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基础研究

利用SmartFlare技术进行胶质瘤细胞系U87中Twist1hi亚群分选及其对顺铂耐药性研究

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  • 1.首都医科大学附属北京天坛医院临床医学研究实验室,北京 100050;2.首都医科大学附属北京天坛医院药剂科,北京 100050;3.首都医科大学附属北京天坛医院检验科,北京 100050

收稿日期: 2013-12-10

  修回日期: 2014-04-22

  网络出版日期: 2015-01-25

基金资助

 

首都医科大学附属北京天坛医院青年科研基金(2013年度);脑肿瘤北京市重点实验室2014年开放课题(2014NZLY01);北京市自然科学基金(7132094);北京市卫生系统高层次卫生技术人才队伍建设专项经费(2011-3-038)

Twist1 Confers Cisplatin Resistance in Glioma U87 Cells by Inhibiting Cisplatin-induced Apoptosis

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  • Core Laboratory for Clinical Medical Research, Beijing Tiantan Hospital, Capital Medical University, Beijing 100050, China

Received date: 2013-12-10

  Revised date: 2014-04-22

  Online published: 2015-01-25

摘要

目的 利用特异性SmartFlareTM RNA检测探针在人脑胶质瘤细胞系U87中分选Twist1hi-U87细胞亚群,并研究其对顺铂的耐药性。方法 用SmartFlareTM RNA探针对Twist1hi-U87细胞亚群进行分选。以未分选细胞作为对照。CCK8法检测Twist1hi-U87亚群和亲代U87细胞对顺铂的化疗敏感性。流式细胞仪检测顺铂作用后细胞凋亡情况。结果 根据细胞核内Twist1的表达量,流式细胞分选所得Twist1hi-U87亚群约占亲代U87细胞的57.9%~69.8%。CCK8结果显示,加药后24h 和48h,Twist1hi-U87亚群对顺铂的耐药性均高于亲代U87细胞(P<0.05)。经顺铂处理后,Twist1hi-U87细胞凋亡率(14.07% ± 0.96%)明显低于U87细胞(20.4% ± 1.29%)(P<0.05)。结论 Twist1基因高表达可以抑制胶质瘤细胞凋亡,进而降低顺铂的杀伤作用。

关键词: 脑胶质瘤; Twist1; 顺铂; 耐药

本文引用格式

董成亚1,石晓旭2,贺文艳1,李昊文1,刘 丽 1,王雅杰1,3 . 利用SmartFlare技术进行胶质瘤细胞系U87中Twist1hi亚群分选及其对顺铂耐药性研究[J]. 标记免疫分析与临床, 2015 , 22(1) : 41 . DOI: 10.11748/bjmy.issn.1006-1703.2015.01.015

Abstract

Objective Twist1 is a transcription factor that is highly expressed in many human cancers and its expression is related to chemotherapy resistance. In this study, we aimed to enrich for cells with a high-level expression of Twist1 from human glioma cell line U87 and investigate their sensitivity to the chemotherapeutic agent cisplatin. Methods Twist1hi-U87 subsets were sorted through flow cytometry using Twist1 specific SmartFlareTM RNA probe. Unsorted U87 cells were served as control. Different concentrations of cisplatin were administered to the cells. Cell viability was measured by CCK-8 assay and apoptosis was detected by flow cytometry. Results The percentage of Twist1hi-U87 subpopulations in unsorted parental cells was 57.9%~69.8%. CCK-8 assay showed that Twist1hi-U87 subsets were more resistant to cisplatin than U87 cells at 24h, 48h (P<0.05). In addition, Twist1hi-U87 subsets had a lower rate of apoptosis than did the parental U87 cells (14.07% ± 0.96% vs 20.4% ± 1.29%) (P<0.05). Conclusion Twist1 is involved in cisplain resistance by inhibiting cisplatin-induced apoptosis.
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