profile carriers (HR = 8.8, P = 0.038) with impact in a lower overall survival (Log rank test, P = 0.047). The RCC patients presented higher circulating expression levels of miR-7 than healthy individuals (6.1-fold increase, P<0.001). Moreover, the intermediate/high genetic proliferation profile carriers present an increase in expression levels of miR-7, miR-221 and miR-222 during the RCC development and this increase is not observed in low genetic proliferation profile (P<0.001, P = 0.004, P<0.001, respectively). The stimulus to angiogenesis, cell-cycle progression and tumoral cells invasion, through activation of EGFR/MAPK signaling pathway in intermediate/high proliferation profile carriers is associated with an early disease progression, resulting in a poor overall survival. We also demonstrated that the intermediate/high proliferation profile is an unfavorable prognostic factor of RCC and miR-7, miR-221 and miR-222 expressions may be useful phenotype biomarkers of EGFR/MAPK activation.

Topics: Adult; Carcinoma, Renal Cell; Epidermal Growth Factor; Humans; Kidney Neoplasms; MicroRNAs; Middle Aged; Polymorphism, Single Nucleotide; Signal Transduction; Survival Analysis; Transforming Growth Factor beta1

To identify a phenotype that could be informative and prognostic in patients with renal cell carcinoma (RCC) peripheral blood was evaluated for TH1, TH2, regulatory T cells (Tregs), natural killer (NK) and NKT cells and for cytokines/chemokines.. Peripheral blood from 77 patients with RCC and 40 healthy controls was evaluated by flow cytometry using monoclonal antibodies against CD4, CD25, FoxP3, CD45RA, CD45RO, CD152, CD184, CD279, CD3, CD16, CD56, CD161, CD158a, CD4, CD26, CD30, CD183 and CD184. A concomitant evaluation of 38 molecules was conducted in patients' serum using a multiplex biometric ELISA-based immunoassay.. The number of NK cells CD3⁻/CD16⁺, CD3⁻/CD16⁺/CD161⁺ (NK) and CD3⁻/CD16⁺/CD161⁺/CD158a⁺ (NK- Kir 2+) was greater in the patients with RCC (P < 0.05); and the number of Treg cells CD4⁺/CD25(high+)/FOXP3⁺ and the subset CD4⁺/CD25(high+)/FOXP3⁺/CD45RA⁺ (naïve) and CD45R0⁺(memory) cells, were greater in the patients with RCC (P < 0.001). An increase in the following was observed in the serum of patients with RCC compared with healthy controls: interleukin (IL)-4, IL-6, IL-8, IL-10, G-CSF, CXCL10, CXCL11, hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF). According to Ingenuity Pathway Analysis (IPA), CXCL10, IL-6, IL-8, epidermal growth factor (EGF), HGF and VEGF were associated with a network that controls cellular movement, tissue development and cellular growth. Kaplan-Meier analysis for disease-free survival showed that high numbers of CD4⁺/CD25(high+)/FOXP3⁺/CD45RA⁺ (Treg naïve) and low numbers of CD3⁻/CD16⁺/CD161⁺/CD158a⁺ (NK-Kir+) cells predict short disease-free survival in patients with RCC.. Concomitant evaluation of Treg (CD4⁺/CD25(high+)/FOXP3⁺ and CD4⁺/CD25(high+)/FOXP3⁺/CD45RA⁺) and of six soluble factors (IL-6, IL-8 ,VEGF, CXCL10, CXCL11, EGF, HGF) might be a surrogate marker of host immunity in patients with RCC.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Carcinoma, Renal Cell; CD4-Positive T-Lymphocytes; Chemokine CXCL10; Chemokine CXCL11; Disease Progression; Disease-Free Survival; Epidermal Growth Factor; Female; Flow Cytometry; Hepatocyte Growth Factor; Humans; Immunophenotyping; Interleukin-6; Interleukin-8; Kaplan-Meier Estimate; Kidney Neoplasms; Killer Cells, Natural; Male; Middle Aged; T-Lymphocytes, Regulatory; Vascular Endothelial Growth Factor A

The epidermal growth factor (EGF) gene encodes a growth factor that binds to the EGF receptor (EGFR), which is involved in activating pathways that promote cellular proliferation, survival, migration and differentiation, and lack of control is characteristic of malignant development. Previous studies showed that serum EGF levels may influence the risk of cancer. In this study, we genotyped the EGF G61A polymorphism (rs4444903) and measured serum EGF levels using an enzyme immunoassay in a hospital-based case-control study of 345 patients with diagnosed renal cell carcinoma (RCC) and 346 cancer-free controls in a Chinese population. Compared with the EGF 61GG genotype, the AA genotype had a significantly increased RCC risk (odds ratio=1.80, 95% confidence interval=1.04-3.12). Besides, the mean serum EGF levels in RCC patients (858.94+/-391.54 pg ml(-1)) were significantly lower than those in controls (1281.52+/-568.42 pg ml(-1), P<0.001). In addition, individuals carrying AA genotype had lower serum EGF levels than GA or GG carriers. These results suggested that the EGF G61A polymorphism is involved in the etiology of RCC and thus may be a marker for genetic susceptibility to RCC in Chinese populations. Larger studies are warranted to validate our findings.

Topics: Adult; Aged; Asian People; Body Mass Index; Carcinoma, Renal Cell; Case-Control Studies; China; Epidermal Growth Factor; Female; Gene Frequency; Genetic Predisposition to Disease; Genotype; Humans; Kidney Neoplasms; Logistic Models; Male; Middle Aged; Polymorphism, Single Nucleotide; Risk Factors; Smoking

Various types of malignant tumor have been found to contain a subpopulation of cancer stem cells (CSCs). In this study, we sought to enrich CSCs from the renal cell carcinoma (RCC) cell line SK-RC-42 using the sphere culture system and characterize their immunophenotype. We demonstrated that a subpopulation of SK-RC-42 cells were capable of growing as tumor spheres in serum-free medium supplemented with EGF and bFGF. The sphere-forming cells (SFCs) had many properties similar to CSCs: ability of self-renewing in vitro and in vivo, higher mRNA expression levels of several 'stemness' genes, stronger tumorigenicity and resistance to chemotherapeutic agents and irradiation compared with the monolayer adherent cells (MACs). The SFCs expressed high levels of MHC class I but low levels of MHC class II, CD80 and CD86. In contrast with MACs, the SFCs had lower expression levels of FasL and Fas, Her2 and hTERT and activating natural killer receptors. Finally, SK-RC-42 SFCs and MACs both expressed significant and comparable levels of the transcription factor forkhead box protein 3 (FoxP3) and membrane complement regulatory proteins (mCRPs). Taken together, these findings indicate that CSCs can be enriched from RCC by culturing the tumor cells as spheres. The immunophenotype of the SFCs demonstrated in this study suggests that CSCs might play an important role in the evasion of tumor growth from immune surveillance.

