胃癌SGC7901细胞长春新碱耐药性相关miRNA的初步分析  

Preliminary analysis of miRNA expression profile of SGC7901/VCR cell lines

作　　者：陶一明[1] 陈子华[2] 陈晋湘[2] 陈志康[2] 葛杰[2] 

机构地区：[1]中南大学湘雅医院医学科学研究中心,湖南长沙410008 [2]中南大学湘雅医院普通外科,湖南长沙410008

出　　处：《中国现代医学杂志》2013年第18期45-50,共6页China Journal of Modern Medicine

摘　　要：目的探讨胃癌SGC7901细胞对长春新碱(VCR)化疗药物耐药相关的微小RNA(miRNA),并预测异常表达miRNA的靶基因。方法体外培养胃癌SGC7901细胞和胃癌VCR耐药细胞SGC7901/VCR;提取总RNA并质检,采用miRNA 8.0微阵列芯片检测分析miRNA表达谱;荧光定量RT-PCR(qRT-PCR)验证差异表达的miRNA,生物信息学分析软件预测可能调控的靶基因;采用Western blot方法检测靶基因的蛋白表达水平。结果芯片检测发现,SGC7901和SGC7901/VCR细胞中63个异常表达的miRNA;其中miR-1267、miR-221、miR-223、miR-200c、miR-99a表达显著性上调;miR-122、miR-1246、miR-302a、miR-195、miR-124表达显著性下调。在胃癌细胞和胃癌组织中验证结果初步显示,miRNA-122和miR-302a可能分别调控靶基因IGF-IR和WDR62。结论获得的差异表达miRNA,以及预测的靶基因IGF-IR和WDR62可能在胃癌化疗药物VCR耐药中起关键性作用。To investigate the expression profile of specific microRNA（miRNA） of SGC7901/VCR cell lines,and potential miRNA target genes were analyzed by bioinformatics.【Methods】SGC7901 and SGC7901/ VCR cell lines were cultured,the total RNA was isolated and examined.The expression profiles of miRNA were detected with miRNA 8.0 type microarray chip and the compared array results were confirmed by real time PCR.Potential miRNA targets were analyzed.The microarray results were validated by Real-Time PCR and Western blot in gastric cancer cell lines and tissue.【Results】miRNA microarray detection showed that there were 63 miRNA with different expression in SGC7901 and SGC7901/VCR cell,among which miR-1267,miR-221,miR-223,miR-200c and miR-99a were significantly up-regulated,and miR-122,miR-1246,miR-302a,miR-195,miR-124 were significantly down-regulated.Based on the expression of miRNA-122 and miRNA-1246 in gastric cancer cell lines and tissues,both IGF-IR and WDR62 may be important target genes.【Conclusions】miRNA-122 and miR-302a may be involved in the multidrug resistance of gastric cancer by regulating their target genes IGF-IR and WDR62.

关 键 词：胃癌 MIRNA SGC7901 VCR 多药耐药 

分 类 号：R655.4[医药卫生—外科学]