摘要
目的分析Rab32对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖、迁移及侵袭的影响及作用机制;方法通过生物信息学判断NSCLC中Rab32的表达;用qRT-PCR和Western blot检测NSCLC细胞中Rab32 mRNA和蛋白表达水平;应用Rab32过表达慢病毒感染A549和H1299细胞,构建Rab32稳转A549和H1299细胞株为观察组,空载体慢病毒感染A549和H1299细胞后构建的为对照组;利用CCK-8和集落形成实验判断Rab32对NSCLC细胞影响增殖能力;通过划痕和Transwell实验检测Rab32对A549和H1299细胞迁移和侵袭能力的影响;利用Western blot检测不同组细胞中EMT标志物(E-cadherin、N-cadherin和Vimentin)蛋白的表达水平变化。结果通过生物信息学分析显示,肺腺癌(Lung adenocarcinoma,LUAD)和肺鳞癌(lung squamous cell carcinoma,LUSC)中Rab32的mRNA表达较对照组显著降低(P<0.05);qRT-PCR及Western blot结果表明A549、H1299和HCC827中Rab32 mRNA和蛋白表达水平显著低于正常人支气管上皮细胞Beas-2B,其中A549和H1299细胞中Rab32 mRNA和蛋白较Beas-2B下降显著。A549、H1299细胞中,观察组的Rab32 mRNA和蛋白表达水平显著高于空载体对照组(P<0.05)。CCK-8和集落形成实验结果表明观察组的细胞增殖和克隆能力较对照组显著下降(P<0.05);划痕和Transwell实验结果提示观察组细胞的迁移和侵袭能力明显下降(P<0.05);Western blot结果显示观察组细胞的上皮表型标志物E-cadherin蛋白表达水平显著高于对照组,而间皮表型标志物N-cadherin和Vimentin表达较对照组明显升高(P<0.05)。结论Rab32抑制NSCLC细胞的增殖、迁移和侵袭,可作为潜在临床治疗NSCLC患者新途径。
Objective To investigate the role and novel mechanism of Rab32 on the proliferation,migration,and invasion of non-small cell lung cancer(NSCLC)cells.Methods The expression of Rab32 in NSCLC was assessed using bioinformatics.The mRNA and protein expression levels of Rab32 in NSCLC cells were determined by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot analysis.Stable Rab32-overexpressing A549 and H1299 cell lines were constructed using a lentivirus system,with the overexpression group serving as the observation group.The effects of Rab32 on the proliferation of NSCLC cells were evaluated by CCK-8 and colony formation assays.The impact of Rab32 on the migration and invasion capabilities of A549 and H1299 cells was examined using scratch and Transwell assays.The expression levels of epithelial-mesenchymal transition(EMT)markers(E-cadherin,N-cadherin,and Vimentin)in different groups of cells were measured by Western blot.Results Bioinformatics analysis indicated that the mRNA expression of Rab32 was significantly reduced in lung adenocarcinoma(LUAD)and lung squamous cell carcinoma(LUSC)compared to the control group(P<0.05).qRT-PCR and Western blot results revealed that the mRNA and protein expression levels of Rab32 in A549,H1299,and HCC827 were significantly lower than those in normal human bronchial epithelial cells(Beas-2B),with more pronounced decreases in A549 and H1299 cells.In A549 and H1299 cells,the mRNA and protein expression levels of Rab32 in the observation group were significantly higher than those in the empty vector control group(P<0.05).The CCK-8 and colony formation assay results showed that the cell proliferation and cloning capacity of the observation group were significantly reduced compared to the control group(P<0.05).The scratch and Transwell assay results suggested a marked decrease in the migration and invasion capabilities of the observation group cells(P<0.05).Western blot analysis showed that the protein expression level of the epithelial phenotype marker E-cadh
作者
赵蒙蒙
黄洁
余荣环
王葆青
Zhao Mengmeng;Huang Jie;Yu Ronghuan;Wang Baoqing(Department of Respiratory Medicine,Shanghai Xuhui Central Hospital,Zhongshan-Xuhui Hospital,Fudan University,Shanghai 200031,China;Department Respiratory Medicine,Zhongshan Hospital,Fudan University,Shanghai 200032,China)
出处
《中华肺部疾病杂志（电子版）》
2024年第4期512-518,共7页
Chinese Journal of Lung Diseases(Electronic Edition)
基金
上海市徐汇区卫生系统高峰学科建设项目(SHXHZDXK202312)。
关键词
非小细胞肺癌
Rab32
细胞增殖
细胞迁移
侵袭能力
Non-small cell lung cancer
Rab32
Cell proliferation
Cell migration
Invasive capacity
