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  

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

作　　者：鄢文[1] 靳文[1] 王昂[1] 宋维舒[1] Yan Wen Jin Wen WangAng Song Weishu(First Department of Oncology, Guangdong No.2 Provincial People＇s Hospital, Guangzhou 510317, China)

机构地区：[1]广东省第二人民医院肿瘤一科,广州510317

出　　处：《国际医药卫生导报》2017年第5期626-629,636,共5页International Medicine and Health Guidance News

基　　金：国家自然科学基金资助项目（81302965）

摘　　要：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一间质转化（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-19A549细胞；六神丸组予以六神九含药血清干预过表达miR-19A549细胞。48h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Sestrin 2、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。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-19 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 ceils 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.

关 键 词：六神丸 上皮一间质转化 Sestrin2 miR-19 

分 类 号：R734.2[医药卫生—肿瘤]