Topics: Animals; Antigens, CD; Antimetabolites, Antineoplastic; Carcinoma, Renal Cell; Cell Adhesion; Cell Culture Techniques; Cell Line, Tumor; Cell Proliferation; Cell Survival; Endoglin; Epidermal Growth Factor; Fibroblast Growth Factor 2; Flow Cytometry; Fluorouracil; Gene Expression Regulation, Neoplastic; Humans; Immunophenotyping; Kidney Neoplasms; Mice; Mice, Nude; Neoplasms, Experimental; Neoplastic Stem Cells; Octamer Transcription Factor-3; Receptors, Cell Surface; Reverse Transcriptase Polymerase Chain Reaction; Spheroids, Cellular; Transplantation, Heterologous

Topics: Antineoplastic Agents; Carcinoma, Renal Cell; Epidermal Growth Factor; Humans; Kidney Neoplasms

Renal cell carcinoma (RCC) frequently produces metastases to the musculoskeletal system that are a major source of morbidity in the form of pain, immobilization, fractures, neurological compromise, and a decreased ability to perform activities of daily living. Patients with metastatic RCC therefore have a dismal prognosis because there is no effective adjuvant treatment for this disease. Because the epidermal growth factor receptor (EGF-R) signaling cascade is important in the growth and metastasis of RCC, its blockade has been hypothesized to inhibit tumor growth and hence prevent resultant bone destruction. We determined whether blockade of EGF-R by the tyrosine kinase inhibitor PKI 166 inhibited the growth of RCC in bone. We use a novel cell line, RBM1-IT4, established from a human RCC bone metastasis. Protein and mRNA expression of the ligands and receptors was assessed by Western and Northern blots. The stimulation of RBM1-IT4 cells with epidermal growth factor or transforming growth factor alpha resulted in increased cellular proliferation and tyrosine kinase autophosphorylation. PKI 166 prevented these effects. First, RBM1-IT4 cells were implanted into the tibia of nude mice, where they established lytic, progressively growing lesions, after which the mice were treated with PKI 166 alone or in combination with paclitaxel (Taxol). Immunohistochemical analysis revealed that tumor cells and tumor-associated endothelial cells in control mice expressed activated EGF-R. Treatment of mice with PKI 166 alone or in combination with Taxol produced a significant decrease in the incidence and size of bone lesions as compared with the results in control or Taxol-treated mice (P < 0.001). Treatment with PKI 166 also decreased the expression of phosphorylated EGF-R by tumor cells and tumor-associated endothelial cells, and this was even more pronounced with PKI 166 plus Taxol treatment. The PKI 166 plus Taxol combination produced apoptosis of tumor cells and tumor-associated endothelial cells. Tumor cell proliferation, shown by proliferating cell nuclear antigen positivity, was decreased in all treatment groups. In addition, the integrity of the bone was maintained in mice treated with PKI 166 or PKI 166 plus Taxol, whereas massive bone destruction was seen in control and Taxol-treated mice. These results suggest that blockade of EGF-R signaling inhibits growth of RCC in the bone by its effect on tumor cells and tumor-associated endothelial cells.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Bone Neoplasms; Carcinoma, Renal Cell; Cell Division; Epidermal Growth Factor; ErbB Receptors; Humans; Kidney Neoplasms; Male; Mice; Mice, Nude; Paclitaxel; Phosphorylation; Pyrimidines; Pyrroles; Recombinant Proteins; Signal Transduction; Transforming Growth Factor alpha; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Previous experiments have shown that both in vivo and in vitro pre-treatment with various hormones increases the renal transport capacity for weak organic acids, such as PAH, in rats. The aim of the present study was to test whether or not accumulation of the anticancer drugs methotrexate (MTX), cisplatin (CP), raltitrexed (Tomudex) and topotecan (Hycamtin) can be increased in intact, healthy rat and human renal cortical slices and in human renal cell carcinoma (RCC). Intact, healthy human tissue was obtained from tumour bearing kidneys of patients suffering from RCC. Experiments were intended as a new approach to overcome so-called multidrug resistance. Kidney tissue slices were incubated for 24 h in William's medium E containing various concentrations of dexamethasone, T(3), or EGF. Thereafter slices were placed in anticancer drug containing Cross-Taggart medium and the drug uptake into kidney tissue was measured for 2 h. In intact rat and human renal tissue slices, the uptake of p-aminohippurate (PAH = reference substance) increased significantly after incubation in dexamethasone containing medium (134% and 156%, respectively). There were no stimulating effects of either T(3) or EGF on PAH accumulation. On the other hand, only the accumulation of MTX, but not of CP, raltitrexed or topotecan, was significantly enhanced after hormone pre-treatment both in intact renal tissue and in RCC. A stimulation of renal PAH accumulation can be performed ex vivo, as reported previously, both in intact rat and human renal cortical slices and in RCC. Discrepancies between the effects of dexamethasone and T(3) or EGF indicate different modes of action of these substances at the cellular level. Unfortunately, with the exception of MTX, the uptake of anticancer drugs can not be stimulated effectively ex vivo in human RCC tissue by the substances used. Evidently the transport of these anticancer drugs out of the kidney cells is more effective than their uptake.

