9-顺式维甲酸诱导胃癌MGC803细胞周期阻滞和凋亡的作用机制  

Inducement Effect of 9-Cis Retinoic Acid on Cell Cycle Arrest and Apoptosis of Gastric Carcinoma Cell Line MGC803 and Its Mechanism

作　　者：徐瑞成[1] 周艳红[1] 呼文亮[1] 

机构地区：[1]中国人民武装警察部队医学院细胞生物学教研室,天津300162

出　　处：《癌症》2006年第12期1483-1487,共5页Chinese Journal of Cancer

基　　金：天津市自然科学基金项目(No.013804311)~~

摘　　要：背景与目的:9-顺式维甲酸(9-cisretinoicacid,9-cisRA)对胃癌的抗癌活性及机制尚不清楚,本研究以MGC803为靶细胞观察9-cisRA对胃癌的作用。方法:采用RT-PCR法检测维甲类X受体α(recinoidXreceptor!,RXRα)、细胞周期蛋白CyclinD1和细胞周期蛋白依赖性激酶CDK4mRNA表达,流式细胞术检测细胞周期,MTT实验检测9-cisRA作用MGC803细胞后生长抑制情况,Hoechst33342/PI双荧光染色和琼脂糖凝胶电泳检测凋亡,免疫细胞化学检测凋亡相关基因Bcl-2蛋白表达。结果:0.1～10μmol/L9-cisRA作用MGC803细胞96h,能显著抑制细胞增殖。10μmol/L9-cisRA分别作用48、72和96h,G1期细胞随着作用时间的延长而增加,呈明显的G1期阻滞。作用72h后,细胞出现核染色质凝集、DNA片段化等凋亡特征;从作用48h开始,Bcl-2表达即显著下降。在MGC803细胞中RXRα表达较弱,经10μmol/L9-cisRA作用48h其表达水平显著增加(P<0.01);作用96h,细胞中CyclinD1和CDK4表达显著降低(P<0.01)。结论:9-cisRA能明显诱导MGC803细胞周期G1期阻滞和凋亡,该作用可能与其下调细胞周期因子CyclinD1和CDK4表达有关。BACKGROUND ＆ OBJECTIVE：Antitumor effect of 9-cis retinoic acid （9-cis RA） on gastric carcinoma is unclear yet. This study was to explore the inducement effect of 9-cis RA on cell cycle arrest and apoptosis of gastric carcinoma cell line MGC803 and its mechanism. METHODS： The expression of RXRa, Cyclin D1, and CDK4 in MGC803 cells was detected by reverse transcription-polymerase chain reaction （RT-PCR）. Cell cycle was detected by flow cytometry. The growth inhibition was analyzed by MTT assay. The apoptosis was detected by agarose gel electrophoresis and Hoechst33342/PI staining. The expression of apoptosis-associated gene Bcl-2 was detected by SP immunocytochemistry. RESULTS. When treated with 0.1-10 μmol/L 9-cis RA for 96 h, the proliferation of MGC803 cells was significantly inhibited. The proportion of MGC803 cells at G1 phase was significantly increased when treated with 10 μmol/L 9-cis RA for 48, 72, and 96 h, and showed an apparent G1 phase arrest. When treated with 9-cis RA for 72 h, typical apoptotic changes, such as chromatin condensation and DNA ladder, were observed in MGC803 cells. The expression of Bcl-2 was significantly decreased in MGC803 cells when treated with 10 μmol/L 9-cis RA for 48 h. RXRa expression was at a low level in MGC803 cells and upregulated when treated with 10 μmol/L 9-cis RA for 48 h （P〈0.01）. The expression of Cyclin D1 and CDK4 in MGC803 cells was both significantly down-regulated when treated with 10 μmol/L 9-cis RA for 96 h （P〈0.01）. CONCLUSION： 9-Cis RA could induce G1 phase arrest and apoptosis in MGC803 cells through down-regulating the expression of cell cycle factors Cyclin D1 and CDK4.

关 键 词：胃肿瘤 MGC803细胞 9-顺式维甲酸 细胞周期 RXRα 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]