尾型同源盒基因2沉默对人胃癌耐顺铂细胞SGC7901／DDP多药耐药性的逆转  

作　　者：罗文[1] 杨杰[1] 廉超[1] 王晓通[1] 谢玉波[2] 肖强[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,南宁530021 [2]广西医科大学第一附属医院麻醉科,南宁530021

出　　处：《中华消化外科杂志》2014年第1期55-61,共7页Chinese Journal of Digestive Surgery

基　　金：国家自然科学基金（81060201）;广西自然科学基金（2011GXNSFA018273）;重大疾病防治新技术研究开发（桂科攻1298003-2-6）

摘　　要：目的探讨尾型同源盒基因2（Cdx2）沉默对逆转人胃癌耐顺铂细胞SGC7901／DDP多药耐药性的影响。方法将对数生长期的人胃癌耐顺铂细胞SGC7901／DDP接种于培养板中，分为3组：感染RNA干扰Cdx2基因沉默重组慢病毒载体（pLL—Cdx2-shRNA）作为实验组；感染慢病毒空载体作为阴性对照组；空白对照组不做任何处理。Westernblot及RT—PCR检测Cdx2及凋亡相关基因C—myc、cyclinD1、survivin等蛋白及mRNA表达水平；MTT法检测3组细胞对化疗药物阿霉素、5．氟尿嘧啶、顺铂的敏感性；流式细胞技术检测3组细胞阿霉素的泵出率、细胞周期分布及细胞凋亡情况。计量资料用面±s表示，多组间比较采用单因素方差分析，两组间比较采用SNK—q检验，计数资料采用，检验。结果实验组、阴性对照组、空白对照组人胃癌耐顺铂细胞SGC7901／DDP中各蛋白相对表达量：Cdx2分别为0．187±0．060、0．535±0．033、0．567±0．014，C—myc分别为0．086±0．004、0．379±0．006、0．354±0．004，cyclin D1分别为0．016±±0．005、0．141±0．003、0．162±0．008，survivin分别为0．276±0．012、0．672±0．009、0．517±0．313，与阴性对照组和空白对照组比较，实验组上述蛋白表达量明显下降，差异有统计学意义（F=247．385，3．353，597．882，98．628，P〈0．05）。实验组、阴性对照组、空白对照组mRNA相对表达量：cdx2分别为0．184±0．010、0．894±0．056、0．837±0．049，c—myc分别为0．212±0．022、0．538±0．021、0．545±0．032，cyclinD1分别为0．045±0．009、0．163±0．009、0．157±0．010，survivin分另4为0．401±0．027、0．824±0．016、0．782±0．056，与阴性对照组和空白对照组比较，实验组上述基因mRNA表达量明显下降，差异有统计学意义（F=243．776，161．793，138．523，118．426，P〈0．05）。MTT法检测实验组、阴性对照组和空白对照组人胃癌Objective To investigate the effects of caudal type homeobox 2 (Cdx2) silence on reservion of multi-drug resistance of gastric carcinoma cisplantin-resistant cell SGC7901/DDP.Methods Gastric carcinoma cisplantin-resistant cells SCG7901/DDP in the logarithmic phase were cultured in the plate,and were divided into the experimental group [gastric carcinoma cells of SGC7901/DDP were infected with a silent Cdx2-recombinanted lentiviral vector (pLL-Cdx2-shRNA)],the negative control group (gastric carcinoma cells of SGC7901/DDP were infected with empty lentiviral vector) and the blank control group (gastric carcinoma cells of SGC7901/DDP were not treated).The protein and mRNA expressions of Cdx2 and apoptosis related genes like c-myc,cyclin D1 and survivin were detected by the Western blot and reverse-transcription PCR,respectively.The sensitivity of the cells in the 3 groups to adriamycin,5-fluorouracil and cisplatium were assessed by MTT.The pump-out rate of adriamycin,cell cycle distribution and apoptosis of the 3 groups were analyzed using flow cytometry.All measurement data were expressed with mean ± standard deviation.Comparison among multi-groups was done by one-way analysis of variance,and comparison between 2 groups was done by SNK-q test.The enumeration data were analyzed using the chi-square test.Results The relative protein expression levels of Cdx2,c-myc,cyclin D1 and survivin were 0.187 ± 0.060,0.086 ± 0.004,0.016 ± 0.005 and 0.276 ± 0.012 in the experimental group,0.535 ± 0.033,0.379 ± 0.006,0.141 ± 0.003 and 0.672 ± 0.009 in the negative control group,and 0.567 ± 0.014,0.354 ± 0.004,0.162 ± 0.008 and 0.517 ± 0.313 in the blank control group,respectively.The relative protein expression levels of Cdx2,c-myc,cyclin D1 and survivin in the experimental group were significantly lower than those in the negative control group and the blank control group (F =247.385,3.353,597.882,98.628,P ＜0.05).The relative mRNA expression levels of Cdx2,c-myc,cyclin D1 and survivin were 0.184 ± 0.010,0.212 ± 0.

关 键 词：胃肿瘤 尾型同源盒基因-2 胃癌耐顺铂SGC7901细胞 多药耐药性 

分 类 号：R735.2[医药卫生—肿瘤]