cholecystokinin and Melanoma

To explore the feasibility of RNA interference in the treatment of melanoma by inhibiting the Foxp3 gene expression in mouse B16 melanoma cells using RNA interference (RNAi) in vitro.. Small interfering RNA (siRNA) was designed according to Foxp3 gene. A short hairpin RNA (shRNA) lentivirus expression vector was constructed and transfected into mouse B16 cells, and RNA interference was induced in vitro. Western blot and real-time RT-PCR were performed to detect the expression of Foxp3 gene. ELISA was applied to detect the changes of TGF-β(1);, TGF-β(2);, IL-10 and other cytokines. The B16 cells after interference were co-cultured with CD4(+);CD25(-);T lymphocytes. CCK8 assay was used to monitor the proliferation of CD4(+);CD25(-);T lymphocytes.. shRNA could suppress the expression level of Foxp3, down-regulate the inhibitory ability of tumor cells on the proliferation of CD4(+);CD25(-);T lymphocytes, and reduce the secretion of TGF-β(1);, TGF-β(2);, IL-10 and other cytokines, in particular the expression of TGF-β(2);.. RNA interference can inhibit the expression of target gene Foxp3 in mice melanoma cells and the proliferation of tumor cells. It can also reduce the inhibition on the proliferation of CD4(+);CD25(-);T lymphocytes, and the secretion of inhibitory cytokines.

Topics: Animals; Blotting, Western; CD4-Positive T-Lymphocytes; Cell Line, Tumor; Cell Proliferation; Cells, Cultured; Cholecystokinin; Coculture Techniques; Forkhead Transcription Factors; Gene Expression Regulation, Neoplastic; Green Fluorescent Proteins; Interleukin-10; Lentivirus; Melanoma; Mice; Microscopy, Fluorescence; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; RNA, Small Interfering; Transfection; Transforming Growth Factor beta1; Transforming Growth Factor beta2

The effects of gastrin (G17) on the growth and migration factors of four human melanoma cell lines (HT-144, C32, G-361, and SKMEL-28) were investigated. The expression patterns of cholecystokinin (CCK)(A), CCK(B), and CCK(C) gastrin receptors were investigated in these cells and in seven clinical samples by means of reverse transcription polymerase chain reaction. Melanoma cells appear to express mRNA for CCK(C) receptors, but not for CCK(A) or CCK(B) receptors. Although gastrin does not significantly modify the growth characteristics of the cell lines under study, it significantly modifies their cell migration characteristics. These modifications occur at adhesion level by modifying the expression levels of alpha(v) and beta3 integrins, at motility level by modifying the organization of the actin cytoskeleton, and at invasion level by modifying the expression levels of matrix metalloproteinase 14. We recently demonstrated the presence of CCK(B) receptors in mouse endothelial cells involved in glioblastoma neoangiogenesis. Chronic in vivo administration of a selective CCK(B) receptor antagonist to mice bearing xenografts of human C32 melanoma cells significantly decreased levels of neoangiogenesis, resulting in considerable delays in the growth of these C32 xenografts. In conclusion, our study identifies the pleiotropic effects of gastrin on melanoma cell biology.

Topics: Actins; Animals; Antineoplastic Agents; Apoptosis; Blotting, Western; Cell Line, Tumor; Cell Movement; Cholecystokinin; Cisplatin; Cytoskeleton; Dacarbazine; DNA Primers; DNA, Complementary; Flow Cytometry; Gastrins; Gene Expression Regulation, Neoplastic; Glioblastoma; Humans; Immunohistochemistry; Melanoma; Mice; Neoplasm Invasiveness; Neoplasm Transplantation; Neovascularization, Pathologic; Oligonucleotide Array Sequence Analysis; Poly(ADP-ribose) Polymerases; Receptor, Cholecystokinin B; Receptors, Cholecystokinin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Time Factors; Wound Healing

Topics: Amyloidosis; Biochemical Phenomena; Biochemistry; Cholangitis; Cholecystitis; Cholecystokinin; Colonic Neoplasms; Drug Therapy; Duodenum; Electrophoresis; Enteritis; Gastritis; Hemosiderosis; Humans; Intestinal Secretions; Leucyl Aminopeptidase; Liver Cirrhosis; Liver Diseases; Melanoma; Mucous Membrane; Nephrosis; Pancreatitis