miR374b通过调节ABCG2逆转小细胞肺癌对伊利替康的耐药  

MiR-374b activates the sensitivity of lung cancer cells to Irinotecan via regulating ABCG2 expression

作　　者：曹亮[1] 张长洪[1] 潘磊 支学军[1] 姬泽萱 CAO Liang;ZHANG Chang-hong;PAN Lei;ZHI Xue-jun;JI Ze-xuan(First Affiliated Hospital of Hebei North University Respiratory Medicine,Zhangjiakou075000,P.R.Chin)

机构地区：[1]河北北方学院附属第一医院呼吸内科

出　　处：《中华肿瘤防治杂志》2019年第19期1436-1442,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：河北省卫生厅科研基金(20160035);2018年度市级科技计划项目(1821150H)

摘　　要：目的 ABCG2是促进细胞耐药性的基因,其高表达与肿瘤细胞耐药性的形成密切相关。miR-374b可以抑制肿瘤细胞活力,促进凋亡,是常见的抑癌基因。本研究旨在探讨miR-374b与ABCG2的靶向关系,及其在小细胞肺癌(small cell lung cancer,SCLC)耐药性形成过程中的作用。方法选取2017-05-11-2017-12-17河北北方学院附属第一医院一线治疗失败的SCLC组织20例以及癌旁组织20例。所有实验数据均为3次平行实验均值。采用伊立替康(Irinotecan,CPT-11)诱导产生耐药性细胞RS-H526,并向H526以及RS-526细胞中转染si-ABCG2、pc-DNA3.1-ABCG2及其阴性对照或者转染miR-374bmimics、inhibitors,共转染mimics+si-ABCG2、inhibitors+si-ABCG2。采用qRT-PCR检测ABCG2mRNA表达水平,MTT检测细胞活力情况,划痕实验检测细胞迁移能力。结果 MTT结果显示,si-ABCG2组细胞在CPT-11诱导下的活力降低(t=5.02,P=0.047),并且迁移能力降低,差异有统计学意义,t=4.33,P=0.028。pc-ABCG2组细胞活力升高,t=2.44,P=0.003。miR-374b靶向抑制ABCG2表达,转染mimics的细胞在在CPT-11诱导下的活力(t=6.54,P=0.004)和迁移能力降低(t=10.58,P=0.005),共转染si-ABCG2进一步抑制细胞活力(t=7.52,P=0.019)和迁移能力(t=9.58,P=0.008);转染inhibitors提高非耐药肺癌细胞在CPT-11诱导下的活力(t=4.628,P=0.022),但是被共转染si-ABCG2抑制,t=7.519,P=0.003。结论 miR-374b靶向抑制ABCG2表达,激活CPT-11耐药性SCLC细胞对药物化疗的敏感性,可抑制肿瘤细胞活力、促进肿瘤细胞凋亡。OBJECTIVE ABCG2 was reported to promote cell drug-resistance,whose over-expression was closely related to cell drug-resistance occurring.According to previous investigation,miR-374 b,a common tumor suppressor gene could inhibit tumor cell viability and prompt apoptosis.In this study,we aimed to investigate the targeting relationship between miR-374 band ABCG2 and the effects in drug-resistance forming of small cell lung cancer.METHODS Twenty SCLC tissues and 20 paracancer tissues,12 male cases and 8 female cases,in the First Affiliated Hospital of Hebei North University were chosen for investigation.All experimental data are mean of three parallel experiments.Irinotecan was employed to construct resistance cells RS-H526 and H526 were transfected with si-ABCG2,pc-DNA3.1-ABCG2 miR-374 b mimics,mimics+si-ABCG2,inhibitors+si-ABCG2.qRT-PCR was utilized to detect ABCG2 mRNA expression level,MTT assay was performed for cell viability,and wound healing analysis was for exploring cell migration ability.RESULTS MTT assay results showed that the migration(t=5.02,P=0.047)and viability(t=5.02,P=0.047)of cancer cells in si-ABCG2 group were suppressed when treated with Irinotecan,but that of cells in pc-ABCG2 reduced(t=2.44,P=0.003)miR-374 binhibited ABCG2 expression;the migration(t=10.58,P=0.005)and viability(t=6.54,P=0.004)of cancer cells in mimics group was suppressed when treated with Irinotecan.When cells treated with Irinotecan were co-transfected with si-ABCG2 and mimics,their migration(t=7.52,P=0.019)and viability(t=9.58,P=0.008)were further hindered.Inhibitor transfection significantly promoted normal lung cancer cells viability when induced by Irinotecan,which was inhibited by co-transfecting si-ABCG2(t=7.519,P=0.003).CONCLUSIONS MiR-374 breverses the drug-resistance of small cell lung cancer to Irinotecan by targeting ABCG2 expression,which can inhibit the activity of tumor cells and promote the apoptosis of tumor cells.

关 键 词：miR-374b ABCG2 伊立替康 增殖 迁移 

分 类 号：R734.2[医药卫生—肿瘤]