sincalide and Carcinoma

Kidney renal clear cell carcinoma (KIRC) is the most common pathological subtype of kidney tumor. Reportedly, LINC00174 is a key regulator in cancer progression. This study aims to clarify the role and molecular mechanism of LINC00174 in the progression of KIRC.. LINC00174 expression in KIRC and its prognostic value were analyzed by bioinformatics. LINC00174, miR-612 and FOXM1 mRNA expression levels in KIRC clinical samples and cell lines were detected by qRT-PCR. After LINC00174 was overexpressed or knocked down, CCK-8, BrdU and Transwell assays were adopted to evaluate the proliferation and metastatic potential of KIRC cells. Bioinformatics and dual luciferase reporter assays were employed to validate the targeting relationship between miR-612 and LINC00174 or FOXM1 mRNA, respectively. Western blot assay was performed to detect FOXM1 protein expression in KIRC cells.. LINC00174 expression and FOXM1 expression were up-regulated in 42 cases of KIRC tissues (

Topics: Bromodeoxyuridine; Carcinoma; Cell Line, Tumor; Cell Movement; Cell Proliferation; Forkhead Box Protein M1; Gene Expression Regulation, Neoplastic; Humans; MicroRNAs; RNA, Messenger; Sincalide

The high risk of recurrence faced by patients with bladder cancer has necessitated the administration of supplemental intravesical chemotherapy; however, such treatments often result in severe side effects. As a result, novel intravesical agents with enhanced efficacy and minimal toxicity are urgently required for the treatment of bladder cancer.. Guizhi Fuling Wan (GFW) is a traditional Chinese medicine shown to inhibit the growth of hepatocellular carcinoma. This study evaluated the growth inhibition of GFW using normal human urothelial cells and bladder cancer cells; the efficacy of GFW treatment was further compared with mitomycin C, epirubicin, and cisplatin. We also examined the progression of cell cycle and apoptosis in bladder cancer cells in response to GFW treatment. CCK-8 was employed to analyze cell viability and flow cytometry was used to study the cell cycle and apoptosis. The mechanisms underlying GFW-induced cell cycle arrest were determined by Western blot analysis.. Our data demonstrate the potent inhibitory effect of GFW in the proliferation of bladder cancer cell lines, BFTC 905 and TSGH 8301. GFW presented relatively high selectivity with regard to cancer cells and minimal toxicity to normal urothelial cells. Our results also demonstrate that GFW interferes with cell cycle progression through the activation of CHK2 and P21 and induces apoptosis in these bladder cancer cells.. Our results provide experimental evidence to support GFW as a strong candidate for intravesicle chemotherapy against bladder cancer.

Topics: Antineoplastic Agents; Antineoplastic Agents, Phytogenic; Apoptosis; Carcinoma; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Checkpoint Kinase 2; Cisplatin; Cyclin-Dependent Kinase Inhibitor p21; Drugs, Chinese Herbal; Epirubicin; Humans; Mitomycin; Phytotherapy; Protein Serine-Threonine Kinases; Sincalide; Urinary Bladder; Urinary Bladder Neoplasms; Urothelium