Topics: Animals; Antimetabolites, Antineoplastic; Antineoplastic Agents; Biological Transport; Carcinoma, Renal Cell; Cisplatin; Dexamethasone; Epidermal Growth Factor; Female; Glucocorticoids; Humans; Kidney; Kidney Neoplasms; Methotrexate; Quinazolines; Rats; Rats, Wistar; Thiophenes; Topotecan; Triiodothyronine

We compared cytokine levels in fluid from renal cysts with and without renal cell carcinoma.. Fluid was aspirated from 18 renal cysts without (benign) and 21 with renal cell carcinoma (malignant). Serum from patients with renal cell carcinoma and healthy controls was collected and cytokines were measured by enzyme-linked immunosorbent assay.. Interleukin-6 (IL-6) and basic fibroblast growth factor concentrations were higher in malignant than benign cysts or serum (p <0.006). Epidermal growth factor levels were significantly higher in malignant cysts and serum than in benign cysts (p <0.01). IL-6 levels in malignant cysts positively correlated with the erythrocyte sedimentation rate (R=0.80) and C-reactive protein (R=0.86), and they were higher in grade 3 than in grade 2 tumors. Basic fibroblast growth factor levels were significantly higher in malignant cysts associated with hypervascular than hypovascular tumors (p=0.029).. Cytokine levels in aspirated fluid may help to identify malignant renal cysts and indicate the characteristics of coexisting tumors.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Carcinoma, Renal Cell; Cytokines; Epidermal Growth Factor; Female; Fibroblast Growth Factor 2; Humans; Interleukin-6; Kidney Diseases, Cystic; Kidney Neoplasms; Male; Middle Aged

Human renal cell carcinoma (RCC) tissue and a cell line derived therefrom, SMKT-R3, showed markedly increased glycolipid sulfotransferase [cerebroside sulfotransferase (CST); EC 2.8.2.11] activity and accumulated sulfoglycolipids. Recently, we cloned a human CST cDNA from a SMKT-R3 cDNA library (K. Honke et al., J. Biol. Chem., 272: 4864-4868, 1997). In this study, we investigated the expression of the CST gene in seven human RCC lines (SMKT-R1, SMKT-R2, SMKT-R3, SMKT-R4, TOS-1, TOS-2, and ACHN) and their normal counterpart, human renal proximal tubular cells. On Northern blot analysis, a marked increase of CST mRNA was observed in every RCC line, except for ACHN, as compared with normal cells. ACHN cells showed a slightly increased level of CST mRNA. CST activity was correlated with the amount of mRNA. Sulfoglycolipid analysis revealed that expression of lactosylceramide sulfate was correlated with the CST level. Furthermore, we examined the effects of epidermal growth factor (EGF), tetradecanoylphorbol-13-acetate, and genistein, which are known to regulate CST activity in SMKT-R3 cells, on CST-gene expression in various RCC cells. On treatment with EGF, CST mRNA time-dependently increased in accord with its activity in SMKT-R3 cells. Yet, augmentation by EGF was only observed in SMKT-R3. In contrast, a reduction of CST mRNA and activity by tetradecanoylphorbol-13-acetate and genistein was observed in all of the lines examined. Taken together, these findings indicate that in human RCC cells, the CST gene is generally overexpressed via a signaling pathway involving protein kinase-C and tyrosine kinases.

Topics: Carcinoma, Renal Cell; Epidermal Growth Factor; Gene Expression Regulation, Enzymologic; Humans; Kidney Neoplasms; Kidney Tubules, Proximal; Protein-Tyrosine Kinases; RNA, Messenger; Sulfotransferases; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

We demonstrate the constitutive expression of interleukin 6 (IL-6), IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), epidermal growth factor (EGF), transforming growth factor alpha (TGF-alpha) and epidermal growth factor receptor (EGFR) in normal kidney cells, and in the majority of renal oncocytomas, papillary and non-papillary renal cell carcinomas (RCCs) by reverse transcriptase polymerase chain reaction (RT-PCR) technique. No expression of IL-6 and TGF-alpha and variable expression of GM-CSF, IL-8, EGF and EGFR was seen in chromophobe RCCs. The lack of expression of IL-6 and TGF-alpha might be correlated with the growth pattern, poor vascularity and low malignancy of chromophobe RCCs.

Topics: Carcinoma, Papillary; Carcinoma, Renal Cell; Epidermal Growth Factor; ErbB Receptors; Gene Expression; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-6; Kidney Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor alpha

The aim of the present study was to analyze the contribution of different stimulatory and inhibitory growth factors to the deregulated proliferation of human RCCs.. The expression of different growth factors and their corresponding receptors were analyzed by Northern blot, FACS, ELISA and immunocytochemistry in 13 permanent human RCC cell lines of the clear cell type. Moreover, the functional intactness of growth factor-related signal transduction pathways was investigated.. All RCC cell lines expressed EGF-receptor mRNA and protein and 10 cell lines secreted TGF-alpha. Exogeneously added TGF-alpha resulted in a significant (p < 0.05) stimulation of growth in 6 RCC cell lines and a significant (p < 0.05) inhibition of proliferation in 3 cell lines. PDGF B and the corresponding type beta receptor were expressed in a single cell line. mRNA expression of PDGF A and PDGF-alpha-receptor as well as IGF-1 and its receptor could not be detected in any cell line. Eleven RCC cell lines expressed TGF-beta 1 mRNA and in all cell lines TGF-beta 1 secretion into the supernatant could be demonstrated. Whereas all cell lines exhibited TGF-beta type II-receptor mRNA, type I-receptor mRNA could be detected only in 3 cell lines. TGF-beta type III-receptor was observed in 1 cell line. Exogeneously added TGF-beta1 resulted in a significant (p < 0.05) inhibition of proliferation in 7 RCC cell lines.. Clear cell RCCs exhibit a complex and heterogeneous expression pattern for various growth factors and their receptors. Growth factor secretion and intact signal transduction pathways in most clear cell RCCs facilitate an intricate modulation of RCC growth by autocrine and paracrine interactions between tumor cells and host tissue.

