miR-802对叉头框转录因子M1抑制A549/DDP细胞顺铂耐药性的靶向调控作用  

Targeting regulation of miR-802 on drug-resistance of A549/DDP cells to cisplatin through downregaluting forkhead box protein M1

作　　者：李志伟[1] 尚勇[1] 郭庆伟[1] LI Zhiwei;SHANG Yong;GUO Qingwei(Department of Cardiothoracic Surgery,Zhoukou Central Hospital.Zhoukou 466000,China)

机构地区：[1]周口市中心医院胸心外科,河南周口466000

出　　处：《中华实用诊断与治疗杂志》2020年第2期117-120,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河南省2016年科技发展计划(162102310215)。

摘　　要：目的探讨miR-802抑制非小细胞肺癌A549/DDP细胞的顺铂耐药性及其对叉头框转录因子M1(forkhead box protein M1, FoxM1)的靶向调控作用。方法 A549、A549/DDP细胞采用反转录PCR法检测miR-802相对表达量。将A549/DDP细胞分为空白转染组和miR-802过表达组(过表达组),分别转染miR-NC和miR-802类似物(miR-802 mimics),转染24、48、72、96 h后,采用反转录PCR法检测2组细胞miR-802相对表达量,并采用CCK-8法检测2组细胞增殖率;转染48 h后,CCK-8法检测转染后A549/DDP细胞对DDP的敏感性,采用流式细胞仪检测2组细胞凋亡率和细胞周期,采用Western blot法检测转染后A549/DDP细胞内FoxM1蛋白相对表达量。结果 A549/DDP细胞miR-802相对表达量(0.21±0.03)低于A549细胞(0.85±0.12)(P<0.05);转染24、48、72、96 h,过表达组细胞miR-802相对表达量依次增高(P<0.05),且均高于空白转染组(P<0.05);转染48、72、96 h,空白转染组细胞增殖率依次增高(P<0.05),且均高于过表达组(P<0.05);转染48 h,过表达组细胞半数抑制浓度[(35.28±2.17)mg/L]和细胞早期凋亡率[(17.2±1.1)%]均高于空白转染组[(14.22±1.28)mg/L、(9.0±0.8)%](P<0.05),S期和G2/M期细胞比率[(21.30±0.20)%、(8.35±0.33)%]及细胞内FoxM1蛋白相对表达量(0.21±0.04)均低于空白转染组[(27.54±0.52)%、(14.67±0.70)%、0.44±0.06](P<0.05)。结论 miR-802可能通过抑制FoxM1表达而降低非小细胞肺癌A549/DDP细胞的顺铂耐药性。Objective To investigate the effect of miR-802 on suppressing the drug-resistance of non-small cell lung cancer A549/DDP cells to cisplatin and its targeting regulation of forkhead box protein M1(FoxM1). Methods Reverse transcription PCR was used to detect the expression of miR-802 in A549 and A549/DDP cells. The A549/DDP cells were divided into blank transfection group and miR-802 overexpression group, transfected with miR-NC and miR-802 mimics, respectively. After 24, 48, 72 and 96 h of transfection, reverse transcription PCR was used to detect the relative expression of miR-802, and CCK-8 method was used to detect the proliferation of cells. After 48 h of transfection, the sensitivity of A549/DDP cells to DDP was detected by CCK-8 method, the apoptosis rate and cell cycle were detected by flow cytometry, and the relative expression of FoxM1 protein in A549/DDP cells was detected by Western blot. Results The relative expression of miR-802 was lower in A549/DDP cells(0.21±0.03) than that in A549 cells(0.85±0.12)(P<0.05). After 24, 48 72 and 96 h of transfection, the relative expression of miR-802 increased gradually in miR-802 overexpression group(P<0.05), all of which were higher than those in blank transfection group(P<0.05). After 48 h of transfection, the half inhibitory concentration of cells and the early cell apoptosis rate were higher in miR-802 overexpression group((35.28±2.17) mg/L,(17.2±1.1)%) than those in blank transfection group((14.22±1.28) mg/L,(9.0±0.8)%)(P<0.05), while the ratios of S-phase and G2/M-phase cells((21.30±0.20)%,(8.35±0.33)%) and the relative expression of FoxM1 protein(0.21±0.04) were lower than those in blank transfection group((27.54±0.52)%,(14.67±0.70)%, 0.44±0.06)(P<0.05). Conclusion miR-802 may reduce the drug resistance of A549/DDP cells to cisplatin by suppressing the expression of FoxM1.

关 键 词：非小细胞肺癌 miR-802 叉头框转录因子M1 A549/DDP细胞 顺铂 耐药性 

分 类 号：R734.2[医药卫生—肿瘤]