Sestrin2在miR-19诱导的非小细胞肺癌细胞上皮-间质转化中的表达及六神丸干预研究

目的 探讨Sestrin2在六神丸调控miR-19诱导的非小细胞肺癌细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)中的作用.方法 实验分为对照组、miR-19过表达组、六神丸组.对照组为常规血清培养的A549细胞;miR-19过表达组为常规血清培养的过表达miR-19 A549细胞;六神丸组予以六神丸含药血清干预过表达miR-19 A549细胞.48 h后分别以CCK-8检测A549细胞增殖,Transwell小室检测A549细胞迁移能力,实时荧光定量PCR(quantitative Real-time PCR,qRT-PCR)检测miR-19表达,蛋白印迹检测Sestrin2、PTEN以及EMT标记分子E-cadherin、Vimentin的表达.结果 细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测结果显示,六神丸组A549细胞增殖速度较对照组、miR-19过表达组明显减慢;Transwell小室实验显示,六神丸组A549细胞迁移数量较miR-19过表达组、对照组明显减少;qRT-PCR检测结果显示,六神丸组A549细胞中miR-19表达较miR 19过表达组、对照组明显降低;Western blot检测结果显示,与对照组、miR-19过表达组相比,六神丸组A549细胞中Sestrin2、E-cadherin、PTEN表达明显上调,Vimentin表达明显下调.结论 六神丸能够抑制miR-19诱发的人肺腺癌细胞株A549发生EMT,与激活Sestrin2信号通路有关.

Study on expression of Sestrin2 in epithelial-mesenchymal transition of non-small cell lung cancer cells induced by miR-19 and intervention of Liushen pills

Objective To investigate the role of Sestrin2 in the regulation of miR-19-induced epithelial-mesenchymal transition (EMT) in non-small cell lung cancer cells.Methods The experiment had a control group,an miR-1 9 overexpression group,and a Liushen pills group.The control group was normal serumcultured A549 cells,the miR-19 overexpression group was normal serum-cultured miR-19 overexpressing A549 cells,and the Liushen pill group was Liushen pills-containing serum-cultured the miR-19 overexpression of A549 cells.CCK-8 was used to detect A549 cell proliferation,Transwell chamber was used to detect A549 cell migration capability,real-time quantitative PCR (RT-PCR) was used to detect the miR-19 expression,and western blot was used to detect the protein levels of Sestrin2,PTEN,and EMT markers including E-cadherin and Vimentin in 48 hours later.Results The cell counting kit-8 (CCK-8) showed that the proliferation rate ofA549 cells was significantly slower in the Liushen pill group than in the control group and the miR-19 overexpression group.Transwell chamber test showed that the number of A549 cell migration was significantly lower in the Liushen pill group than in the miR-19 overexpression group and the control group.qRT-PCR showed that the miR-19 expression of A549 cells was significantly lower in the Liushen pill group than in the miR-19 overexpression group and the control group.Western blot showed that the expressions of Sestrin2,E-cadherin,and PTEN was significantly more up-regulated and that of Vimentin was more down-regulated in A549 cells in the Liushen pill group than in the control group and the miR-19 overexpression group.Conclusion Liushen pills can inhibit EMT in human lung adenocarcinoma cell line A549 induced by miR-19,which is related to the activation of Sestrin2 signaling pathway.