Topics: Carcinoma, Renal Cell; Cell Division; Epidermal Growth Factor; ErbB Receptors; Humans; Kidney Neoplasms; Platelet-Derived Growth Factor; Receptor, IGF Type 1; Receptors, Platelet-Derived Growth Factor; RNA; Transforming Growth Factor alpha; Transforming Growth Factor beta; Tumor Cells, Cultured

Glycolipid sulfotransferase activity in a human renal cell carcinoma cell line, SMKT-R3, is enhanced by epidermal growth factor (EGF); tyrosine kinase inhibitors suppress this enhancement. To investigate the involvement of Ras in the signal transduction pathway from the EGF receptor to the expression of glycolipid sulfotransferase, we introduced v-H-ras into SMKT-R3 cells. In a quiescent state, the percent GTP bound to Ras in v-H-ras-expressing cells increased about 2.5-fold compared with control cells, suggesting that v-Ras introduced into the renal cancer cells is in an active form without EGF stimulation. Glycolipid sulfotransferase activity in v-H-ras-expressing cells was higher than in control cells. The sulfotransferase activity was affected neither by EGF nor by genistein, a tyrosine kinase inhibitor, in v-H-ras-expressing cells, whereas it was enhanced by EGF and reduced by genistein in control cells. Our observations suggest that Ras mediates the regulation pathway of glycolipid sulfotransferase activity in SMKT-R3 cells.

Topics: Carcinoma, Renal Cell; Enzyme Induction; Enzyme Inhibitors; Epidermal Growth Factor; Genes, ras; Genistein; Humans; Isoflavones; Kidney Neoplasms; Neoplasm Proteins; Oncogene Protein p21(ras); Protein-Tyrosine Kinases; Signal Transduction; Sulfotransferases; Transfection

Expression and prognostic impact of some exponents of the epidermal growth factor (EGF) family in renal cell carcinoma (RCC) were examined.. EGF, transforming growth factor-alpha (TGF-alpha), EGF receptor (EGF-R), and c-erb B-2 were determined immunohistochemically in formalin-fixed paraffin-embedded tumor samples of 30 patients with locally confined RCCs. The prognostic significance of these growth factors and their receptors as well as of tumor stage and malignancy grade was examined with respect to survival and tumor recurrence by following up the fate of the patients after nephrectomy (mean follow-up time 5.2 years).. The members of the EGF family and their receptors studied were expressed to a variable degree in all RCCs investigated. However, using log-rank tests in Kaplan-Meier plots only tumor stage (p < 0.0007) and malignancy grade (p < 0.007) but none of the growth factors or receptors studied (p > 0.05, respectively) exhibited prognostic significance with respect to both survival and disease-free period. On the contrary, there was a significant correlation between EGF and TGF-alpha (p < 0.001), EGF and EGF-R (p = 0.028), EGF-R and c-erb B-2 (p = 0.0009), and-inversely related-between TGF-alpha and tumor stage (p = 0.047) and between EGF-R and malignancy grade (p = 0.03). The coexpression of the factors studied also showed no prognostic relevance.. The expression of these members of the EGF family seems not to bear evaluable prognostic information for clinical use in the case of RCC.

Topics: Adult; Aged; Carcinoma, Renal Cell; Epidermal Growth Factor; ErbB Receptors; Female; Follow-Up Studies; Gene Expression Regulation, Neoplastic; Genes, erbB-2; Humans; Immunohistochemistry; Kidney Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Nephrectomy; Prognosis; Receptor, ErbB-2; Retrospective Studies; Transforming Growth Factor alpha

We studied the association between epidermal growth factor (EGF) binding capacity and histopathologic features or prognosis in human renal cell carcinoma (RCC) by Scatchard analysis in 67 patients. EGF binding capacity was significantly greater in metastatic than in nonmetastatic tumors, and in nuclear grade 3 than nuclear grade 1 tumors. Multivariate analysis revealed that tumor stage, nuclear grade, EGF binding capacity, and tumor size significantly correlated with overall survival. These results suggest that EGF binding may be an important determinant of prognosis in patients with RCC.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Renal Cell; Epidermal Growth Factor; Female; Humans; Kidney Neoplasms; Male; Middle Aged; Multivariate Analysis; Prognosis; Prospective Studies; Survival Analysis

Renal cell carcinoma (RCC) is thought to arise from the renal tubular cells (RTC). Assuming that proliferating RTC imply a premalignant change of RTC into RCC, messenger RNA expressions of growth factors in cultured RTC were compared to both cultured and frozen noncultured RCC.. The expression of transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF), EGF receptor (EGFR) and interleukin-6 (IL-6) were studied in surgically obtained RCC (n = 17), cultured RCC (n = 10), and autologous cultured RTC (n = 15). Quantitation of the PCR product was performed using a computer image analyzer which evaluated the intensity of each cytokine relative to beta-actin.. TGF-alpha, EGFR and IL-6 were detected in most of the cultured RTC, and both cultured and noncultured RCC were also expressed at high levels. In contrast to a high positivity of TGF-alpha, EGF was not strongly positive in all specimens.. Our results show that there is a predominant autocrine production of TGF-alpha in RCC and RTC, suggesting that TGF-alpha plays a distinct role in the proliferation of these cells. These studies also indicate that the mechanisms of proliferation and cytokine production of RCC and RTC are similar.

Topics: Carcinoma, Renal Cell; Cell Division; Epidermal Growth Factor; Gene Expression; Humans; Immunohistochemistry; Interleukin-6; Kidney Neoplasms; Kidney Tubules; Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Transforming Growth Factor alpha; Tumor Cells, Cultured

In the previous studies, epidermal growth factor (EGF) was found to elevate glycolipid sulfotransferase activity in a human renal cell carcinoma cell line, SMKT-R3. To elucidate whether Ras is involved in the signal transduction pathway from EGF to the expression of the sulfotransferase, effects of EGF and a tyrosine kinase inhibitor on the sulfotransferase activity were investigated in renal cancer cells stably expressing activated Ras. SMKT-R3 cells were transfected with a plasmid carrying v-H-ras (pv-H-ras). As a control, SMKT-R3 cells transfected with a mutant v-H-ras, in which glycine at position 15 was replaced by valine (pG15V), was used. The expression of v-H-ras in transfected cells was examined by RT-PCR analysis. Two clones transfected with pv-H-ras, named A1 and A6, and two clones transfected with pG15 V, termed B1 and B4, were found to express v-H-ras and G15V genes, respectively, and employed in the following experiments. Though EGF elevated the activity of glycolipid sulfotransferase in B1, B4 and SMKT-R3 cells, it did not change the activity levels in A1 and A6 cells. This result suggested that since the signal from Ras was saturated in the cells expressing activated Ras, the cells did not respond to the stimulation by EGF. To know the association between tyrosine kinase of EGF receptor and Ras, the effect of Genistein, a specific inhibitor of tyrosine kinase, on the sulfotransferase activity was examined in these cell lines. Genistein reduced the activity of glycolipid sulfotransferase in B1, B4, and SMKT-R3 cells, whereas it hardly affected the sulfotransferase activity in A1 and A6 cells. This result indicated that the signal from tyrosine kinase was dissociated from the regulation of the sulfotransferase activity in the cells expressing activated Ras. These observations suggest that Ras is involved in the signal transduction from EGF to the expression of glycolipid sulfotransferase activity in SMKT-R3 cells and acts in the downstream of tyrosine kinase of EGF receptor.

