天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2015年
8期
856-859
,共4页
胃肿瘤%腺癌%细胞增殖%细胞周期%RNA干扰%MACC1基因%MGC-803细胞
胃腫瘤%腺癌%細胞增殖%細胞週期%RNA榦擾%MACC1基因%MGC-803細胞
위종류%선암%세포증식%세포주기%RNA간우%MACC1기인%MGC-803세포
stomach neoplasms%adenocarcinoma%cell proliferation%cell cycle%RNA interference%MACC1%MGC-803 cell
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。
目的：研究下調MACC1錶達對胃腺癌細胞株MGC-803生長作用的影響。方法設計併閤成RNAi榦擾片段，脂質體介導MACC1-siRNA1（MACC1-siRNA1組）和MACC1-siRNA2（MACC1-siRNA2組）轉染MGC-803細胞，同時設非特異性榦擾片段為對照組。應用qRT-PCR檢測轉染後各組MACC1的mRNA錶達，噻唑藍比色實驗、平闆剋隆形成實驗和流式細胞週期實驗檢測RNAi轉染後MGC-803細胞增殖能力的變化，Western blot檢測轉染後MACC1、P21、CDK4、CCND1和c-myc蛋白的錶達，併進行比較分析。結果與對照組相比，MACC1-siRNA1組和MACC1-siRNA2組MACC1錶達在mRNA及蛋白水平下降，細胞的增殖速度變慢，單剋隆形成數量減少，細胞週期中G1期細胞的比例顯著升高，P21的錶達增加，MACC1、CDK4、CCND1和c-myc的錶達減少。結論下調MACC1能使MGC-803細胞的細胞週期進程受阻，抑製細胞的增殖，MACC1可以作為治療胃癌的有效靶點。
목적：연구하조MACC1표체대위선암세포주MGC-803생장작용적영향。방법설계병합성RNAi간우편단，지질체개도MACC1-siRNA1（MACC1-siRNA1조）화MACC1-siRNA2（MACC1-siRNA2조）전염MGC-803세포，동시설비특이성간우편단위대조조。응용qRT-PCR검측전염후각조MACC1적mRNA표체，새서람비색실험、평판극륭형성실험화류식세포주기실험검측RNAi전염후MGC-803세포증식능력적변화，Western blot검측전염후MACC1、P21、CDK4、CCND1화c-myc단백적표체，병진행비교분석。결과여대조조상비，MACC1-siRNA1조화MACC1-siRNA2조MACC1표체재mRNA급단백수평하강，세포적증식속도변만，단극륭형성수량감소，세포주기중G1기세포적비례현저승고，P21적표체증가，MACC1、CDK4、CCND1화c-myc적표체감소。결론하조MACC1능사MGC-803세포적세포주기진정수조，억제세포적증식，MACC1가이작위치료위암적유효파점。
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.
