Effect of TRPV5 and TRPV6 channels on the biological behaviors of SW480 colon cancer cell line / 中国肿瘤临床

ABSTRACT

Objective:To investigate the role of TRPV5 and TRPV6 in intracellular calcium regulation and biological behaviors of SW480 colon cancer cells. Methods:qRT-PCR, Western blot, and immunocytochemistry were applied to determine the mRNA and protein ex-pression levels of TRPV5 and TRPV6 in SW480 colon cancer cell line upon treatment with TRPV5 and TRPV6 agonist, 1-25(OH)2D3, and inhibitor, CuCl2. The change of intracellular Ca2+level was examined with a confocal laser scanning microscope. Scratch test, MTT, and TUNEL assays were used to analyze the cell migration, proliferation, and apoptosis, respectively. Results:As an agonist of TRPV5 and TRPV6, 1-25(OH)2D3 significantly up-regulated the mRNA and protein expression levels of TRPV5 and TRPV6 in SW480 cell lines. On the other hand, CuCl2, being an inhibitor of TRPV5 and TRPV6, effectively down-regulated the TRPV5 and TRPV6 mRNA and protein expres-sion levels (P<0.05). The intracellular calcium concentration in SW480 cell line significantly increased upon treatment with 1-25 (OH)2D3, and significantly decreased with CuCl2 treatment (P<0.05). 1-25(OH)2D3 promoted cell proliferation and migration, and inhibit-ed apoptosis of SW480 cell in a time-and dose-dependent manner (P<0.05). However, CuCl2 significantly repressed cell proliferation and migration and induced apoptosis (P<0.05). Conclusion: TRPV5 and TRPV6 can affect the biological behaviors of colon cancer SW480 cells by regulating intracellular Ca2+level.