Oct4B1基因过表达增强结直肠癌SW620细胞对奥沙利铂的耐药性及其机制  

Enhancing the oxaliplatin chemo - resistance via overexpression of Oct4B1 gene in SW620 colorectal cancer cells and its mechanism of drug resistance

作　　者：陈正权[1] 文坤明[1] 陈奕霖[1] 胡水清[1] 曾庆良[1] Chen Zhengquan;Wen Kunming;Chen Yilin;Hu Shuiqing;Zeng Qingliang(Department of C, astroenterological Surgery, the Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Chin)

机构地区：[1]遵义医学院附属医院胃肠外科,563003

出　　处：《中华实验外科杂志》2018年第5期858-861,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81260369）;贵州省科技厅科学技术基金[黔科合（J）字（2012）2365]

摘　　要：目的 探讨Oct4B1基因过表达是否增强结直肠癌SW620细胞对奥沙利铂耐药性及其机制,以及与诱导细胞上皮-间充质转化（EMT）过程的关系.方法 实验分组：将SW620细胞随机平均分为实验组及对照组、实验组（SW620-Oct4B1）：转染带G418抗性的Oct4B1过表达质粒;对照组（SW620-NC）：转染带G418抗性的阴性对照质粒;筛选合适浓度G418建立稳定转染细胞株.对稳定转染两组细胞作如下检测：（1）噻唑蓝（MTT）法检测细胞对不同浓度奥沙利铂的敏感性;（2）Western blot检测P-糖蛋白（P-gp）及EMT标志物E-钙黏蛋白（E-cadherin）、N-钙黏蛋白（N-cadherin）的表达;（3）划痕实验检测12、24 h迁移能力;（4）Transwell小室检测24 h时侵袭能力.结果 实验组与对照组比较：（1）对奥沙利铂敏感性实验组明显低于对照组（t =8.889、5.427、9.678、4.459、5.362,P=0.001、0.006、0.001、0.011、0.006）;（2）P-gp表达两组分别为0.401、0.439、0.419（0.420±0.020）及0.099、0.110、0.120（0.110 ±0.010）,实验组明显高于对照组（t=24.723,P=0.000）;（3）12 h伤痕愈合率分别为32.196％、32.382％、33.441％ （32.670 ±0.670）％及23.049％、25.286％、23.674％ （24.000±1.150）％,实验组明显高于对照组（f=11.245,P=0.001）;24 h伤痕愈合率分别为67.048％、68.650％、66.291％ （67.330±1.200）％和49.161％、47.632％、53.203％（50.000 ±2.880）％.实验组显著高于对照组（t=9.620,P=0.001）;（4）24 h两组透膜细胞数分别为400.105、407.500、384.300（397.300±11.850）个及205.641、186.244、215.02（202.300±14.680）个,实验组显著多于对照组（t=17.905,P=0.000）;（5）E-cadherin表达两组分别为0.186、0.190、0.223（0.200 ±0.020）及0.530、0.531、0.529（0.530 ±0.010）,实验组明显低于对照组（t=28.143,P=0.000）,而N-cadherin表达两组分别为0.949、0.935、0.906（0.930±0.020）及0.687、0.648、0.675（0.670±0.020）,实验�Objective To investigate whether enhancing the oxaliplatin chemo-resistance via over expression of Oct4B1 gene in SW620 colorectal cancer cells and its mechanism of drug resistance is associated with induction of cell epithelial mesenchymal transition （EMT）.Methods Experimental grouping.The SW620 cells were randomly divided into experimental and control groups,Group of Experiments：transfection with G418 resistance Oct4B1 expression plasmid was called the experimental group（SW620-Oct4B1）;negative control plasmid transfection with G418 resistance referred to as the control group （SW620-NC）.Screening suitable concentration of G418 to establish stable transfected cell line.The following detection of two groups of cells：（1） The sensitivity of the cells to the different concentrations of oxaliplatin was detected by methyl thiazol tetrazolium （MTT）;（2） Detecting the protein expression of P-glycoprotein （P-gp） and EMT marker （E-cadherin and N-cadherin）;（3）The migration ability of cell 12 h and 24 h was detected by scratch test;（4）Transwell cells were used for cell invasiveness when 24 h was detected.Resuits The experimental group was compared with the control group：（1） The sensitivity of the chemotherapy drug oxaliplatin sensitivity test group was significantly lower than that of the control group （t =8.889,5.427,9.678,4.459,5.362;P=0.001,0.006,0.001,0.011,0.006）;（2） The expression of Pgp in the experimental group and the control group was 0.401,0.439,0.419 （0.420 ± 0.020） and 0.099,0.110,0.120 （0.110 ±0.010）,and the experimental group was significantly higher than that of the control group （t =24.723,P =0.000）;（3） The healing rate of 12 h scar was 32.196%,32.382%,33.441% （32.670 ±0.670）% and 23.049%,25.286%,23.674% （24.000 ± 1.150）%,The experimental group was obviously higher than the control group （t =11.245,P =0.001）,and the healing rate of 24 h scar was 67.048%,68.650%,66.291% （67.330 ± 1.200）% and 49.161%,47.632%,53.203% �

关 键 词：结直肠癌 Oct4B1 上皮-间充质转化 耐药性 

分 类 号：R735.34[医药卫生—肿瘤]