1.Effect of TRPV6 channels on the biological behavior of sw480 colon cancer cell line and colonic tumorigenesis in rat model
Chenwu XIE ; Morong LIU ; Hui DONG ; Jingyu XU ; Rui XIE ; Guorong WEN ; Lianhua LIU
Chinese Journal of Clinical Oncology 2018;45(21):1079-1085
Objective: To observe the influence of TRPV6 gene silence on SW480 colon cancer cell biological behavior, change in the in-tracellular concentration of calcium, as well as influence of 1,25 (OH)2D3CaCl2and CuCl2on SD rat colonic neoplasm models. Method: SW480 colon cancer cells were infected using lentivirus particles. TRPV6 protein and mRNA expression was detected using immunohistochemical tests, Western blot, and PCR. Moreover, the proliferation, metastasis, and apoptosis of SW480 colon cancer cells were detected through MTT assay and metastasis and apoptosis experiments, and the concentration of Ca2+in SW480 colon cancer cells was measured using high-speed ionic imaging. The SD rat colon cancer model was established based on DMH, and were assigned into experimental group (DMH group, 15) and intervention group (DMH+1,25 (OH)2D3group, DMH+CuCl2group) and control group, 10 in each group. The SD rat colon cancer model is established based on DMH, given 1,25(OH)2D3(37.5 nmol/kg) and CuCl2(375 μmol/kg) separately as intervention. The occurrence of colonic neoplasms and glandular cancers in each group of rats was observed, and Western blot was employed for detection of the TRPV6 protein expression. Results: After the transfection of SW480 colon cancer cells by TRPV6-RNAi, the expression of TRPV6 mRNA and protein decreased, intracellular concentration of Ca2+decreased, proliferation and metastasis rate of SW480 colon cancer cells decreased, and apoptosis rate of these cells increased. The differences between the groups with intervention and the blank control group and negative control group showed statistical significance (P<0.05). The colon cancer occurrence rate in the control group was 0, while that of the DMH+1,25 (OH)2D3 group, DMH group, and DMH+CuCl2were 100%, 84.62%, and 33.33%, respectively. The TRPV6 protein expression was detected in all groups, while DMH+1,25(OH)2D3group was observed to exhibit the highest level of expression, followed by the DMH group, DMH+CuCl2group, and control group. The differences were of statistical significance (P<0.05). Conclusions: The proliferation and metastasis of SW480 colon cancer cells can be prohibited by lowering the concentration of Ca2+in the cells. Thus, the apoptosis of the cells can be induced. 1,25 (OH)2D3can help improve the expression of TRPV6 protein in experimental rat colon tissues and promote the formation of colon neoplasms. CuCl2can help lower the expression of TRPV6 protein in experimental rat colon tissues and prevent the formation of colon neoplasms.