| Background:Gastric cancer is the fifth most commen malignant tumor in the world.TRPV6 is a new type of calcium pathway,which induces the proliferation of tumor cells by activating the ERK pathway.?-lapraquinone is a popular drug for the treatment of gastric cancer,which can inhibit the proliferation and migration of tumor cells by inhibiting ERK/mTOR/ART and other pathways.So is there a certain relationship between ?-lapachone and TRPV6 in gastric cancer cell line,and what is the mechanism is not clear yet.Objective:To investigate the biological effects of ?-lapachone on SGC-7901 gastric cancer cell line with overexpression of TRPV6.Analysis of its inhibitory mechanism provides a clinical basis for the future targeted therapy of gastric cancer.Method:The expression of TRPV6 in gastric cancer cell line AGS and SGC-7901 was detected by Western blot technique,and the gastric cancer cell line expressing TRPV6 was found out in SGC-7901.The expression of TRPV6 protein was reduced by gene transfection and silencing.The proliferative ability of gastric cancer cells was detected by MTT assay in 4 groups:gastric cancer with ?-lapachone group,gastric cancer group,?-lapachone with TRPV6 transfection group and TRPV6 transfection group(P<0.05).The migration ability of gastric cancer cells in four groups was detected by scratch test.Western blot was used to detect the changes of cell cycle related protein expression in the four groups.The above experiments were repeated three times and compared with each other.Result:1.Western blot showed that TRPV6 only existed in SGC-7901 gastric cancer cell lines.in AGS without TRPV6 expression.transfection results showed that the 50nm and 100nm concentration of the si-RNA sequence of 2 can silent TRPV6,and 100nm silencing can reach more than 75%.3.After transfection of TRPV6,Western Blot result indicating the TRPV6 were successful transfected.After successful transfection,using MTT,migration test to test four group cells'ability of proliferation and migration.siRNA-TRPV6 ? with lapachone tumor cells in the four groups was the weakest,and the proliferation and migration ability of the tumor cells in ?-lapraquinone group and si-TRPV6 group was also weakened,and the degree was almost the same.4.The expression of ERK was significantly inhibited and the expression of ERK was the least in the four groups.The expression of ERK in ?-lapachone group and si-TRPV6 group was down-regulated,and the degree was similar.The results showed that ?-lapachone inhibited.The mechanism of ?-lapachone inhibitor TRPVP6 overexpression in SGC-7901 gastric cancer cell line is as follows:1.?-Lapachone can produce oxygen free radical and then NAD/ATP depletion,while the transport of calcium ion requires energy depletion and the transport of TRPV6 is inhibited.In turn,the proliferation and migration of tumor cells were inhibited,leading to apoptosis of tumor cells.2.?-Lapachone also inhibited the ERK pathway,and the inhibition of EMT.ERK pathway in tumor cells was the main mechanism by which TRPV6 induced abnormal proliferation and migration of tumor cells.?-Lapraquinone inhibited TRPV6 to promote tumor cell proliferation.Colonization and migration of the main pathway,resulting in apoptosis of tumor cells.Conclusion:Inhibition of TRPV6-overexpressing SGC-7901 gastric cancer cell lines by ?-lapachone is mainly through inhibiting the ERK pathway,reducing the formation of ATP in gastric cancer cells,and inhibiting the abnormally metabolized calcium transporter cancer cells,resulting in cancer cells' proliferation and migration decreased,eventually leading to apoptosis in gastric cancer cells. |
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