摘要
背景与目的磷脂酰肌醇蛋白聚糖-5(glypican-5,GPC5)是一个重要的抑癌基因,然而GPC5对肺腺癌细胞增殖能力和基因表达的影响目前研究甚少。本研究拟在肺腺癌A549细胞中过表达GPC5以研究细胞增殖能力和基因表达变化情况。方法通过慢病毒载体构建稳定过表达GPC5的A549细胞株,通过Cell Counter Kit 8(CCK8)、平板克隆和Ed U实验检测细胞增殖能力;通过高通量转录组测序研究基因表达变化。结果相对于空白载体组,CCK8实验发现过表达GPC5可以明显抑制A549细胞的增殖速率;平板克隆实验结果显示,过表达GPC5之后A549细胞克隆形成能力下降(181±17 vs 278±23);Ed U染色结果显示过表达GPC5后阳性染色细胞比例下降。转录组测序结果提示过表达GPC5之后,2,108个基因表达发生明显变化,其中具有正性调节细胞增殖作用的基因明显下调。结论过表达GPC5可以明显抑制肺腺癌细A549的增殖能力,而且过表达GPC5后具有正性调节细胞增殖作用的基因表达下调。
Background and objective Glypican-5（GPC5） is an important tumor suppressor, while little is known about the impact of GPC5 on proliferation ability and gene expression in lung adenocarcinoma cell lines. Here, we stably overexpressed GPC5 in A549 cells and investigated the impact of cell proliferation ability and gene expression. Methods A549 cells that stably overexpressed GPC5 were constructed by lentivirus. Cell counter kit 8（CCK8）, colony formation, Ed U assay were conducted to analyze cell proliferation ability, and transcriptome sequencing was utilized to investigate gene expression profile. Results CCK8 assay showed that compared with empty vector, overexpression of GPC5 significantly inhibited cell proliferation rate in A549 cells and the number of colony was also decreased（181±17 vs 278±23）. Ed U assay also confirmed the percentage of positive staining cells decreased after GPC5 overexpression. Transcriptome sequencing revealed that 2,108 genes were differentially expressed after GPC5 overexpression. Among these differentially expressed genes, 47 genes of the Gene Ontology item ＂positive regulation of cell proliferation＂ were downregulated. Conclusion Overexpression of GPC5 inhibited proliferation ability in lung adenocarcinoma A549 cells and genes with the function of ＂positive regulation of cell proliferation＂ were downregulated.
出处
《中国肺癌杂志》
CAS
CSCD
北大核心
2016年第8期545-549,共5页
Chinese Journal of Lung Cancer
基金
国家自然科学基金项目(No.81372321和No.81572261)
江苏省重大疾病生物资源样本库项目(No.BM2015004)资助~~
