沉默微小RNA-16通过激活核因子-κB通路介导肝癌细胞化疗耐药  

Silencing microRNA - 16 induces drug resistance of hepatoma cell lines via nuclear factor - KBpathway

作　　者：黄颖滨 陈光宇 汪洋[1] 何睿[1] 杜军[2] 焦兴元[1] 邰强[1] Huang Yingbin;Chen Guangyu;Wang Yang;He Rui;Du Jun;Jiao Xingyuan(Tai Qiang Organ Transplantation Center,the First Affiliated Hospital,Sun Yat-Sen University,Guangzhou 510080,China;School of Pharmaceutical Sciences,Sun Yat-sen University,Guangzhou 510006,China)

机构地区：[1]中山大学附属第一医院器官移植中心,广州510080 [2]中山大学药学院,广州510006

出　　处：《中华实验外科杂志》2018年第9期1654-1656,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81270531）;广东省自然科学基金（S2013010015475）

摘　　要：目的探讨沉默SMMC-7721中微小RNA（miRNA，miR）-16的表达对化疗耐药的影响及机制。方法采用慢病毒载体构建稳定细胞株，应用实时定量反转录聚合酶链反应（RT-qPCR）检测miR-16的表达及Westernblot检测靶基因KB激酶抑制剂B（IKBKB）的表达确定构建结果。采用Westernblot及噻唑蓝（MTF）比色法检测各细胞株对紫杉醇的敏感性及耐药机制。结果7721-miRl6组的miR-16表达量显著高于SMMC-7721组及7721．mock组（6．06±0．18比1．00±O．13比0．87±0．03，P=0．000）。7721-shmiR组靶基因IKBKB蛋白表达量是7721-shmock组的25．73倍（P=0．000）。7721-shmiRl6细胞株中多药耐药蛋白1（MDRl）表达显著增加，对紫杉醇的半数抑制浓度（IC50）为4．903txmol／L，是对照组（0．461μmol／L）的10．64倍（P=0．000）。Westernblot结果显示沉默miR-16主要通过激活细胞核因子-κB（NF．KB）通路导致MDRl表达增加。结论沉默SMMC-7721中miR-16的表达，能显著增加IKBKB的表达，从而导致SMMC-7721细胞对紫杉醇耐药。Objective Study on the mechanism of sliencing microRNA （miRNA, miR） - 16 on chemoresistance in SMMC -7721. Methods The lentiviral vectors were used to construct stable cell lines. To determine whether constructed successfully, the expression of miR - 16 was detected by real - time reverse transcription - polymerase chain reaction （ RT - qPCR） and inhibitor of kappaB kinase beta （IKBKB）, the target gene of miR -16, was detected by Western blotting. The sensitivity and resistance mechanism of the stable cell lines to paclitaxel were detected by Western blotting and thiazole blue （MTF）. Results The expression of miR - 16 in 7721 - miR16 group was significantly higher than SMMC-7721 and 7721- mock groups （4.63±1.32 vs. 1.00±0.30 vs. 0.93±0.37, P =0.000）. The expression of IKBKB in 7721 - shmiR16 was 25.73 times than the controls （P = 0. 000）. The expres- sion of muhidrug resistance protein 1 （ MDR1 ） in 7721 - shmiR16 was increased and the half maximal in- hibitory concentration （IC50） to paclitaxel was 4. 903μmol/L, 10. 64 times than the control group （0. 461 μmol/L） （P = 0. 000）. The results of western showed that silencing miR- 16 induced resistance mainly through activating nuclear factor - kappaB （ NF - KB） pathway. Conclusion Silencing miR - 16 in SMMC -7721 significantly increases IKBKB expression leadind paclitaxel resistance.

关 键 词：肝癌 微小RNA-16 紫杉醇 耐药 κB激酶抑制剂β 

分 类 号：R735.7[医药卫生—肿瘤]