Topics: Amino Acid Sequence; Base Sequence; Carcinoma, Renal Cell; Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation, Enzymologic; Genes, ras; Genistein; Humans; Isoflavones; Kidney Neoplasms; Molecular Sequence Data; Protein-Tyrosine Kinases; Signal Transduction; Sulfotransferases; Transfection; Tumor Cells, Cultured

In order to quantitate epidermal growth factor receptors (EGFR) in human renal cell carcinoma (RCC) tissues, the binding of isotope-labeled epidermal growth factor (125I-EGF) to pooled membrane samples from human RCC was studied. Specific EGF binding to membranes at 4 degrees C reached a plateau after 2 h of incubation and remained constant up to 8 h; specific binding at 25 degrees C reached a plateau after 30 min of incubation, then decreased gradually. 125I-EGF binding to the membrane was displaced by both EGF and transforming growth factor-alpha, but not by insulin and basic fibroblast growth factor. Scatchard analysis of the specific EGF binding generated a straight line, indicating a single class of binding sites for EGF, with a dissociation constant (Kd) of 21.1 x 10(-10) M and a maximum number of binding sites of 57.2 fmol/mg membrane protein. When the protein concentration in the incubation medium was adjusted from 0.71 mg/ml to 2.84 mg/ml, Scatchard analysis revealed identical Kd values, and the maximum number of binding sites was proportional to the protein concentration. These results demonstrate the presence of EGFR on RCC membranes and indicate that these receptors can be studied quantitatively.

Topics: Binding Sites; Biological Assay; Carcinoma, Renal Cell; Cell Membrane; Data Interpretation, Statistical; Epidermal Growth Factor; ErbB Receptors; Humans; Kidney Neoplasms; Protein Binding; Proteins; Sensitivity and Specificity

Epidermal growth factor (EGF), a potent mitogen and tumor promoter, is excreted in urine permitting it to incubate with urothelial cells. We have previously shown that the distribution of receptors for EGF (EGF-Rs) on malignant urothelium changes to favor contact between EGF-Rs and intraluminal EGF. OBJECTIVES. To determine if concentrations of EGF in voided urine are different in patients with transitional cell carcinoma (TCC) of the bladder than in those without this condition. METHODS. EGF was measured by radioimmunoassay in the urine of 54 patients with newly diagnosed TCC (16 with grade 1, Stage Ta lesions, and 38 with grade 3, Stage T > or = 2 lesions) and 66 pathologic and normal controls without TCC. Subjects were matched for age and good renal function. RESULTS. EGF concentrations were significantly reduced in patients with TCC compared with controls either when uncorrected (p < 0.0001) or corrected (p < 0.000000002) for urinary creatinine excretion. CONCLUSIONS. The reduced concentration of EGF in voided urine supports previous evidence that the urinary EGF/urothelial EGF-R interaction is important for TCC development and growth, and has the potential to serve as a marker of tumor persistence, recurrence, and therapeutic response.

Topics: Age Factors; Aged; Biomarkers, Tumor; Carcinoma, Renal Cell; Carcinoma, Transitional Cell; Case-Control Studies; Creatinine; Epidermal Growth Factor; Female; Humans; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Regression Analysis; Sex Factors; Urinary Bladder Neoplasms

The expression of epidermal growth factor receptor (EGFR) mRNA has been demonstrated in human renal cell carcinomas (RCC), but few reports quantitate EGFR in RCC and correlate EGFR content with clinicopathologic findings.. Using 125I-EGF as a ligand, the maximum binding of EGF in membrane preparations from RCC (EGFR content) was studied by Scatchard analysis.. A single class of binding sites for EGF was observed in 74% of RCC and 50% of normal renal tissues. The EGFR content was increased significantly in RCC compared with normal tissues. In all cases in which EGFR was undetectable, there was no evidence of distant metastasis, venous invasion, or regional lymph node involvement, and these patients had a better clinical outcome than patients with detectable EGFR. The EGFR content was significantly lower in nuclear Grade 1 tumors than in tumors of higher nuclear grades. No significant difference between EGFR content and other clinicopathologic findings was detected.. The determination of EGFR content in RCC may become an important prognostic factor for the biologic behavior of RCC.

Topics: Adult; Age Factors; Aged; Aged, 80 and over; Carcinoma, Renal Cell; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Iodine Radioisotopes; Kidney; Kidney Neoplasms; Male; Middle Aged; Neoplasm Staging; Prognosis

We investigated the influence of epidermal growth factor (EGF) and transforming growth factor (TGF)-beta 1 on the in vitro invasion and type IV collagenolytic activity of two new cell lines of renal cell carcinoma (SRCC-1P and SRCC-1M). When cells were treated with EGF or with TGF-beta 1, EGF increased the number of cells penetrating through the reconstituted basement membrane in SRCC-1P. In contrast, TGF-beta 1 suppressed the number of cells penetrating through the membrane in SRCC-1M. In accordance with the invasiveness, EGF enhanced the activity of type IV collagenolysis in SRCC-1P, and TGF-beta 1 suppressed it in SRCC-1M. The growth factors did not affect DNA synthesis of the cells as evaluated by 3H-thymidine incorporation. These results suggest that EGF and TGF-beta 1 can influence the in vitro invasive process of renal cell carcinoma cells through their actions on proteolysis such as type IV collagenolysis.

