MiR-92b-3p通过靶向RAD21增强非小细胞肺癌对顺铂的化学敏感性  

Enhancement of chemosensitivity to cisplatin in non-small cell lung cancer through MiR-92b-3p by targeting RAD21

作　　者：潘有光[1] 傅文凡 莫益俊 赵健 陈岗东[1] PAN Youguang;FU Wenfan;MO Yijun;ZHAO Jian;CHEN Gangdong(The Third Affiliated Hospital of Guangzhou Medical University,Guangzhou 510150,China)

机构地区：[1]广州医科大学附属第三医院,广州510150 [2]广州医科大学附属肿瘤医院,广州510095 [3]南方医科大学深圳医院,广东深圳518000

出　　处：《实用医学杂志》2020年第14期1897-1902,共6页The Journal of Practical Medicine

基　　金：广东省医学科学技术研究基金项目(编号:B2018186)。

摘　　要：目的检测miR-92b-3p在非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞株A549及其顺铂耐药株A549/DDP中的表达,探讨其在顺铂耐药中的作用机制。方法应用实时定量反转录聚合酶联反应(qPCR)检测miR-92b-3p在A549/DDP及A549中的表达;CCK8法分别检测各细胞株转染miR-92b-3p mimics或inhibitor后对DDP敏感度的改变;生物信息学预测及双荧光素酶报告基因验证miR-92b-3p和靶基因的关系。qPCR和Western Blot检测RAD21基因及蛋白的表达。结果 miR-92b-3p在A549/DDP细胞株中呈现低表达(P <0.05),miR-92b-3p mimics转染A549/DDP细胞株后,DDP对转染细胞株的半数抑制浓度(half inhibition concentration,IC50)显著低于阴性对照组(P <0.05);同时该细胞株的RAD21的mRNA及蛋白表达水平明显低于阴性对照组(P <0.05)。反之,在A549细胞株中转染miR-92b-3p inhibitor后,得出相反的结论。结论 miR-92b-3p在A549/DDP细胞株低表达,其通过调控RAD21 mRNA及蛋白的表达而逆转A549/DDP细胞对DDP的耐药。Objective To investigate the expression of mi R-92b-3p in non-small cell lung cancer(NSCLC) cell line A549 and its cisplatin resistant line A549/DDP and to explore its mechanism in cisplatinresistance.MethodsThe expression of mi R-92b-3p in A549/DDP cell lines and A549 cell lines was detected byreal-time quantitative reverse transcription polymerase(q PCR). CCK8 method was used to detect the change of thesensitivity to DDP after the cell line transfect mi R-92b-3p mimics or inhibitor respectively and bioinformatics topredict the target gene of mi R-92b-3p. The relationship between mi R-92b-3p and the target gene was verified bydual luciferase reporter and the expression of RAD21 gene and its protein was detected by q PCR and Western Blot.Resultsmi R-92b-3p showed low expression in A549/DDP cell lines(P < 0.05). After mi R-92b-3p mimics weretransfected into A549/DDP cell lines,half inhibition concentration(IC50)of DDP was significantly lower than thatof negative control group(P < 0.05). Meanwhile,the m RNA and protein expression levels of RAD21 in this cellline were significantly lower than those in negative control group(P < 0.05). Conversely,after mi R-92b-3p inhibitorwere transfected into A549 cell line,the opposite conclusion was obtained.Conclusionmi R-92b-3p is underex-pressed in A549/DDP cell lines,and it reverses drug resistance of A549/DDP cells to DDP by regulating theexpression of RAD21 m RNA and protein.

关 键 词：非小细胞肺癌 miR-92b-3p 化疗耐药 顺铂 RAD21 

分 类 号：R734.2[医药卫生—肿瘤]