摘要
目的观察长链非编码RNA HOTTIP对肺腺癌A549细胞侵袭转移能力的影响,通过上皮间充质转化(EMT)相关蛋白变化探讨其机制。方法将A549细胞分为HOTTIP-shRNA组、HOTTIP组和空载对照组,分别感染携带HOTTIP-shRNA的慢病毒、HOTTIP全长序列的慢病毒和空白慢病毒;采用Transwell实验观察细胞侵袭能力,细胞划痕实验观察细胞迁移能力,Western blotting法检测细胞间质标志蛋白Vimentin、上皮标志蛋白E-cadherin。结果HOTTIP-shRNA组、HOTTIP组、空载对照组穿膜细胞的数量分别为(12.85±3.25)、(305.65±53.45)、(45.25±10.75)个,细胞迁移率为35.62%±10.45%、13.84%±4.12%、64.52%±14.24%,HOTTIP-shRNA组穿膜细胞数量、细胞迁移率低于空载对照组,而HOTTIP组穿膜细胞数量、细胞迁移率均高于HOTTIP-shRNA组和空载对照组(P均<0.05)。与空载对照组比较,HOTTIP-shRNA组细胞Vimentin表达减少、E-cadherin表达增加(P均<0.05);与HOTTIP-shRNA组和空载对照组比较,HOTTIP组细胞Vimentin表达增加、E-cadherin表达减少(P均<0.05)。结论HOTTIP可增强肺腺癌A549细胞侵袭转移能力,与其调控E-cadherin、Vimentin蛋白表达促进EMT形成有关。
Objective To observe the effect of long non-coding RNA HOTTIP on the invasion and metastasis of lung adenocarcinoma A549 cells,and to explore the mechanism of the change of epithelial-mesenchymal transition(EMT)-related proteins.Methods A549 cells were divided into the HOTTIP-shRNA group,HOTTIP group,and empty control group,respectively,which were infected with HOTTIP-shRNA-bearing lentivirus,HOTTIP full-length lentivirus,and blank lentivirus;Transwell assay was used to observe cell invasion ability,and the cell migration ability was observed by scratch test.The cell interstitial marker protein Vimentin and epithelial marker protein E-cadherin were detected by Western blotting.Results The number of transmembrane cells in the HOTTIP-shRNA group,HOTTIP group and empty control group was 12.85±3.25,305.65±53.45,and 45.25±10.75,and the cell migration rates were 35.62%±10.45%,13.84%±4.12%,and 64.52%±14.24%;the number of transmembrane cells and cell migration rate in HOTTIP-shRNA group were lower than those in the empty control group,while the number of transmembrane cells and cell migration rate were higher in the HOTTIP group than in the HOTTIP-shRNA group and empty control group(both P<0.05).Compared with the empty control group,the expression of Vimentin and the expression of E-cadherin increased in the HOTTIP-shRNA group(both P<0.05).Compared with the HOTTIP-shRNA group and empty control group,Vimentin expression increased and E-cadherin expression decreased in the HOTTIP group(both P<0.05).Conclusion HOTTIP can enhance the invasion and metastasis of lung adenocarcinoma A549 cells,which is related to the regulation of E-cadherin and Vimentin protein expression and the promotion of EMT formation.
作者
金文静
杨华
王林宣
李珊珊
顾文超
JIN Wenjing;YANG Hua;WANG Linxuan;LI Shanshan;GU Wenchao(The People′s Hospital of Pudong New Area,Shanghai University of Health and Science,Shanghai 201200,China)
出处
《山东医药》
CAS
2020年第2期18-21,共4页
Shandong Medical Journal
基金
上海市浦东新区卫生系统重点学科建设项目(PWZxk2017-24)