Topics: Carcinoma, Renal Cell; Collagen; DNA, Neoplasm; Epidermal Growth Factor; Humans; Kidney Neoplasms; Neoplasm Invasiveness; Transforming Growth Factor beta; Tumor Cells, Cultured

O-Phospho-L-tyrosine (P-Tyr), a substrate for a wide range of protein tyrosine phosphatases, inhibited growth of human renal and breast carcinoma cells. Growth was blocked in the S phase of the cell cycle. A decrease in the amount of cyclin proteins A and B was also observed. P-Tyr incubation led to activation of cellular protein tyrosine phosphatases resulting in the inhibition of tyrosine phosphorylation of epidermal growth factor receptor as well as of p34cdc2. P-Tyr synergistically sensitized the renal carcinoma ACHN cells to killing by the chemotherapeutic agents doxorubicin and etoposide. These growth inhibitory properties of P-Tyr in vitro suggest its possible use as an anticancer agent.

Topics: Breast Neoplasms; Carcinoma, Renal Cell; Cell Death; Cell Division; Doxorubicin; Enzyme Activation; Epidermal Growth Factor; ErbB Receptors; Etoposide; Humans; Kidney Neoplasms; Neoplasm Proteins; Phosphorylation; Phosphotyrosine; Protein Tyrosine Phosphatases; S Phase; Sensitivity and Specificity; Tumor Cells, Cultured; Tyrosine

Accumulation of sulfoglycolipids associated with markedly elevated levels of glycolipid-sulfotransferase activity was previously demonstrated in human renal-cell-carcinoma cells. To elucidate the regulatory mechanisms of sulfoglycolipid metabolism in renal-cell carcinoma, effects of various growth factors on the sulfotransferase-activity levels were investigated using a human renal-cell-carcinoma cell line, SMKT-R3. Exogenous epidermal growth factor (EGF) significantly increased the activity levels of the sulfotransferases in a dose-dependent manner, but did not change that of arylsulfatase A, which hydrolyzes sulfoglycolipids. Furthermore, metabolic labeling with 35S-sulfate revealed that the addition of EGF to the culture medium of the cells resulted in an increment of sulfoglycolipid synthesis. The expression of the EGF receptor on SMKT-R3 cells was demonstrated by affinity cross-linking with 125I-EGF. These observations suggest that EGF can regulate sulfotransferase-activity levels in renal-cell-carcinoma cells, and function as one of the regulatory factors of sulfoglycolipid synthesis in these carcinoma cells.

Topics: Carcinoma, Renal Cell; Cerebroside-Sulfatase; Epidermal Growth Factor; ErbB Receptors; Glycolipids; Humans; Kidney Neoplasms; Sulfotransferases; Sulfurtransferases; Tumor Cells, Cultured

A cell line (SMKT-R3) established from renal cell carcinoma was characterized by sulfolipids and glycolipid sulfotransferases. It was found by analyzing glycolipids extracted from SMKT-R3 cells that sulfolipids constituted a large part of acidic glycolipids. When SMKT-R3 cells were metabolically labeled with sodium [35S]sulfate, the incorporation of the radioactivity was detected in accordance with SM4, SM3 and SM2, which were major sulpholipids found in the cells, by autoradiography of thin layer chromatogram of the acidic glycolipid extracts from the cells. Markedly high activity level of glycolipid sulfotransferases toward GalCer and LacCer as substrates was observed in SMKT-R3 cells. Effects of 12-0-tetradecanoylphorbol-13-acetate (TPA) and a protein kinase C inhibitor on glycolipid sulfotransferases were investigated in SMKT-R3 cells. The treatment with TPA caused a dose- and a time-dependent reduction of the enzymes activities. Similarly, H-7 and staurosporine, which are inhibitors of protein kinase C, reduced the glycolipid sulfotransferase activities. These results indicate that the glycolipid sulfotransferase activities are mediated by protein kinase C in SMKT-R3 cells. On the other hand, the treatment of SMKT-R3 cells with epidermal growth factor (EGF) was associated with the increase of the glycolipid sulfotransferase activities in a dose-dependent manner. However, this effect of EGF was counteracted by the pretreatment with TPA or H-7. These findings suggest that EGF induces the glycolipid sulfotransferase activities through protein kinase C.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Alkaloids; Carcinoma, Renal Cell; Epidermal Growth Factor; Humans; Isoquinolines; Kidney Neoplasms; Lipid Metabolism; Lipids; Piperazines; Protein Kinase C; Staurosporine; Sulfotransferases; Sulfurtransferases; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

A specific heterogeneous enzyme-linked immunosorbent assay (ELISA) has been established in order to determine levels of low-molecular-weight epidermal growth factor (EGF) in the urine of patients with renal cell carcinoma who had undergone unilateral radical nephrectomy. Urine specimens, i.e., 20 pre- and postsurgical specimens from a group of patients and 22 from a control group, were assayed after the urine had been freed from high-molecular-weight proteins (greater than 30 kDa) and salts. EGF levels were expressed as urinary EGF/creatinine ratios, and a highly significant decrease (alpha = 0.0005 by Student's t-test) of urinary EGF was found in the patient group prior to surgery. The cancer patients also showed an additional loss of urinary EGF after unilateral nephrectomy (alpha = 0.0005 by Student's t-test). These data correlate with our previous findings that pro-EGF gene expression is decreased in human renal carcinoma and support the concept that low-molecular-weight urinary EGF is derived from high-molecular-weight kidney pro-EGF.

Topics: Biomarkers, Tumor; Carcinoma, Renal Cell; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Female; Gene Expression; Humans; Kidney Neoplasms; Male; Middle Aged

Findings of increased numbers of epidermal growth factor receptors (EGF-R) and increased expression of transforming growth factor alpha (TGF-alpha) in surgical specimens of human renal cell carcinoma have led to the proposal that growth of these tumors may be regulated by TGF-alpha in an autocrine manner. In the studies presented here, we have examined this hypothesis using two human renal carcinoma cell lines, SKRC-4 and SKRC-29. We demonstrated that both SKRC-4 and SKRC-29 cells were growth stimulated by greater than 35% when cultured in the presence of TGF-alpha or EGF and were inhibited by 29% to 46% if cultured in the presence of anti-EGF-R monoclonal antibody 225. Treatment of cells with TGF-alpha enhanced the levels of expression of EGF-R mRNA and TGF-alpha mRNA. In addition, incubation of cells with monoclonal antibody 225 significantly elevated the levels of excreted TGF-alpha species in the culture medium. Our findings suggest that proliferation of human renal carcinoma cells may be regulated by endogenously produced TGF-alpha and that this regulatory pathway can be interrupted using antibody to its receptor, EGF-R.

