miR-199-3p过表达调控肺癌A549细胞生长、侵袭及上皮间质转化机制研究  

Mechanism of miR-199-3p Overexpression in Regulating Growth,Invasion and Epithelial-mesenchymal Transformation of Lung Cancer A549 Cells

作　　者：熊伟杰[1] 王云涛[1] 何朗 XIONG Wei-jie;WANG Yun-tao;HE Lang(Department of Oncology,the Fifth People's Hospital of Chengdu,Chengdu 611130,China)

机构地区：[1]成都市第五人民医院肿瘤科,成都611130

出　　处：《解放军医药杂志》2022年第4期13-18,共6页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：四川省卫生和计划生育科研课题项目(18PJ035)。

摘　　要：目的探究miR-199-3p过表达靶向抑制SOX5对肺癌A549细胞生长、侵袭及上皮间质转化(EMT)的影响。方法荧光素酶实验检测miR-199a-3p与SOX5靶向关系。构建SOX5 pcDNA载体过表达SOX5,将miR-199-3p与pcDNA-SOX5单独或联合转染A549,将细胞随机分为对照组(Control组)、miR-199-3p过表达组(mimic组)、SOX5组和mimic+SOX5组进行后续实验。克隆形成法检测肺癌A549细胞增殖情况,流式细胞术检测肺癌A549细胞凋亡情况,Transwell检测肺癌A549细胞侵袭能力;荧光显微镜观察肺癌A549细胞EMT形态学变化;蛋白免疫印迹试验检测Ki-67、PCNA、Bax、Bcl-2、Caspase-3、E-cadherin、N-cadherin、Vimentin蛋白表达水平。结果与SOX5组比较,mimic+SOX5组细胞克隆形成率降低,Ki-67、PCNA蛋白水平降低,细胞凋亡率升高,Bax/Bcl-2和cleaved Caspase-3/Caspase-3比值升高,细胞侵袭数降低,E-cadherin蛋白水平升高,N-cadherin、Vimentin蛋白水平降低(P<0.05)。结论miR-199-3p靶向SOX5抑制肺癌A549细胞增殖、侵袭和EMT,并诱导细胞凋亡。Objective To investigate effects of miR-199-3p overexpression in targeting and inhibiting SOX5 on growth,invasion and epithelial mesenchymal transformation(EMT)of lung cancer A549 cells.Methods Luciferase assay was used to detect the targeting relationship between miR-199a-3p with SOX5.SOX5 was overexpressed by the SOX5 pcDNA vector,and A549 was transfected with miR-199-3p and pcdna-SOX5 alone or in combination.The cells were randomly divided into Control group,miR-199-3p overexpression group(mimic group),SOX5 group and mimic+SOX5 group for subsequent experiments.Proliferation conditions of lung cancer A549 cells were detected by clone formation method,and cell apoptosis conditions were detected by flow cytometry.Transwell experiment was used to detect invasion ability of A549 cells,and morphological changes of EMT were observed by fluorescence microscopy.Expressions of Ki-67,PCNA,Bax,Bcl-2,Caspase-3,E-cadherin,N-cadherin and Vimentin proteins were detected by Western blot method.Results Compared with those in SOX5 group,clone formation rate of cells as well as protein levels of Ki-67 and PCNA were significantly decreased,while apoptosis rate and ratios of Bax/Bcl-2 and cleaved Caspase-3/Caspase-3 were significantly increased;the number of invasive cells was significantly decreased;level of E-cadherin protein was significantly increased,while levels of N-cadherin and Vimentin proteins were significantly increased in mimic+SOX5 group(P<0.05).Conclusion The miR-199-3p targeted SOX5 may inhibit proliferation,invasion and EMT,and it may also induce cell apoptosis of lung cancer A549 cells.

关 键 词：肺肿瘤 A549细胞 细胞增殖 细胞凋亡 钙黏着糖蛋白类 SOXD转录因子类 上皮间质转化 

分 类 号：R734.2[医药卫生—肿瘤]