Effect Of TRPV5 And TRPV6 On The Biological Behavior Of Colon Cancer Cell Line SW480

Objective: In the TRP superfamily, TRPV5 and TRPV6 are the only known calcium ion channels with high selectivity.The present study through observation on the protein expressions of TRPV5 and TRPV6 channels in SW480 colon cancer cell, and by changing the functional status of TRPV5 and TRPV6 channels to observe the Ca2+concentration in SW480 colon cancer cell as well as the regulating effect of SW480 cell on growth and apoptosis. The purpose of this study is focusing on the effect of TRPV5 and TRPV6 on biological behavior of SW480 colon cancer cell.so as to provide theoretical and experimental basis for diagnosis and treatment of TRPV5 and TRPV6 in colon cancer in further study.Methods:(1) The real-time fluorescence quantitative PCR technology, Western-Bolt technology and cell immunohistochemical technology are adopted to detect the comparison of TRPV5 and TRPV6 agonists 1,25(OH)2D3(10-8mol/l) and Ca Cl2(10mmol/l) and the inhibitor Cu Cl2(100μmol/l) on TRPV5 m RNA, TRPV6 m RNA and protein level expression in SW480 colon cancer cell.(2) After changing the functional status of TRPV5 and TRPV6 channels by agonists and inhibitors, the imaging system of high-speed iron is appied to observe the effect of Ca2+concentration change of TRPV5 and TRPV6 in SW480 colon cancer cell;(3) After the synchronized treatment on colon cancer SW480 cell, using the cell proliferation assay(MTT colorimetry), cell migration assay(scratch repair method) and cell apoptosis assay(biochemical detection Tunel method) to observe the effect of agonists 1,25(OH)2D3 and Ca Cl2 as well as inhibitor Cu Cl2 on the proliferation, migration and apoptosis of SW480 colon cancer cell.Results:(1) By comparing with the control group, TRPV5 mRNA, TRPV6 m RNA and protein content in the agonist 1,25(OH)2D3 and Ca Cl2 group are significantly increased(P<0.05); while in inhibitor Cu Cl2 group, the expressions are significantly decreased(P<0.05). There is no significant difference in agonist 1,25(OH)2D3 and Ca Cl2 groups(P>0.05); by comparing the agonist 1,25(OH)2D3 and Ca Cl2 groups with inhibitor Cu Cl2 group, the TRPV5 m RNA, TRPV6 m RNA and protein content are significantly increased(P<0.01);(2) Under the action of agonist1,25(OH)2D3, the concentration of Ca2+is increased in SW480 colon cancer cell compares with the control group(P<0.05); while under the action of inhibitor Cu Cl2, the concentration of Ca2+ is decreased in SW480 colon cancer cell compares with the control group(P<0.05);(3) The agonist 1,25(OH)2D3 and Ca Cl2 have the effect of promoting proliferation, migration, and inhibiting apoptosis on SW480 colon cancer cell(P<0.05), with dependence of concentration and time; while the inhibitor Cu Cl2 has the effect of inhibiting proliferation, migration, and promoting apoptosis on SW480 colon cancer cell(P<0.05), with dependence of concentration and time.Conclusion:(1)The agonists 1,25(OH)2D3 and Ca Cl2 can make TRPV5 m RNA, TRPV6 m RNA and protein increase expression in SW480 colon cancer cell. It make Ca2+concentration increase in SW480 colon cancer cells, promoting proliferation and migration and inhibition of apoptosis on SW480 colon cancer cell.(2)The Inhibitors Cu Cl2 can make TRPV5 m RNA,TRPV6 m RNA and protein reduce expression in SW480 colon cancer cell. It make Ca2+concentration reduce in SW480 colon cancer cells, inhibitiing proliferation and migration and induction of apoptosis on SW480 colon cancer cell.(3)TRPV5 and TRPV6 are expressed in SW480 colon cancer cell, but with TRPV6 more apparent.Changing TRPV5 and TRPV6 channel protein expression and by increasing or decreasing the content of intracellular Ca2+, further adjust the biological behavior of SW480 colon cancer cell.