基于生信分析探索STARD8表达对肺癌A549细胞生物学功能的影响

Exploring the Impact of STARD8 Expression on the Biological Function of Lung Cancer A549 Cells Based on Bioinformatics Analysis

目的探讨STARD8对肺癌A549细胞增殖、迁移、侵袭和凋亡生物学功能的影响。方法从美国国家生物技术信息中心(NCBI),获取两份公开的基因芯片数据集(GSE18842和GSE68571),运用limma R包识别差异表达基因(DEGs)。采用最小绝对收缩和选择算子(LASSO)回归法结合SVM-RFE算法对候选基因进行筛选,构建受试者工作特征(ROC)曲线,应用CIBERSORT工具进行免疫浸润分析。通过RT-qPCR和Western blot分析STARD8在对照组及肺癌组的mRNA及蛋白水平,通过细胞平板克隆、Transwell、细胞划痕及流式细胞实验分析STARD8对A549细胞的增殖、侵袭、迁移及凋亡能力的影响。结果共筛选出285个上调基因和377个下调基因,取交集得到1个诊断标志物基因STARD8,免疫分析显示CD4 T细胞、NK细胞出现免疫抑制。肺癌组中STARD8的mRNA及蛋白表达均下调(P均<0.05)。STARD8过表达时肺癌A549细胞的增殖、侵袭及迁移能力降低,凋亡能力增加(P均<0.05)。结论STARD8可抑制肺癌A549细胞增殖、侵袭及迁移,促进细胞凋亡。

Objective To investigate the effects of STARD8 on the biological functions of proliferation,migration,invasion,and apoptosis in lung cancer A549 cells.Methods Two publicly available gene-on-a-chip datasets(GSE18842 and GSE68571)were acquired from the National Center for Biotechnology Information(NCBI).The limma R package was utilized to identify differentially expressed genes(DEGs).A combination of the minimum absolute contraction and selection operator(LASSO)regression method with the SVM-RFE algorithm was employed to screen candidate genes,receiver operating characteristic(ROC)curves was constructed,and immunoinfiltration analysis was performed by using the CIBERSORT tool.The mRNA and protein levels of STARD8 in both the control group and lung cancer group were assessed through RT-qPCR and Western blot.The impact of STARD8 on proliferation,invasion,migration,and apoptosis of A549 cells was evaluated via cell plate cloning,Transwell assay,scratch assay,and flow cytometry.Results A total of 285 up-regulated genes and 377 down-regulated genes were screened,and a diagnostic marker gene STARD8 was obtained by intersection.Immunoassay showed immunosuppression in CD4 T cells and NK cells.The mRNA and protein expression of STARD8 in the lung cancer group was down-regulated(P all<0.05).When STARD8 was overexpressed,the proliferation,invasion and migration ability of A549 cells of lung cancer decreased,while the apoptosis ability increased(P all<0.05).Conclusion STARD8 can inhibit the proliferation,invasion and migration of lung cancer A549 cells,and promote apoptosis.