敲低CDCA8基因表达对膀胱癌5637细胞吡柔比星敏感性的影响  

Effect of CDCA8 gene expression knockdown on the sensitivity of pirarubicin in bladder cancer 5637 cells

作　　者：高鑫 陈美 王新彤 刘明月 张淑芳[2] GAO Xin;CHEN Mei;WANG Xin-tong;LIU Ming-yue;ZHANG Shu-fang(Clinical Laboratory,The Fourth Affiliated Hospital of Jishou University/The First People′s Hospital of Huaihua,Huaihua 418000,Hunan,China;Central Laboratory,Affiliated Haikou Hospital of Xiangya Medical College,Central South University,Haikou 570208,Hainan,China;Jishou University School of Medicine,Jishou 416000,Hunan,China)

机构地区：[1]吉首大学第四临床学院怀化市第一人民医院检验科,怀化418000 [2]中南大学湘雅医学院附属海口医院中心实验室,海口570208 [3]吉首大学医学院,吉首416000

出　　处：《医学研究生学报》2022年第6期592-597,共6页Journal of Medical Postgraduates

基　　金：湖南省自然科学基金(2021JJ40427);吉首大学校级科研项目(Jdlc2021)。

摘　　要：目的膀胱癌具有较高的耐药和复发率,研究其分子机制有助于解决临床耐药问题。文中旨在探索CDCA8在膀胱癌5637细胞中的表达与吡柔比星(THP)敏感性的相关性。方法收集2018年1月至2020年12月中南大学湘雅医学院附属海口医院膀胱癌组织患者的临床信息。从TCGA数据库中获取CDCA8的mRNA表达数据及患者化疗信息,同时进行免疫组化实验,分析CDCA8的表达与患者化疗敏感性的关系。针对CDCA8基因构建RNA干扰慢病毒,将膀胱癌5637细胞分为空白组(未作处理)、阴性对照组(转染不含CDCA8靶序列的慢病毒)和实验组(转染含CDCA8靶序列的慢病毒,敲低CDCA8),检测CDCA8基因及蛋白的表达;检测敲低CDCA8对膀胱癌细胞THP敏感性的影响;Celigo检测5637细胞吡柔比星IC50。结果膀胱癌化疗者CDCA8表达(10.369±1.131)较非化疗者(9.250±0.892)显著高表达(P=0.012)。qRT-PCT实验结果显示实验组CDCA8的mRNA表达量(0.069±0.004)较空白组(1.087±0.075)、阴性对照组(1.001±0.054)显著降低(P<0.001)。免疫印迹实验结果所示,实验组CDCA8蛋白表达量较空白组、阴性对照组明显降低(P=0.003)。实验组的IC50明显低于空白组、阴性对照组(P<0.05),细胞抑制率显著高于其他两组(P<0.05)。结论敲低CDCA8表达后增强了膀胱癌5637细胞对THP的敏感性,为后续研究膀胱癌THP耐药机制奠定了研究基础。Objective Bladder cancer has a high rate of drug resistance and recurrence.The study of its molecular mechanism is helpful to solve the problem of clinical drug resistance.This study aims to explore the correlation between CDCA8 expression and the sensitivity of pirarubicin(THP)in bladder cancer 5637 cells.Methods Clinical information of patients with bladder cancer tissue in Haikou Hospital affiliated to Xiangya School of Medcine,Central South University from January 2018 to December 2020 was collected.CDCA8 mRNA expression data and chemotherapy information of patients were obtained from TCGA database,and immunohistochemical experiments were performed to analyze the relationship between CDCA8 expression and chemotherapy sensitivity of patients.The RNA interference lentivirus targeting CDCA8 was constructed,and bladder cancer 5637 cells were divided into blank group(untreated),negative control group(transfected with lentivirus without CDCA8 target sequence)and experimental group(transfected with lentivirus containing CDCA8 target sequence and knockdown CDCA8).The CDCA8 gene and protein expression,and the effect of CDCA8 knockdown on THP sensitivity of bladder cancer cells were detected;pirarubicin IC50 in bladder cancer 5637 cells was detected by Celigo.Results CDCA8 expression was significantly higher in bladder cancer chemotherapy patients(10.369±1.131)than in non-chemotherapy patients(9.250±0.892)(P=0.012).QRT PCT results showed that CDCA8 mRNA expression in experimental group(0.069±0.004)was significantly lower than that in blank group(1.087±0.075)and negative control group(1.001±0.054)(P<0.001).Western blot results showed that the expression of CDCA8 protein in experimental group was significantly lower than that in blank group and negative control group(P=0.003).The IC50 of the experimental group was significantly lower than that of the blank group and the negative control group(P<0.05),and the cell inhibition rate was significantly higher than that of the other two groups(P<0.05).Conclusion Knockdown

关 键 词：膀胱癌 吡柔比星 CDCA8 耐药 生物信息学 

分 类 号：R737.14[医药卫生—肿瘤]