rhPDCD5促进子宫内膜癌KLE细胞对紫杉醇的药物敏感性

发布时间：2018-02-26 20:28

本文关键词： 重组人程序性细胞死亡蛋白子宫内膜癌紫杉醇耐药性敏感性　出处：《中国肿瘤生物治疗杂志》2017年10期 　论文类型：期刊论文

【摘要】：目的:探讨在子宫内膜癌细胞中重组人程序性细胞死亡蛋白5(recombinant human programmed cell death protein 5,rhPDCD5)对紫杉醇化疗的促进作用。方法:子宫内膜癌KLE细胞培养完成后,通过重组人rh PDCD5(20μg/ml)处理KLE细胞,再分别以0、1.0、5.0、10.0、50μmol/L紫杉醇(paclitaxel,PTX)处理24 h或以10μmol/L PTX处理0、12、24、48 h,提取细胞总RNA及蛋白后,CCK法检测KLE细胞的增殖情况,流式细胞术检测KLE细胞凋亡情况,实时定量PCR检测KLE细胞中PDCD5mRNA的表达量,实时定量PCR或Western blotting测定凋亡相关基因的Bax、Bcl2、caspase-3 mRNA或蛋白水平的变化。结果:PTX对PDCD5表达的促进作用具有剂量依赖性和时间依赖性;PTX的最佳作用浓度为10μmol/L,最佳作用时间为24 h。rh PDCD5明显增强紫杉醇对KLE细胞的抑制作用。CCK实验、流式细胞术及Western blotting检测显示:PTX+rhPDCD5联合处理组KLE细胞的增殖抑制率和凋亡率均较PTX组明显增加、pro-caspase 3的表达量明显增加(均P0.01)。促进凋亡蛋白Bax和抑制凋亡蛋白Bcl2的比值亦较明显增加(P0.01)。结论:rhPDCD5可协同PTX抑制子宫内膜癌KLE细胞的增殖、促进细胞的凋亡,可明显增强KLE细胞对PTX的药物敏感性。
[Abstract]:Objective: to investigate the effect of recombinant human programmed cell death protein 5 (rhPDCD5) on paclitaxel chemotherapy in endometrial cancer cells. Methods: KLE cells were treated with recombinant human rhPDCD5 20 渭 g / ml after the completion of KLE cell culture. The cell total RNA and protein were extracted for 24 h or 10 渭 mol/L PTX for 24 h, respectively. The proliferation of KLE cells was detected by flow cytometry. The apoptosis of KLE cells was detected by flow cytometry, and the expression of PDCD5mRNA in KLE cells was detected by real time quantitative PCR. Real time quantitative PCR or Western blotting was used to detect the changes of Baxia Bcl2 caspase-3 mRNA or protein in apoptosis-related genes. Results the optimal concentration of PDCD5 expression was 10 渭 mol / L and the optimal time was 10 渭 mol / L for the promotion of PDCD5 expression by 1: PTX in a dose and time dependent manner. The inhibitory effect of paclitaxel on KLE cells was significantly enhanced by 24 h. Rh PDCD5. The results of flow cytometry and Western blotting analysis showed that the proliferation inhibition rate and apoptosis rate of KLE cells treated with WPTX rhPDCD5 were significantly higher than those of PTX group (all P 0.01). The ratio of Bax to Bcl2 was significantly higher than that of PTX group. Conclusion: rhPDCD5 can inhibit the proliferation of endometrial carcinoma KLE cells in combination with PTX. The drug sensitivity of KLE cells to PTX was significantly enhanced by promoting cell apoptosis.
【作者单位】：重庆市中医院肿瘤科;
【分类号】：R737.33

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