To study the effect of microenvironment simulated by esophageal carcinoma homogenate supernatant on the differentiation and development of human dendritic cells (DCs) and to investigate the mechanisms of tumor immune escape for the clinical application of DC vaccines.. Fresh esophageal carcinoma and peri-cancer tissues were collected to prepare homogenate supernatant and the content of VEGF-A was detected by ELISA. The peripheral blood monouclear cells were isolated by density gradient centrifugation and cultured with RPMI1640 medium including rhGM-CSF and rhIL-4 to induce to DCs. Then the esophageal carcinoma homogenate supernatant, peri-carcinoma homogenate supernatant and VEGF-A were added on the second day and half of the medium was changed every other day. Antigen of esophageal carcinoma cell line EC9706 was added on day 4 and lipopolysaccharide (LPS) was added on day 6. DCs were collected on day 8 for further study. Checked the morphology of DCs by microscope, the immunophenotype by flow cytometry, the gene of CD1a by RT-PCR and the proliferation and killing rate of T cell by CCK-8.. The content of VEGF-A in the homogenate supernatant of esophageal carcinoma was significantly higher than that of the peri-carcinoma (0.987+/-0.319 microg/L, 0.152+/-0.105 microg/L, P<0.05). The cell morphology in esophageal carcinoma homogenate supernatant group was inhibited. Besides, compared with normal DCs, the positive expression rate of CD86 decreased from 69+/-8 to 42+/-11, CD1a decreased from 56+/-12 to 27+/-12 and CD11c decreased from 21+/-13 to 18+/-13 (P<0.01). CD1a gene almost showed no expression. The proliferation capacity of T cells decreased from 112.53+/-7.16 to 70.18+/-3.47 (P<0.01), and their killing capacity of T cells decreased from 62.42+/-0.57 to 46.81+/-1.62 (P<0.01). However, the cells had no difference among peri-carcinoma homogenate supernatant group, VEGF-A group and normal DC group.. The tumour microenvironment stimulated by the esophageal carcinoma homogenate supernatant obviously has inhibitory effect on the differentiation and function of DCs.VEGF-A may not be the key factor in the process.

Topics: Antigens, CD1; B7-2 Antigen; Carcinoma; Cell Differentiation; Cell Line, Tumor; Cells, Cultured; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Esophageal Neoplasms; Flow Cytometry; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunophenotyping; Interleukin-4; Lipopolysaccharides; Reverse Transcriptase Polymerase Chain Reaction; Sincalide; T-Lymphocytes; Vascular Endothelial Growth Factor A

Pro-cholecystokinin (CCK) has three sulfated tyrosine residues. Sulfation of the tyrosine residue in CCK 8 is known to be important for its activity at CCK A receptors. The role of these sulfated tyrosines in the sorting and processing of pro-CCK was examined by treatment of CCK-secreting rat thyroid medullary carcinoma cells with 10 nM sodium chlorate (a non-toxic inhibitor of tyrosine sulfation). This treatment caused a 50% decrease in the cellular content of immunoreactive CCK and an 80% decrease in its secretion. Sephadex G-50 chromatography of cellular extracts and culture media showed a selective depletion of CCK 8. There was a comparative sparing of CCK 33 and larger molecular forms in cellular extracts which was not observed in the media. These results suggest that the sulfation of the tyrosines of pro-CCK is clearly important for the correct sorting and/or processing of pro-CCK. The pattern of immunoreactive CCK peptides seen with chlorate treatment is consistent with the substrate specificity of a recently identified putative CCK cleaving enzyme and suggests that unsulfated pro-CCK is not efficiently processed to CCK 8 in vivo. The large decrease in CCK content and secretion observed with sodium chlorate may also be due to inefficient sorting of unsulfated pro-CCK into secretory vesicles.

Topics: Animals; Carcinoma; Chlorates; Cholecystokinin; Culture Media; Protein Precursors; Rats; Receptors, Cholecystokinin; Sincalide; Sulfates; Sulfur Radioisotopes; Thyroid Neoplasms; Tumor Cells, Cultured; Tyrosine

Gastrointestinal hormones and neuropeptides are known to regulate growth of various normal gastrointestinal tissues and many cancers. Since cholecystokinin (CCK) is considered the most potent trophic factor for the exocrine pancreas, we studied its effect on growth of an acinar cell tumor, initially induced by azaserine and transplanted to the rat, in comparison with the normal pancreas. When tumors became palpable, rats were treated three times daily for 12 or 14 days with CCK-8 or NaCl 0.9% (controls) alone or in combination with the CCK receptor antagonist CR1409 (10 mg/kg) administered subcutaneously twice daily. Then tumors and pancreata were analyzed for their size, composition, and CCK receptors. Tumor volume, weight, and protein content, RNA, DNA, and enzymes decreased after CCK-8 treatment in a dose-dependent manner, the maximal effect being observed with 4-micrograms/kg treatment. This inhibitory effect was partially suppressed by CR1409, which by itself also reduced tumor growth, but to a lesser degree. CCK-8 exerted a stimulating effect on growth of the normal pancreas with low doses (1 and 2 micrograms/kg) and an inhibitory effect or no effect with a higher dose (4 micrograms/kg). CR1409 prevented this latter effect, but did not affect by itself the normal pancreas. These findings suggest that CCK-8 inhibits growth of an acinar cell tumor grafted to the rat; this effect is mediated by the occupation of specific CCK receptors present in high density on these cells. In contrast, CCK-8 exerts a biphasic effect on the normal pancreas as a function of its dose.