Topics: Carcinoma, Renal Cell; Cell Division; Epidermal Growth Factor; ErbB Receptors; Humans; RNA, Messenger; Transforming Growth Factor alpha; Tumor Cells, Cultured

The CaKi-I line of renal carcinoma (RC) cells is highly sensitive to the antiproliferative effect of human leukocyte interferon (IFN-alpha). These RC cells express high numbers of cell surface receptors for epidermal growth factor (EGF), and EGF stimulates their proliferation. IFN-alpha blocks EGF-stimulated proliferation of these cells and down-regulates EGF receptors (EGFR) by inhibiting EGFR synthesis. Although EGF stimulates the proliferation of RC cells resistant to the antiproliferative action of IFN-alpha, IFN-alpha treatment does not block the EGF-stimulated proliferation of these cells and has no effect on EGFR expression. Thus, the down-regulation of EGFR is specific for RC cells sensitive to IFN-alpha. While IFN-alpha does not affect the level of total cellular message or total polyadenylated message for the EGFR, IFN-alpha treatment decreases the level of cytoplasmic EGFR message. Analysis of polysome distribution of cellular mRNAs indicates that IFN-alpha treatment results in an accumulation of EGFR mRNA in lighter polysome fractions, consistent with a partial block in translational elongation. Thus, IFN-alpha regulates the expression of EGFR and possibly other growth-related proteins by post-transcriptional mechanisms, which may play an important part in the complex inhibitory action of IFN-alpha on RC proliferation.

Topics: Carcinoma, Renal Cell; Cell Division; Cell Line; Dose-Response Relationship, Drug; Down-Regulation; Epidermal Growth Factor; ErbB Receptors; Gene Expression; Glyceraldehyde-3-Phosphate Dehydrogenases; Humans; Interferon Type I; Kidney Neoplasms; Kinetics; Poly A; Polyribosomes; Recombinant Proteins; RNA; RNA, Messenger

We evaluated the concentrations of immunoreactive epidermal growth factor (EGF) in urine samples from 14 patients with renal cell carcinoma, 16 patients with bladder tumor and 43 non-malignant controls by using radioimmunoassay. In the non-malignant controls, urinary EGF excretion significantly decreased with age (r = -0.46, p less than 0.01), and females excreted significantly more EGF than males [16.3 +/- 7.6 and 9.9 +/- 6.0 (mean +/- SD) ng/mg.creatinine; p less than 0.05]. There was no significant difference between urinary EGF excretion in the patients with renal cell carcinoma and non-malignant controls matched for sex and age (15.9 +/- 12.0 and 14.5 +/- 7.9 ng/mg.creatinine). The difference in excretion of EGF between the patients with bladder tumor and non-malignant controls also was not significant (9.9 +/- 5.6 and 11.7 +/- 7.0 ng/mg.creatinine). These findings indicate, that urine EGF has little usefulness as a tumor marker for renal cell carcinoma and bladder tumor.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Renal Cell; Epidermal Growth Factor; Female; Humans; Kidney Neoplasms; Male; Middle Aged; Radioimmunoassay; Urinary Bladder Neoplasms

Oncogenes and growth factors play a role in normal cellular processes such as growth and differentiation. In addition, considerable circumstantial evidence indicates that these genes may be responsible for the induction and/or maintenance of human malignancies. There are different oncogene families; up to now only a small number has been tested in renal cancer cells.

Topics: Carcinoma, Renal Cell; DNA, Neoplasm; Epidermal Growth Factor; Genes, fos; Genes, myc; Genes, ras; Humans; Kidney Neoplasms; Oncogenes; Tumor Necrosis Factor-alpha

A hereditary form of renal cell carcinoma exists in rats that results from a single gene mutation and is histologically similar to that described in humans. Cell lines derived from these rat tumors were shown to express abundant transforming growth factor-alpha (TGF-alpha) and epidermal growth factor (EGF)-receptor RNA transcripts, but no EGF mRNA. In contrast, normal kidney expressed EGF and EGF-receptor transcripts, but TGF-alpha transcripts were barely detectable. Other kidney epithelial cell lines examined (NRK 52E, MDCK, and LLCPK) were negative for expression of both TGF-alpha and EGF transcripts, but expressed EGF receptors. In addition, the renal cell carcinoma-derived lines secreted TGF-alpha into the media. Immunohistochemistry of normal kidney with a TGF-alpha specific antibody revealed a characteristic pattern of staining of collecting ducts and, to a lesser degree, proximal tubules. In the neoplastic kidney tissue, both the cystic and solid portions of the tumors displayed intense immunoreactivity, indicating that altered expression of this growth factor by the transformed cells occurred in situ. These results suggest that altered TGF-alpha expression is an important aspect of the neoplastic phenotype in rodent as well as human renal cell carcinoma, and support the use of this hereditary rodent tumor model for studying the pathogenesis of this disease.

Topics: Animals; Carcinoma, Renal Cell; Epidermal Growth Factor; ErbB Receptors; Kidney; Kidney Neoplasms; Phenotype; Rats; RNA, Messenger; Transforming Growth Factor alpha

Renal cell carcinomas (RCCs) frequently metastasize to distant organs in their clinical course. However, the mechanism of the metastasis had not been fully elucidated. In vitro invasion assay has been reported to be a rapid method for the evaluation of the invasive potential of various malignant cells. In vitro invasive potential of RCC has not been investigated by this method. Thus, in the present study, we first attempted to characterize the in vitro invasive potential of four human RCC cell lines which had been established in our institute. Secondly, we investigated the influence of two growth factors (EGF, TGF-beta 1) on the invasive potential of these cell lines when the two factors were applied as chemoattractants. SMKT-R-3 and R-4 cell lines showed more cell penetration through Matrigel than SMKT-R-1 and R-2 cell lines, suggesting that the former cell lines have higher invasive potential. While invasive potential varied in each cell line, it was enhanced by EGF in all cell lines. However, TGF-beta 1 suppressed the invasive potential of all four cell lines. These results suggest that two factors have different actions on the invasion of RCCs.

