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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P<0.01)。促进凋亡蛋白Bax和抑制凋亡蛋白Bcl2的比值亦较明显增加(P<0.01)。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。
Objective: To investigate the role of recombinant human programmed cell death protein 5 (rhPDCD5) in paclitaxel chemotherapy in endometrial cancer cells. Methods: KLE cells were treated with recombinant human rh PDCD5 (20μg / ml) after KLE cells were cultured in endometrial carcinoma, and then treated with 0,1.0,5.0,10.0 and 50μmol / L paclitaxel (PTX) for 24 h or After treated with 10μmol / L PTX for 0, 12, 24 and 48 hours, the total RNA and protein of the cells were extracted. The proliferation of KLE cells was detected by CCK method. The apoptosis of KLE cells was detected by flow cytometry and KLE cells by real- PDCD5mRNA expression levels, real-time quantitative PCR or Western blotting determination of apoptotic-related genes Bax, Bcl2, caspase-3 mRNA or protein levels. Results: The promoting effect of PTX on PDCD5 expression was dose-dependent and time-dependent. The optimal concentration of PTX was 10 μmol / L and the best action time was 24 h. rh PDCD5 significantly enhanced the inhibitory effect of paclitaxel on KLE cells. The results of CCK assay, flow cytometry and Western blotting showed that the proliferation and apoptosis rates of KLE cells were significantly increased and the expression of pro-caspase 3 was significantly increased (P <0.01, P <0.01) ). The ratio of promoting apoptosis protein Bax and inhibiting apoptosis protein Bcl2 also increased obviously (P <0.01). CONCLUSION: rhPDCD5 can synergize with PTX to inhibit the proliferation of KLE cells and promote the apoptosis of endometrial carcinoma cells. It can significantly enhance the drug sensitivity of KLE cells to PTX.