过表达miR-30b对人胃癌细胞株SGC-7901和AGS生物学功能及其肿瘤形成的影响

摘要

Objective To investigate the effect of overexpression of miR-30b on the proliferation,cell cycle,apoptosis and invasion of gastric cancer cell line SGC-7901 and AGS,and the inhibitory effect on the tumor formation in vivo.Methods SGC-7901 and AGS cells were transfected with miR-30b mimics and miR-control,and qRT-PCR was used to detect the expression levels of miR-30b.Western blot assay was used to detect the expression of eIFSA2 protein.CCK-8 assay was used to measure the cell proliferation.Flow cytometry was used to analyze cell cycle and apoptosis.Transwell assay was used to detect cell invasion.In addition,the SGC-7901 and AGS cells transfected with miR-30b mimics and miR-control were injected into nude mice to observe the tumor formation and the expression of eIFSA2 protein in vivo.Results Results of qRT-PCR showed that the relative expression of miR-30b was significantly higher than that of miR-control group (P ＜ 0.05).Western blot assay showed that the expression of eIF5A2 protein was decreased in miR-30b mimics group.CCK-8 assay showed that cell proliferation was inhibited in miR-30b mimics group.The result of flow cytometry showed that the cell cycle decreased and the apoptosis increased in miR-30b group.Transwell assay showed that the cell invasion was significantly lower in miR-30b group than that of control group (P ＜ 0.05).Overexpression of miR-30b inhibited the formation of tumor and decreased the expression of eIF5A2 protein in vivo.Conclusion Overexpression of miR-30b inhibits the proliferation,invasion and tumor formation of gastric cancer cells,and reduces the expression of eIF5A2 protein,which provides a potential target for gastric cancer treatment.%目的 研究过表达miR-30b对人胃癌细胞株SGC-7901和AGS的增殖、细胞周期、凋亡、侵袭等生物学功能的影响及对体内肿瘤形成的抑制作用.方法 SGC-7901和AGS细胞中转染miR-30b类似物和miR-对照后,qRT-PCR检测转染后细胞miR-30b的表达变化;Western blot检测eIF5A2蛋白的表达;CCK-8检测SGC-7901和AGS细胞增殖;流式细胞术检测细胞周期和凋亡;Transwell实验检测细胞体外侵袭能力.转染miR-30b类似物和miR-对照的SGC-7901和AGS细胞,分别注射裸鼠体内,观察肿瘤的形成及肿瘤组织中eIF5A2蛋白的表达.结果 qRT-PCR结果表明miR-30b类似物组的相对表达量显著高于miR-对照组(P＜ 0.05);Western blot结果显示,miR-30b类似物组的eIF5A2蛋白表达降低;CCK-8实验表明miR-30b类似物组中细胞增殖受到抑制;流式细胞术显示miR-30b类似物组的细胞周期减慢,凋亡增加;在Transwell迁移实验中,miR-30b类似物组细胞侵袭能力明显低于miR-对照组(P＜0.05).细胞体内成瘤实验显示,miR-30b过表达抑制肿瘤的形成,肿瘤组织中eIF5A2蛋白表达降低.结论 miR-30b过表达抑制人胃癌细胞的增殖、侵袭和肿瘤的形成,降低eIF5A2蛋白的表达,为胃癌治疗提供了一个潜在的靶点.