七氟烷联合下调CTRP6基因抑制肝癌细胞迁移和侵袭的机制研究

Mechanism of Sevoflurane Combined with Down-Regulation of CTRP6 Gene to Inhibit Migration and Invasion of Hepatoma Cells

目的探讨七氟烷联合下调CTRP6对肝癌细胞迁移侵袭的影响及作用机制。方法运用MTT法检测七氟烷(1%、2%、5%)对肝癌细胞的抑制率;将si-con组(转染si-con)、si-CTRP6(转染si-CTRP6)、sevo+si-con(转染si-con,再用2%sevo处理)、sevo+si-CTRP6组(转染si-CTRP6,再用2%sevo处理),用脂质体法转染至HepG2;qRT-PCR法检测细胞中CTRP6的表达;Transwell法检测细胞的迁移和侵袭;Western blot检测细胞中CTRP6、Shh、Ihh、Smo的蛋白表达。结果与control组相比,2%sevo、5%sevo可明显增强肝癌细胞的抑制率,且2%sevo可明显的抑制肝癌细胞的迁移和侵袭(P<0.05);重要的是,2%sevo可下调肝癌细胞中CTRP6的表达,且敲减CTRP6可明显的抑制肝癌细胞的迁移和侵袭;敲减CTRP6可明显的抑制肝癌细胞中Hedgehog信号通路关键基因Shh、Ihh、Smo的蛋白表达。结论七氟烷联合下调CTRP6可抑制肝癌细胞的迁移和侵袭,其机制与失活Hedgehog信号通路有关,将可为七氟烷联合下调CTRP6在肝癌中的治疗提供实验支持。

Objective To investigate the effect of sevoflurane combined with down-regulation of CTRP6 on migration and invasion of hepatoma cells and its mechanism.Methods MTT assay was used to detect the inhibition rate of sevoflurane(1%,2%,5%)on hepatoma cells;si-con group(transfected si-con),si-CTRP6(transfected si-CTRP6),sevo+si-NC(transfected si-con,treated with 2%sevo),sevo+si-CTRP6 group(transfected si-CTRP6,treated with 2%sevo),transfected to HepG2 by liposome method;the expression of CTRP6 was detected by qRT-PCR.The migration and invasion of cells were detected by Transwell method.The protein expressions of CTRP6,Shh,Ihh and Smo were detected by Western blot.Results Compared with the control group,2%sevo and 5%sevo could significantly enhance the inhibition rate of liver cancer cells,and 2%sevo could significantly inhibit the migration and invasion of liver cancer cells(P<0.05);importantly,2%Sevo can down-regulate the expression of CTRP6 in hepatocarcinoma cells,and knockdown of CTRP6 can significantly inhibit the migration and invasion of hepatoma cells.Knockdown of CTRP6 can significantly inhibit the expression of key genes Shh,Ihh and Smo in Hedgehog signaling pathway.Conclusion Sevoflurane combined with down-regulation of CTRP6 can inhibit the migration and invasion of hepatoma cells.The mechanism is related to the inactivation of Hedgehog signaling pathway,which will provide experimental support for the treatment of hepatic carcinoma with sevoflurane and down-regulation of CTRP6.