Topics: Animals; Carcinoma; Cell Division; Dose-Response Relationship, Drug; Neoplasm Transplantation; Pancreatic Neoplasms; Proglumide; Rats; Rats, Inbred Lew; Receptors, Cholecystokinin; Sincalide

A rat medullary thyroid carcinoma cell line, CA-77, was shown to express the cholecystokinin (CCK) gene. Measurements using a library of sequence-specific radioimmunoassays before and after enzymic treatment of extracts and chromatographic fractions showed that the cells contained 1.0 pmol of alpha-carboxyamidated cholecystokinins/10(6) cells, 0.4 pmol of glycine-extended intermediates/10(6) cells and 1.0 pmol of further C-terminal-extended pro-CCK/10(6) cells. Gel chromatography and reverse-phase h.p.l.c. revealed both sulphated and nonsulphated CCK-8 in the cells. The growth medium contained in addition alpha-amidated CCK-33, glycine-extended CCK-8 and pro-CCK. Exposure to 0.1 microM-dexamethasone for 6 days increased the cellular content and secretion of all of the described CCK peptides by 2-3-fold. The increase was first noted after 3 days of treatment. Monensin inhibited the synthesis of alpha-carboxyamidated CCK and the secretion of all of the CCK forms measured. Colchicine at a low concentration (0.2 mumol/l) apparently increased the synthesis and secretion of alpha-carboxyamidated CCK, whereas higher concentrations inhibited CCK synthesis. Finally, chloroquine inhibited the alpha-carboxyamidation of CCK. We conclude that the CA-77 cell line is a useful tool for studies of the expression and post-translational processing of pro-CCK.

Topics: Animals; Carcinoma; Chloroquine; Cholecystokinin; Chromatography, Gel; Colchicine; Dexamethasone; Gene Expression; Glycine; Monensin; Peptide Fragments; Protein Precursors; Protein Processing, Post-Translational; Rats; Sincalide; Sulfates; Thyroid Neoplasms; Tumor Cells, Cultured

The secretion of protein, like cell proliferation, is an integrated response that reflects structural organization of the cell periphery. Stimulation of protein secretion was thus utilized for comparison of integrated responses of the cell periphery in pancreatic acinar carcinoma of the rat and integrated responses in normal rat pancreas. Results of this comparison include: a) The stimulation of protein secretion in acinar carcinoma fragments by carbamylcholine chloride and cholecystokinin octapeptide, pancreatic secretagogues that interact with specific plasma membrane receptors, was only a fraction (one-fifth to one-half) of that observed in normal pancreatic minilobules. b) The Ca2+ ionophore A23187 and the cyclic nucleotide N6,O2'-dibutyryl cyclic AMP, secretagogues that act independently of specific membrane receptors, did not stimulate secretion in the acinar carcinoma. The observed quantitative and qualitative differences in protein secretion indicate fundamental differences in cell periphery organization between the normal and transformed acinar cells of pancreas.

Topics: Animals; Bucladesine; Calcimycin; Carbachol; Carcinoma; Cell Membrane; Cholecystokinin; Dose-Response Relationship, Drug; In Vitro Techniques; Pancreas; Pancreatic Neoplasms; Peptide Fragments; Proteins; Rats; Secretory Rate; Sincalide; Temperature