天津医药 ›› 2017, Vol. 45 ›› Issue (7): 677-681.doi: 10.11958/20170352

• 细胞与分子生物学 • 上一篇    下一篇

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

陈翠翠,梁焕坤,李康燕,程承武,刘细潘 ,黎杰星,钟树海,张立成,李来庆   

  1. 1 广州优迪生物科技股份有限公司 (邮编 510663); 2 解放军第 88 医院肿瘤科
  • 收稿日期:2017-03-20 修回日期:2017-05-04 出版日期:2017-07-15 发布日期:2017-08-08
  • 通讯作者: 李来庆 E-mail:121642202@qq.com
  • 基金资助:
    广东省科技计划项目 (2016A010119067); 山东省医药卫生科技发展计划项目 (2016WS0725)

Effects of overexpression of miR-30b on the biological function and tumor formation of human gastric cancer cells

CHEN Cui-cui,LIANG Huan-kun,LI Kang-yan,CHENG Cheng-wu,LIU Xi-pan,LI Jie-xing,ZHONG Shu-hai,ZHANG Li-cheng,LI Lai-qing   

  1. 1 Guangzhou Youdi Biotechnology Co., Ltd., Guangzhou 510663, China;2 Department of Oncology, Eighty-eighth Hospital of PLA
  • Received:2017-03-20 Revised:2017-05-04 Published:2017-07-15 Online:2017-08-08

摘要: 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。

关键词: 微 RNAs, 细胞增殖, 细胞周期, 细胞凋亡, 肿瘤侵润, miR-30b

Abstract: 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 eIF5A2 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 miRcontrol were injected into nude mice to observe the tumor formation and the expression of eIF5A2 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.

Key words: microRNAs, cell proliferation, cell cycle, apoptosis, neoplasm invasiveness, miR-30b

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