The Effects Of TRPV6 On The Occurrence Of Colon Cancer

Objective:Previous experimental studies found that the regulation of TRPV6 protein and mRNA expression by corresponding agonists and inhibitors can make the Ca²⁺level of colon cancer SW480 cells change accordingly,thus affecting the proliferation,migration and inhibition of apoptosis of colon cancer SW480 cells.By observing the effect of TRPV6 gene silencing on the calcium and biological behaviors of colon cancer SW480cells and the effect of TRPV6 on the occurrence and development of colonic tumors in SD rats.To further explore the TRPV6 in the occurrence and development of colon cancer mechanism and its role in the prevention and treatment of colon cancer looking for new mechanisms and therapeutic targets.Methods:?1?Effect of TRPV6 gene silencing on calcium and biological behavior of colon cancer SW480 cells:The TRPV6 interference sequence was designed by the Shanghai genechem company,fragment synthesis after construction of lentiviral RNAi expression vector,then packaged into mature lentivirus to infect SW480 cells by puromycin selection after cultured SW480 cell lines stably expressed TRPV6–RNAi.Through Western-Bolt and PCR technology and immunohistochemical technique to verify the expression of the transfected successfully and effectively inhibit TRPV6 protein and mRNA,and then MTT colorimetric assay,cell migration assay and cell apoptosis assay were observed after TRPV6 gene silencing on the proliferation,migration and apoptosis of colon cancer SW480 cells and the effect of SW480 cells Ca²⁺concentration change.?2?Effects of TRPV6 on the development and progression of colonic tumors in SD rats:Fifty-five SD rats were selected and divided into five groups randomly,ten in each intervention group which were the DMH+1,25?OH?₂D₃ group and DMH+CaCl₂ group and DMH+CuCl₂ group,15 in the DMH group and 10 in the control group.DMH+1,25?OH?₂D₃ group was given intraperitoneal injection of dimethylhydrazine?20mg/kg?once a week and fed with 1,25?OH?₂D₃ 37.5nmol/kg every other day;DMH+CaCl₂ roup was given intraperitoneal injection of dimethylhydrazine?20mg/kg?once a week and fed with CaCl₂ 37.5mmol/kg every other day;DMH+CuCl₂ group was given intraperitoneal injection of dimethylhydrazine?20mg/kg?once a week and fed with CuCl₂ 375umol/kg every other day;DMH group was only given intraperitoneal injection of dimethylhydrazine?20mg/kg?once a week and fed with the same amount of saline every other day;The control group was given intraperitoneal injection of equal amount of normal saline every week and fed with the same amount of saline every other day.In 18weeks,20 weeks,22 weeks killed a group of mice.The construction of the model was detected by HE staining.The expression of TRPV6 protein in large intestine was detected by Western-Blot blot.Results:?1?The TRPV6-RNAi group was able to reduce the concentration of Ca²⁺in colon cancer SW480 cells,and compared with the two control group,the difference was statistically significant?P<0.05?.?2?The TRPV6-RNAi group could inhibit the proliferation and migration of colon cancer SW480 cells and reduced its apoptosis,and compared with the two control group,the difference was statistically significant?P<0.05?.?3?In the control group,no tumor was found in the large intestine,but 25 of the 37rats in the experimental group and the intervention group were successfully induced by the large intestine tumor,the rate of tumor formation was 67.6%,the tumorigenicity rate of DMH+1,25?OH?2D3 group was 100%,the DMH group was 84.62%,the DMH+CaCl₂group was 50%,and the DMH+CuCl₂ group was 33.33%.?4?The expression of TRPV6protein was detected by Western Blot blot in all groups of large intestine tissues,the expression of TRPV6 protein in DMH group,DMH+1,25?OH?₂ D₃ group,DMH+CaCl₂group and DMH+CuCl₂ group were all higher than those in the control group,and the difference was statistically significant?P<0.05?.The expression of TRPV6 protein in DMH group was higher than in DMH+CaCl₂ group and DMH+CuCl₂ group,and the difference was statistically significant?P<0.05?,while the difference between DMH+CaCl₂ group and DMH+CuCl₂ group was not statistically significant?P>0.05?.The expression of TRPV6protein in DMH+1,25?OH?₂D₃ group was higher than in the DMH group,and the difference was statistically significant?P<0.05?.Conclusion:?1?After transfection of TRPV6-RNAi to colon cancer SW480 cells,the concentration of Ca²⁺in SW480 cells of colon cancer is reduced,and can inhibit the proliferation,migration and promote its apoptosis of colon cancer SW480 cells.?2?1,25?OH?₂D₃ can increase the expression of TRPV6 protein in the colon tissues of the rats and promote the formation of colonic tumor.?3?CaCl₂ and CuCl₂ can reduce the expression of TRPV6 protein in the colon tissues and inhibit the formation of colonic tumor.