地西他滨对裸鼠子宫内膜癌移植瘤的作用及机制研究  

The Study of Decitabine Effect on the Endometrial Carcinoma Xenografted in Nude Mice

作　　者：李冉红[1] 王雪萍[2] 刘辉[1] 

机构地区：[1]四川大学华西第二医院妇产科出生缺陷与相关妇儿疾病教育部重点实验室,成都610041 [2]德阳市人民医院妇产科,德阳618000

出　　处：《四川大学学报（医学版）》2016年第6期843-847,共5页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技支撑计划项目(No.2010SZ0107)资助

摘　　要：目的探究去甲基化药物地西他滨对裸鼠子宫内膜癌移植瘤的作用及其机制。方法建立子宫内膜癌裸鼠移植瘤模型后随机分为地西他滨组(AZA组)、顺铂组(DDP组)、醋酸甲羟孕酮组(MPA组)、AZA+DDP组、AZA+MPA组、DDP+MPA组、模型组,每组3只,各实验组给予相应药物(1μg/g单用或各1μg/g联合),模型组给予生理盐水,每3d一次尾静脉注射给药,共给药8次。处理后观察各组裸鼠瘤体生长情况;取出瘤体后,计算抑瘤率;采用甲基化特异性PCR(MSP)、Western blot和TUNEL染色法分别检测移植瘤组织细胞中RASSF1A基因启动子区域甲基化状态、蛋白表达及肿瘤细胞的凋亡情况。结果 AZA+DDP组抑瘤率最高,而AZA组抑瘤率最低。AZA组、AZA+DDP组、AZA+MPA组RASSF1A基因启动子区域甲基化水平明显降低,显示明显的非甲基化条带,其余组则主要显示甲基化条带。AZA组、AZA+DDP组、AZA+MPA组这三组之间RASSF1A蛋白的表达量差异无统计学意义(P>0.05),但均高于模型组(P<0.05);DDP组、MPA组、DDP+MPA组与模型组比较差异无统计学意义(P>0.05)。凋亡指数为模型组<3个单药组<3个联合用药组(P<0.05)。结论地西他滨可逆转内膜癌中RASSF1A基因启动子区域的异常甲基化,恢复RASSF1A蛋白生物学功能,增强DDP与MPA的药效,对临床子宫内膜癌的治疗有很大的潜在应用价值。Objective To explore the effect of the demethylation drug 5-Aza-CdR on endometrial carcinoma xenografted in nude mice. Methods Randomly assigned the mice into decitabine （AZA）, cisplatin （DDP）, medroxyprogesterone acetate （MPA）, AZA＋DDP, AZA＋ MPA, DDP＋MPA and model groups （three in each group） after building the models of xenografted tumor by transplanting the HEC-1B cells on nude mice, and dealt them respectively with corresponding drugs （1 μg/g, single or combination） in the experiment groups and normal saline in model group （injected per 3 d, 8 injections in total）. Then the tumor inhibitory rates in different groups were calculated. The methylation and protein expression of RASSF1A gene was estimated by methylation specific PCR （MSP） and Western blot respectively, and apoptosis situation of carcinoma cell was estimated by tunel. Results Inhibitory rate in AZA ＋ DDP group was the highest, and the lowest was AZA group. RASSF1A gene promoter region methylation levels of AZA, AZA ＋DDP and AZA ＋ MPA groups significantly reduced and showed obvious demethylation stripes while other groups mainly showed the methylation stripes. The differences of RASSF1A protein expression between AZA, AZA ＋ DDP and AZA ＋MPA groups were not statistical significant （P〈 0. 05）, but the three were higher than model group （P〈0.05）; there was no statistically significant difference respectively in the DDP, MPA, DDP ＋ MPA groups compared with that of model group （P〈0. 05）. In the comparison of apoptosis index, model group was the lowest, followed by the three single medicine groups, and the highest was three combination groups （P〈 0. 05）. Conclusion Demethylation drug 5-Aza-CdR in endometrial cancer treatment has a great potential clinical application value by reversing the abnormal methylation of RASSF1A gene, restoring biological functions of RASSF1A protein and strengthening the efficacy of DDP and MPA.

关 键 词：地西他滨 移植瘤子宫内膜癌 去甲基化 RASSF1A基因 

分 类 号：R737.33[医药卫生—肿瘤]