紫杉醇靶向Hippo信号通路抑制食管鳞癌干细胞干性的机制研究  

Paclitaxel Governs Stemness of Esophageal Squamous Cell Carcinoma Stem Cells Through Hippo Signal Pathway

作　　者：宿伟鹏[1] 张洋[1] 张宋安[1] 刘攀[1] 赵化荣[1] Su Weipeng;Zhang Yang;Zhang Songan(Oncology Department,The First Affiliated Hospital of Xinjiang Medical University,Xinjiang 830054,China)

机构地区：[1]新疆医科大学第一附属医院,乌鲁木齐830054

出　　处：《医学研究杂志》2020年第1期76-80,75,共6页Journal of Medical Research

基　　金：新疆维吾尔自治区自然科学基金资助项目(2016D01C263)。

摘　　要：目的探究紫杉醇靶向Hippo信号通路抑制食管鳞癌干细胞干性的作用机制。方法流式细胞术分离TE-13细胞系中侧群细胞,特异性筛选分离食管鳞癌干细胞(ESCC-CSCs),经免疫荧光鉴定后将细胞分为对照组及实验组,对照组细胞不做任何处理,实验组细胞给予1.25μg/ml紫杉醇。CCK-8、克隆形成检测细胞体外增殖活力的变化,成球实验检测细胞自我复制及更新能力的变化,QRT-PCR检测细胞内相关分子mRNA表达变化,Western blot法检测细胞内相关蛋白表达变化。结果TE-13细胞中分选得到29.3%ESCC-CSCs细胞,给予紫杉醇诱导后,细胞体外增殖能力受到显著抑制(P=0.000),克隆形成数量显著少于对照组(P<0.01),成球实验结果表明实验组体外成球个数及球体直径与对照组比较,差异有统计学意义(P<0.01);Western blot法检测结果表明,紫杉醇治疗后ESCC-CSCs细胞内TAZ、CD44、SOX2蛋白表达降低,p-TAZ蛋白含量升高,与对照组比较,差异有统计学意义;qRT-PCR结果显示实验组细胞内CD44、SOX2 mRNA表达降低,而TAZ mRNA表达与对照组比较,差异无统计学意义。结论紫杉醇可靶向Hippo信号通路中TAZ、SOX2、CD44等关键分子参与调控ESCC-CSCs细胞干性。Objective To investigate the mechanism of paclitaxel governs stemness of ESCC-CSCs through Hippo signal pathway.Methods The cancer stem cells were isolated from the esophageal squamous cell carcinoma cell line TE-13 by flowcytometry.Immunofluorescence was used to identify the stem cells for further research.ESCC-CSCs were divided into the control group and the experimental group.The experimental group was given 1.25μg/ml paclitaxel while the control group were treated with nothing.Colony formation and CCK-8 assays were used to detect the changes in cell proliferation after treated with paclitaxel.Sphere formation assays was used to detect the changes in self-replication and renewal abilities.qRT-PCR was used to detect the expression of relative mRNAs.Western blot was used to detect the expression of relative proteins in ESCC-CSCs.Results 29.3%TE-13 cells were sorted as ESCC-CSCs.After treated with paclitaxel,the proliferation of ESCC-CSCs was significantly suppressed than the control group(P=0.000).The number of clones was significantly less than that of the control group(P<0.01).The number and the size of spheres in the experimental group were significantly different from those in the control group(P<0.01).The expression of TAZ,CD44,SOX2 proteins were suppressed while the expression of p-TAZ was significantly increased in the experimental group.QRT-PCR showed that the expression of SOX2 and CD44 mRNA in the experimental group were apparently decreased than in the control group.However,there was no difference in TAZ mRNA expression between the two groups.Conclusion Paclitaxel can target key molecules,such as TAZ,SOX2 and CD44,to regulate the stemness of ESCC-CSCs.

关 键 词：紫杉醇 食管鳞癌 干细胞 HIPPO 

分 类 号：R735[医药卫生—肿瘤]