端粒酶逆转录酶线粒体转位促进肝癌细胞干性及耐药的研究

发布时间：2018-05-21 04:13

本文选题：端粒酶逆转录酶 + 线粒体　；参考：《第三军医大学学报》2017年08期

【摘要】：目的探讨人端粒酶逆转录酶(human telomerase reverse enzyme,hTERT)线粒体转位对肝癌细胞干性及耐药的影响。方法采用大剂量顺铂(CDDP)冲击、间歇诱导的方法诱导肝癌细胞HepG2、Huh7耐药,构建耐药细胞株HepG2/CDDP、Huh7/CDDP,CCK-8检测细胞耐药能力;Western blot检测细胞线粒体hTERT表达情况;激光共聚焦显微镜观察细胞线粒体膜电位;流式细胞术检测耐药细胞干性相关蛋白CD133、EpCAM表达情况;克隆形成实验检测细胞克隆形成能力;Western blot检测耐药细胞干性相关因子OCT-4蛋白表达水平变化。结果与亲本细胞HepG2[耐药指数(7.69±0.86)μg/m L及Huh7(7.18±0.35)μg/m L]相比,耐药细胞株对CDDP耐药指数明显增加[HepG2(41.16±0.42)μg/m L,Huh7(33.48±0.33)μg/m L,P0.01],经顺铂(CDDP)诱导耐药细胞线粒体hTERT表达显著升高,线粒体膜电位增强,CD133~+细胞比例[HepG2(1.44±0.41),HepG2/CDDP(23.15±1.55),P0.01]及EpCAM~+细胞比例[HepG2(0.85±0.10),HepG2/CDDP(3.96±0.10),P0.01]增加,克隆形成能力增强,干性相关因子OCT-4蛋白表达水平增加。结论经顺铂诱导的肝癌耐药细胞线粒体hTERT转位显著增加,且具有明显的肿瘤干细胞特征。
[Abstract]:Objective to investigate the effects of mitochondrial translocation of human telomerase reverse transcriptase (telomerase reverse) enzyme hTERT on the dryness and drug resistance of hepatocellular carcinoma cells. Methods HepG _ 2 / CDDPH _ 7 cell line was induced by high dose of cisplatin (CDDP) shock and intermittent induction. The resistant cell line HepG2 / CDDPH7 / CDDPH7 / CDDPCCK-8 was constructed to detect the drug resistance ability of HepG2 / CDDPH7 / CDDPCCK-8 cells. Western blot was used to detect the expression of mitochondrial hTERT. The membrane potential of mitochondria was observed by laser confocal microscopy and the expression of CD133 / EpCAM was detected by flow cytometry. The expression of dry-related factor (OCT-4) protein in drug-resistant cells was detected by Western blot. Results compared with parental cell line HepG2 [drug resistance index 7.69 卤0.86 渭 g / mL and Huh7(7.18 卤0.35 渭 g / mL], the drug resistance index of drug resistant cell line to CDDP increased significantly [HepG2(41.16 卤0.42 渭 g / m L Huh7H7H7 (33.48 卤0.33) 渭 g / m L P0.01], and the expression of mitochondrial hTERT was significantly increased by cisplatin CDDP. The percentage of CD133~ cells increased by mitochondrial membrane potential [HepG2(1.44 卤0.41 + HepG2 / CDDPN 23.15 卤1.55 + P0.01] and the percentage of EpCAM~ cells [HepG2(0.85 卤0.10Hep _ 2 / CDDPN _ (3.96 卤0.10) P _ (0.01)], the clone formation ability was enhanced and the expression of OCT-4 protein was increased. Conclusion the mitochondrial hTERT translocation of drug-resistant hepatoma cells induced by cisplatin is significantly increased and has the characteristics of tumor stem cells.
【作者单位】：第三军医大学新桥医院消化内科;
【分类号】：R735.7

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