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。%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 c-myc 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.
展开▼
