地西他滨诱导宫颈癌细胞凋亡分子机制探讨

发布时间：2018-07-07 19:18

本文选题：地西他滨 + Hela细胞　；参考：《中华肿瘤防治杂志》2017年08期

【摘要】：目的地西他滨(decitabine,DAC)具有较好的抗肿瘤活性,其分子机制与DNA去甲基化有关,本研究通过DAC对腺瘤性结肠息肉病(adenomatous polyposiscoli,APC)基因的表达水平和去甲基化的影响,探讨DAC诱导宫颈腺癌Hela细胞凋亡分子机制。方法不同浓度的DAC作用于宫颈腺癌Hela细胞24、36和48h,MTT法检测对宫颈癌细胞的增殖抑制作用,观察DAC对Hela细胞增殖的浓度依赖效应和时间依赖效应。流式细胞术检测DAC对宫颈腺癌Hela细胞凋亡和细胞周期的影响。在DAC作用于宫颈腺癌Hela细胞前后,通过甲基化特异性PCR(Methylation specific PCR,MSP)检测APC基因的甲基化状态;RT-PCR法检测APC基因mRNA表达水平;蛋白质印迹法检测APC蛋白、β-catenin蛋白在胞内及核内表达的变化。结果 DAC对宫颈腺癌Hela细胞增殖抑制具有浓度依赖效应和时间依赖效应,Hela细胞半数抑制浓度(IC50)24、36、48h分别为28.23、7.65和5.64μmol/L。DAC处理后的宫颈腺癌Hela细胞的凋亡率为(73.82±0.11)%,明显高于对照组的(12.41±0.24)%,P0.001。DAC处理Hela细胞后,S期细胞比例(47.82±2.57)%和G2期细胞比例(30.87±2.28)%显著高于空白对照组的S期细胞比例(24.08±0.71)%和G2期细胞比例(2.52±0.84)%,P0.001。DAC处理后,APC基因启动子区域去甲基化状态明显增高,APC mRNA表达量上升,处理前后比较差异有统计学意义,P0.05。DAC处理后,胞内APC蛋白表达上调,而胞内和核内的β-catenin蛋白表达下调,差异均有统计学意义,P0.05。结论 DAC可通过对APC基因的去甲基化作用,上调胞内APC蛋白表达,下调胞内和核内β-catenin表达,诱导宫颈癌细胞凋亡。
[Abstract]:Decitabine DAC has good antitumor activity and its molecular mechanism is related to DNA demethylation. In this study, the effect of DAC on the expression and demethylation of adenomatous polyposis gene was studied. To investigate the molecular mechanism of apoptosis induced by DAC in Hela cells of cervical adenocarcinoma. Methods the inhibitory effects of DAC on the proliferation of cervical carcinoma Hela cells were detected by MTT assay at 24 ~ 36 and 48 h, and the concentration-dependent and time-dependent effects of DAC on Hela cell proliferation were observed. The effect of DAC on apoptosis and cell cycle of Hela cells in cervical adenocarcinoma was detected by flow cytometry. The expression level of APC mRNA was detected by methylation specific PCR, and the expression of APC protein and 尾 -catenin protein in cell and nucleus were detected by Western blotting. Results DAC had concentration-dependent and time-dependent effects on the proliferation inhibition of Hela cells. The apoptotic rate of Hela cells treated with DAC was 28.237.65 and 5.64 渭 mol / L / L DAC for 48 h, respectively, which was significantly higher than that in the control group (73.82 卤0.11). The percentage of S phase cells and G2 phase cells in Hela cells treated with P0.001.DAC were significantly higher than those in blank control group (24.08 卤0.71)% and (2.52 卤0.84) P0.001.DAC, respectively. The percentage of S phase cells and G 2 phase cells in Hela cells treated with P0.001.DAC was significantly higher than that in control group (30.87 卤2.28)%, and the percentage of APC gene promoter region in Hela cells treated with DAC was significantly higher than that in blank control group (24.08 卤0.71)% and (2.52 卤0.84) P0.001.DAC treatment. The expression of APC mRNA increased, After treatment with P0.05.DAC, the expression of APC protein was up-regulated, while the expression of 尾 -catenin was down-regulated in the nucleus and intracellular cells, and the difference was statistically significant (P 0.05). Conclusion DAC can up-regulate the expression of APC protein, down-regulate the expression of 尾 -catenin and induce apoptosis of cervical cancer cells by demethylation of APC gene.
【作者单位】：南华大学附属南华医院妇产科;
【分类号】：R737.33

【相似文献】