Smac/DIABLO增强子宫内膜癌细胞对紫杉醇化疗敏感性的机制研究  

Mechanism of Smac/DIABLO enhances the paclitaxel chemotherapy in endometrial carcinoma

作　　者：王晓华 韩冰[1] 张玉娟[1] Wang Xiaohua;Han Bing;Zhang Yujuan(Department of Gynaecology,Affiliated Hospital of Chengde Medical Unversity,Hebei Chengde 067000,China)

机构地区：[1]承德医学院附属医院妇一科

出　　处：《现代肿瘤医学》2019年第18期3194-3198,共5页Journal of Modern Oncology

基　　金：河北省承德市科学技术研究与发展计划项目(编号:201804A026)

摘　　要：目的:研究上调Smac/DIABLO基因在人子宫内膜癌Ishikawa细胞中的表达,对紫杉醇化疗增敏方面的作用及可能机制。方法:构建Smac/DIABLO的过表达质粒pcDNA3.1-Smac,应用脂质体法转染人子宫内膜癌Ishikawa细胞,实验分为过表达组、空载体对照组和空白对照组。转染48 h后,应用qRT-PCR和Western blot法检测Smac/DIABLO mRNA及蛋白表达情况。将转染pcDNA3.1-Smac的Ishikawa细胞暴露于终浓度为0.1μmol/L紫杉醇中,实验分为pcDNA3.1-Smac+紫杉醇组、紫杉醇组和空白对照组,CCK-8法检测暴露紫杉醇24,48,72,96h,3组细胞增殖能力的变化。应用Western blot检测Ishikawa细胞过表达Smac/DIABLO并暴露于紫杉醇后,Caspase-3,Caspase-7,Caspase-9蛋白的表达情况。结果:转染48 h后,过表达组较空载体对照组、空白对照组,Smac/DIABLO mRNA及蛋白相对表达量均明显增加(F=152.056,P=0.002;F=697.953,P=0.001)。CCK-8实验结果显示,过表达Smac/DIABLO并暴露于紫杉醇后,pcDNA3.1-Smac+紫杉醇组较紫杉醇组和空白对照组,细胞增殖能力受到抑制(P<0.05)。Western blot检测到pcDNA3.1-Smac+紫杉醇组Caspase-3,Caspase-7,Caspase-9蛋白的相对表达量比较另外两组明显增加(FCaspase-3=434.277,FCaspase-7=392.401,FCaspase-9=88.936,P<0.05)。结论:上调Smac/DIABLO表达可增强Ishikawa细胞对紫杉醇的化疗敏感性,其机制可能通过线粒体和死亡受体两条途径,解除凋亡抑制蛋白家族对Caspase的抑制作用。Objective:To study the up-regulation of Smac/DIABLO gene expression in human endometrial carcinoma Ishikawa cells,the effect and possible mechanism of paclitaxel chemosensitization.Methods:The overexpression plasmid pcDNA3.1-Smac of Smac/DIABLO was constructed with pcDNA3.1(+) as the cloning vector.Ishikawa cells were transfected with liposomes.The experiment was divided into overexpression group,empty vector control group and blank control group.The expression of Smac/DIABLO mRNA and protein was detected by qRT-PCR and Western blot after 48 h transfection. Ishikawa cells transfected with pcD NA3. 1-Smac were exposed to a final concentration of 0. 1 μmol/L paclitaxel. The experiment was divided into pcD NA3. 1-Smac + paclitaxel group,paclitaxel group and blank control group. CCK-8 method was used to detect the changes of cell proliferation ability in cells exposed to paclitaxel at 24,48,72 and 96 h. The expression of Caspase-3,Caspase-7,Caspase-9 protein was detected by Western blot after exposure to paclitaxel. Results: After transfection for 48 h,the expression of Smac/DIABLO mRNA and protein in overexpression group was significantly higher than that in empty vector control group and blank control group( F = 152. 056,P = 0. 002;F = 697. 953,P = 0. 001). CCK-8 results show that overexpression of Smac/DIABLO and exposure to paclitaxel,compared with paclitaxel group and blank control group,the cell proliferation ability of pcD NA3. 1-Smac + paclitaxel group was significantly inhibited( P < 0. 05). The relative expression of Caspase-3,Caspase-7,Caspase-9 protein in pcD NA3. 1-Smac + paclitaxel group was significantly higher than that in the other two groups( FCaspase-3= 434. 277,FCaspase-7= 392. 401,FCaspase-9= 88. 936,P < 0. 05). Conclusion:Up-regulation of Smac/DIABLO expression enhances the chemosensitivity of Ishikawa cells to paclitaxel,and its mechanism may be achieved by both the mitochondrial and death receptors,and the inhibition of Caspase by the inhibitory protein family.

关 键 词：子宫内膜癌 SMAC/DIABLO 过表达 紫杉醇 化疗敏感性 

分 类 号：R737.33[医药卫生—肿瘤]