今日药学

2020, v.30(10) 668-672+682

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DDX11-AS1在食管癌细胞对紫杉醇耐药中的作用及机制研究
Study of the Role and Mechanism of DDX11-AS1 in Paclitaxel Resistance of Esophageal Cancer Cells

陈晨;廖巍;傅永锦;张盛奇;古淑仪;吴钦水;陈云;许诚城;张述耀;
CHEN Chen;LIAO Wei;FU Yongjin;ZHANG Shengqi;GU Shuyi;WU Qinshui;CHEN Yun;XU Chengcheng;ZHANG Shuyao;Tumor Hospital of Shantou University Medical College;Guangzhou Red Cross Hospital of Jinan University;Dafeng Hospital of Shantou;Shantou University Medical College;

摘要(Abstract):

目的探究长链非编码RNA DDX11-AS1与食管癌对紫杉醇耐药的关系。方法收集汕头大学医学院附属肿瘤医院2017年5月~2018年10月间食管癌连续性手术切除标本40例。(1)耐药株R-EC109敲减拓扑异构酶IIα(TOP2A),亲本细胞EC109中过表达TOP2A,检测两种细胞对紫杉醇敏感性变化。(2)双荧光素酶试验检测DDX11-AS1和转录因子TAF1对TOP2A启动子活性的影响。(3)荧光原位杂交试验定位DDX11-AS1,CHIP和RIP试验检测TOP2A启动子与TAF1以及DDX11-AS1与TAF1之间的联系。(4) PCR和免疫组化检测癌及癌旁组织中DDX11-AS1、TOP2A和TAF1的差异,分析DDX11-AS1与TOP2A的相关性。结果 (1) R-EC109中TOP2A高表达,敲减TOP2A可提高R-EC109对紫杉醇的敏感性。(2)双荧光素酶报告基因试验表明,DDX11-AS1和TAF1提高TOP2A启动子转录活性。(3) DDX11-AS1主要定位于核内,TOP2A启动子和TAF1以及DDX11-AS1和TAF1存在相互作用。(4) DDX11-AS1、TOP2A和TAF1高表达,DDX11-AS1与TOP2A的表达量呈正相关。结论 DDX11-AS1通过TAF1促进TOP2A的转录,增强食管癌细胞对紫杉醇耐药。
OBJECTIVE To investigate the relationship between lncRNA DDX11-AS1 and paclitaxel resistance in esophageal cancer. METHODS From May 2017 to October 2018,40 consecutive surgical specimens of esophageal cancer were collected from tumor hospital of Shantou university medical college.( 1) TOP2 A was knocked down by R-EC109,and TOP2 A was overexpressed in its parent cell EC109. The sensitivity of the two cells to paclitaxel were detected.( 2) The effects of DDX11-AS1 and TAF1 on the activity of TOP2 A promoter were detected by double luciferase assay.( 3) DDX11-AS1 was located by FISH assays. ChIP and Rip assays were used to detect the association between TOP2 A promoter and TAF1,and DDX11-AS1 with TAF1.( 4) The differences of DDX11-AS1,TOP2 A and TAF1 in cancer and adjacent tissues were detected by PCR and immunohistochemistry,and the correlation between DDX11-AS1 and TOP2 A were analyzed. RESULTS( 1) TOP2 A was highly expressed in R-EC109. Knockdown of TOP2 A could increase the sensitivity of R-EC109 to paclitaxel.( 2) Double luciferase reporter gene test showed that DDX11-AS1 and TAF1 increased the transcriptional activity of TOP2 A promoter.( 3) DDX11-AS1 is mainly located in the nucleus. TOP2 A promoter interacts with TAF1 and DDX11-AS1 interacts with TAF1.( 4) The expression of DDX11-AS1,TOP2 A and TAF1 were high,and the expression of DDX11-AS1 was positively correlated with that of TOP2 A. CONCLUSION DDX11-AS1 promotes the transcription of TOP2 A through TAF1 and enhances the resistance of esophageal cancer cells to paclitaxel.

关键词(KeyWords): 食管癌;紫杉醇;DDX11-AS1;耐药
esophageal cancer;paclitaxel;DDX11-AS1;drug resistance

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作者(Author): 陈晨;廖巍;傅永锦;张盛奇;古淑仪;吴钦水;陈云;许诚城;张述耀;
CHEN Chen;LIAO Wei;FU Yongjin;ZHANG Shengqi;GU Shuyi;WU Qinshui;CHEN Yun;XU Chengcheng;ZHANG Shuyao;Tumor Hospital of Shantou University Medical College;Guangzhou Red Cross Hospital of Jinan University;Dafeng Hospital of Shantou;Shantou University Medical College;

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