摘要
目的通过下调mir-151a-5p在肺癌A549细胞中的表达,研究其对细胞增殖、迁移、侵袭能力和化疗敏感性的影响。方法应用RT-qPCR检测mir-151a-5P在肺腺癌组织和肺腺癌细胞株以及转染inhibitor后的表达情况。通过CCK-8、Transwell、克隆形成实验分别检测转染inhibitor后A549细胞的增殖、迁移、侵袭能力以及克隆形成能力的变化。Western blot检测转染后EMT相关蛋白的表达。CCK-8、流式细胞术检测转染后A549细胞对顺铂的敏感性变化。结果 mir-151a-5p在人肺腺癌组织中的表达较正常肺组织明显升高(P<0.01),且在4种肺癌细胞株(H1975、H1299、H2228、A549)中,A549细胞中mir-151a-5p表达量较高(P<0.01)。成功构建了mir-151a-5p-inhibitor转染的A549细胞(P<0.01)。mir-151a-5p inhibitor转染组较NC对照组细胞的增殖能力、迁移侵袭能力以及克隆形成能力都明显降低(P<0.05)。Western blot检测结果显示,与NC对照组比较,mir-151a-5p inhibitor转染组E-cadherin表达明显升高,而N-cadherin、Vimentin、Notch1表达明显下降(P<0.01)。CCK-8结果显示mir-151a-5p inhibitor转染组IC_(50)显著低于NC对照组(P<0.01)。流式细胞术检测结果显示,加入顺铂后mir-151a-5p inhibitor转染组凋亡率显著高于NC对照组(P<0.01)。结论下调mir-151a-5p能显著抑制A549细胞的增殖、迁移及侵袭能力,并能够增加对顺铂的化疗敏感性。
Objective To investigate the effects of mir-151 a-5 p down-regulation on the proliferation,migration,invasion and chemosensitivity of A549 cells.Methods RT-qPCR was used to detect the expression of mir-151 a-5 P in 16 surgical specimens of lung adenocarcinoma and in different lung adenocarcinoma cell lines(H1975,H1299,H2228,A549 and HBE).CCK-8 assay,Transwell test and clonogenic assay were used to assess the changes in the proliferation,migration,invasion and clonogenic ability of A549 cells after transfection with a mir-151 a-5 p inhibitor.Western blotting was performed to detect the expression of epithelial-mesenchymal transition(EMT)-related proteins in the transfected cells;CCK-8 assay and flow cytometry were used to evaluate the changes in cisplatin sensitivity of A549 cells after the transfection.Results The expression of mir-151 a-5 p was significantly increased in human lung adenocarcinoma tissues as compared with normal lung tissues(P<0.01).Among the 4 lung adenocarcinoma cell lines,A549 cells expressed the highest levels of mir-151 a-5 p(P<0.01).Transfection with the mir-151 a-5 p inhibitor,compared with the negative control,significantly attenuated the proliferation,migration,invasion and clonogenic ability of A549 cells(P<0.05).Western blotting revealed a significant enhancement of E-cadherin expression and obviously lowered expressions of N-candherin,vimentin and Notch1 in A549 cells transfected with the mir-151 a-5 p inhibitor(P<0.01).CCK-8 assay showed that the IC50 of cisplatin was significantly lowered in mir-151 a-5 p inhibitor-transfected cells as compared with the negative control group(P<0.01);the transfection also resulted in a significantly increased apoptotic rate of the cells in response to cisplatin treatment(P<0.01).Conclusion Down-regulation of mir-151 a-5 p expression can significantly inhibit the proliferation,migration and invasion of A549 cells and increase their chemosensitivity to cisplatin.
作者
隆姝孜
周寒静
刘静姝
刘洪良
万林
周丽
张涛
LONG Shuzi;ZHOU Hanjing;LIU Jingshu;LIU Hongliang;WAN Lin;ZHOU Li;ZHANG Tao(Department of Oncology,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China;Department of Gastrointestinal Surgery,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2019年第8期770-777,共8页
Journal of Third Military Medical University
基金
重庆市卫生计生委医学科研计划项目(2017MSXM005)~~
