rhPDCD5促进子宫内膜癌KLE细胞对紫杉醇的药物敏感性  

rhPDCD5 enhances susceptibility of endometrial cancer KLE cells to paclitaxel

作　　者：汪宇宏[1] 王怀碧[1] 邱敏[1] 

机构地区：[1]重庆市中医院肿瘤科,重庆400020

出　　处：《中国肿瘤生物治疗杂志》2017年第10期1101-1106,共6页Chinese Journal of Cancer Biotherapy

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 explore the promoting effect of rhPDCD5 （recombinant human programmed cell death protein 5） on paclitaxel （PTX） chemotherapy in endometrial carcinoma cells. Methods： After routine culture, endo- metrial carcinoma KLE cells were treated with rhPDCD5 （20 μg/ml）, and then treated with PTX at different concen- trations （0, 1.0, 5.0, 10.0 and 50 μmol/L） for 24 hours or 10 μmol/L PTX for different time periods （0,12,24,48 h）. The proliferation of KLE cells was determined by CCK, the apoptosis was examined by Flow cytometry, the PDCD5 mRNA was determined by Real-time quantitative （qPCR）, and the changes of apoptosis-related genes （Bax, Bcl2, caspase 3） at protein and mRNA level were detected by western blotting and q-PCR, respectively. Results： The promoting effect of PTX on PDCD5 expression was dose- and time- dependent. The optimal concentration of PTX was 10 μmol/L and the best treatment duration was 24 hours, rhPDCD5 significantly promoted the inhibitory effect of PTX on KLE cells. CCK-8 assay and Flow cytometry confirmed that the proliferation inhibitory rate and apoptosis rate were significantly enhanced in PTX＋rhPDCD5 group, compared with PTX group （P〈0.01）; In the presence of PTX and rhPDCD5, the expression level of pro-caspase 3 was significantly increased （P〈0.01）; more- over, the ratio of Bax to Bcl2 was significantly higher in PTX＋rhPDCD5 group than that in other groups （P〈0.01）. Conclusion： rhPDCD5 can enhance the inhibitory effect of PTX on proliferation of KLE cells, promote cell apopto- sis and significantly enhance the PTX susceptibility of KLE cells.

关 键 词：重组人程序性细胞死亡蛋白5 子宫内膜癌 紫杉醇 耐药性 敏感性 

分 类 号：R737.33[医药卫生—肿瘤] R730.5[医药卫生—临床医学]