Topics: Carcinoma, Renal Cell; Cell Division; Epidermal Growth Factor; Humans; Kidney Neoplasms; Neoplasm Invasiveness; Transforming Growth Factor beta; Tumor Cells, Cultured

Thirty-two surgical specimens and three cell lines of human gastric cancers were used for subcutaneous transplantation into nude mice, resulting in the establishment of eight (25%) xenografts from the surgical specimens and two (67%) from the cell lines. The localization of epidermal growth factor (EGF) in the surgical specimens and cell lines of the gastric cancers and their xenografts in nude mice was then investigated immunohistochemically. Epidermal growth factor was stained in the cytoplasm of the cancer cells, being detected in 16 (50%) of the 32 surgical specimens and in all of the cell lines. Seven (44%) of the sixteen EGF-positive surgical specimens and one (6%) of the 16 EGF-negative ones were tumorigenic in nude mice. All of the xenografts in nude mice were positive for EGF. The tumorigenicity of human gastric cancer xenografts in nude mice may, therefore, be correlated with the presence of EGF in cancer cells.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Animals; Carcinoma, Renal Cell; Epidermal Growth Factor; Female; Humans; Kidney Neoplasms; Male; Mice; Mice, Nude; Middle Aged; Neoplasm Metastasis; Neoplasm Transplantation; Staining and Labeling; Stomach Neoplasms; Tumor Cells, Cultured; Wilms Tumor

We have analyzed the expression of the genes for the precursors of epidermal growth factor (pro-EGF) and transforming growth factor alpha (proTGF-alpha) as well as for the EGF receptor in tissue specimens of a large number of adult patients with renal cell carcinoma. Since normal kidney tissue was available from the same patients we could directly compare the expression of these genes in tumors with that in adjacent normal renal tissue. Our experiments reveal underexpression of the proEGF gene in all tumors analyzed (21 of 21) and overexpression of the genes for proTGF-alpha (33 of 33 analyzed) and EGF receptor (22 of 23 analyzed) in tumor samples, when compared with normal kidney tissue. The expression of the proTGF-alpha gene appeared to depend on grade and differentiation of the tumor, since well differentiated tumors (grade 1) expressed more proTGF-alpha mRNA than the adjacent normal tissue but significantly less than poorly differentiated tumors (grade 2 or 3), which are the most aggressive ones. In none of these tissue specimens did we find, by Southern analysis, amplification of the proTGF-alpha or EGF receptor gene. Therefore, overexpression of these genes must be due to another effect, perhaps an alteration of their mRNA turnover. Although the EGF receptor gene (c-erbB1) is overexpressed in nearly all carcinomas analyzed, there was no linear coexpression with the proTGF-alpha gene. In contrast, transcription of the proEGF gene was completely turned off in tumor tissue. Although we have found by restriction fragment length polymorphism analysis, in one of three tumor samples, evidence for a somatic mutation within the proEGF gene, we do not know yet, due to the limited number of Southern analyses, whether this somatic mutation is causally involved in the decrease of proEGF mRNA expression and, hence, is representative of renal cell carcinoma. To our knowledge, this is the first observation on primary tumor tissue in humans that upon malignant transformation the gene for a polypeptide growth factor gene is underexpressed.

Topics: Blotting, Northern; Blotting, Southern; Carcinoma, Renal Cell; DNA, Neoplasm; Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation, Neoplastic; Gene Rearrangement; Humans; Kidney; Kidney Neoplasms; Protein Precursors; RNA, Messenger; Transforming Growth Factors

Epidermal growth factor receptor (EGF-R) was estimated in Hypernephroma by saturation analysis using 125-I EGF as ligand. Tissue levels of this oncogene related protein were increased five-fold (24 to 99 fmol/mg protein) in comparison with the surrounding tumor free tissue (3 to 18 fmoles/mg protein). There was no apparent correlation to the stage of tumor growth or tumor differentiation and there was no correlation of EGF serum levels with tumor growth. EGF serum levels in the tumor patients did not exceed levels in control patients.

Topics: Adult; Aged; Carcinoma, Renal Cell; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Kidney Neoplasms; Male; Middle Aged; Tumor Necrosis Factor-alpha

Tissue extracts prepared from human renal tissue, renal cell carcinoma and serum-free conditioned media of ACHN cells and A498 cells, cell line originated from human renal cell carcinoma, stimulated DNA synthesis of BALB/c 3T3 cells. The activity (growth factor activity) was significantly higher in renal cell carcinoma than in normal tissues. Radioreceptor assay revealed that the contents of epidermal growth factor and type alpha transforming growth factor in the tissue extracts from renal cell carcinoma and conditioned media from renal cell carcinoma cell lines were below detectable level. Most of growth factor activity of the tissue extracts and conditioned media showed high affinity for heparin-Ultrogel, indicating that the major growth factor activity was due to heparin-binding growth factor(s). In addition, renal cell carcinoma contained growth factor activity for ACHN cells, which did not show specific affinity for heparin-Ultrogel.

Topics: Binding, Competitive; Carcinoma; Carcinoma, Renal Cell; Chromatography, Affinity; DNA; Epidermal Growth Factor; ErbB Receptors; Growth Substances; Heparin; Humans; Kidney Neoplasms; Tumor Cells, Cultured

In cells derived from two human renal carcinomas only the precursor form of epidermal growth factor (EGF) was found. The binding assay revealed a high level of EGF receptor expression in both cell types tested. However, these receptors are not involved in the growth activity of the cells under in vitro conditions used. The source of DNA synthesis-stimulating activity found in conditioned media of the cells tested is discussed with respect to possible participation of TGF beta.

Topics: Carcinoma, Renal Cell; Cell Division; Cell Line; Epidermal Growth Factor; ErbB Receptors; Humans; Kidney Neoplasms; Kinetics; Suramin