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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 investigate the effect and mechanism of demethylation drug decitabine on nude mouse endometrial carcinoma xenografts. Methods The nude mice model of endometrial carcinoma was established and divided randomly into decitabine group (AZA group), cisplatin group (DDP group), medroxyprogesterone acetate group (MPA group), AZA + DDP group, AZA + MPA group, DDP + MPA group and model group with 3 rats in each group. The rats in each experimental group were given the corresponding drugs (1μg / g single or 1μg / g each). The model group was given normal saline. Medicine, a total of 8 times administered. The tumor growth of nude mice in each group was observed after treatment. The tumor inhibition rate was calculated after the tumor was removed. The promoter of RASSF1A gene was detected by methylation-specific PCR (MSP), Western blot and TUNEL staining Regional methylation status, protein expression and apoptosis of tumor cells. Results The AZA + DDP group had the highest inhibitory rate of tumor, while AZA group had the lowest inhibitory rate. AZA group, AZA + DDP group, AZA + MPA group RASSF1A gene promoter region methylation decreased significantly, showing a significant non-methylated bands, the remaining group mainly shows methylated bands. The expression of RASSF1A protein in AZA group, AZA + DDP group and AZA + MPA group had no significant difference (P> 0.05), but higher than that in model group (P <0.05) There was no significant difference between DDP + MPA group and model group (P> 0.05). Apoptosis index of the model group <3 single drug group <3 combination group (P <0.05). Conclusion Decitabine reverses the abnormal methylation of RASSF1A promoter region in endometrial carcinoma, restores the biological function of RASSF1A protein, enhances the efficacy of DDP and MPA, and has great potential for the treatment of endometrial cancer Value.