天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (8): 856-859.doi: 10.11958/j.issn.0253-9896.2015.08.007

• 细胞与分子生物学 • 上一篇    下一篇

MACC1基因下调对胃腺癌细胞的生长抑制作用

罗毅，刘路培，于路   

  1. 广西科技大学附属柳州市人民医院急症科
  • 收稿日期:2014-06-18 修回日期:2014-12-09 出版日期:2015-08-15 发布日期:2015-08-15
  • 作者简介:罗毅（1978），男，副主任医师，硕士，主要从事胃肠道肿瘤发病机制研究

Effect of MACC1 down-regulation on proliferation of gastric adenocarcinoma cell line

LUO Yi, LIU Lupei, YU Lu   

  1. Department of Emergency, Liuzhou People′s Hospital Affiliated to Guangxi University of Technology, Guangxi 545006, China
  • Received:2014-06-18 Revised:2014-12-09 Published:2015-08-15 Online:2015-08-15

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摘要： 目的研究下调MACC1 表达对胃腺癌细胞株MGC-803 生长作用的影响。方法设计并合成RNAi 干扰片段，脂质体介导MACC1-siRNA1（MACC1-siRNA1 组）和MACC1-siRNA2（MACC1-siRNA2 组）转染MGC-803 细胞，同时设非特异性干扰片段为对照组。应用qRT-PCR 检测转染后各组MACC1 的mRNA 表达，噻唑蓝比色实验、平板克隆形成实验和流式细胞周期实验检测RNAi 转染后MGC-803 细胞增殖能力的变化，Western blot 检测转染后MACC1、P21、CDK4、CCND1 和c-myc 蛋白的表达，并进行比较分析。结果与对照组相比，MACC1-siRNA1 组和MACC1-siRNA2 组MACC1 表达在mRNA 及蛋白水平下降，细胞的增殖速度变慢，单克隆形成数量减少，细胞周期中G1 期细胞的比例显著升高，P21 的表达增加，MACC1、CDK4、CCND1 和c-myc 的表达减少。结论下调 MACC1 能使MGC-803 细胞的细胞周期进程受阻，抑制细胞的增殖，MACC1 可以作为治疗胃癌的有效靶点。

关键词: 胃肿瘤, 腺癌, 细胞增殖, 细胞周期, RNA 干扰, MACC1 基因, MGC-803 细胞

Abstract: Objective To study the effects of MACC1 down- regulation on the growth of gastric adenocarcinoma cells. Methods siRNA (MACC1-siRNA1 and MACC1-siRNA2) that can transiently silenced MACC1 was designed, syn⁃ thesized and transfected into MGC-803 cells by lipofectamine 2000. Non-specific siRNA was transfected to be used as nega⁃ tive control. The efficiency of MACC1 depletion was determined by Real-time quantitative PCR. MTT, colony formation and flow cytometry assay were performed to examine cell proliferation. The expressions of MACC1, P21,CDK4, CCND1 and cmyc were determined by Western blot. Results Compared with cells in negative control group, transiently silencing MACC1 decreased the expression of MACC1 in MGC-803 cells shown by Real-time PCR. MACC1 downregulation drastical⁃ ly changed the proliferation, colony formation and cell cycle of gastric adenocarcinoma cells in vitro (P＜0.05). The expres⁃ sions of MACC1 , CDK4, CCND1 and c-myc proteins in cells of MACC1 silence group were much lower while P21 expres⁃ sion level was much higher than those in negative control. Conclusion Down-regulation of MACC1 result in blocking cell cycle, inhibiting proliferation of MGC-803 cells. So it may serve as a promising target in the treatment of gastric cancer.

Key words: stomach neoplasms, adenocarcinoma, cell proliferation, cell cycle, RNA interference, MACC1, MGC-803 cell