沉默c-met基因表达对胃癌肝高转移潜能细胞生物学行为的影响  

Effects of Silencing of c- met Gene Expression on Biological Behavior of Gastric Cancer Cells With High Livermetastatic Potential

作　　者：沈皓[1] 呼圣娟[2] 师红利[2] 姜荣兴[3] 贺婵婵 张蓉[2] 

机构地区：[1]宁夏银川市第一人民医院消化内科,750001 [2]宁夏回族自治区人民医院消化内科 [3]宁夏银川市第一人民医院放射科,750001

出　　处：《中国全科医学》2016年第3期296-299,共4页Chinese General Practice

基　　金：国家自然科学基金资助项目(81060193);宁夏自然科学基金资助项目(NZ1283)

摘　　要：背景胃癌肝高转移潜能细胞(XGC9811-L)是本实验组在前期工作中建立的一株具有肝脏高转移潜能的胃癌细胞系,c-met基因在其中高表达。c-met是肝细胞生长因子(HGF)的受体。目的研究沉默c-met基因表达对XGC9811-L的增殖、侵袭和转移等生物学行为的影响。方法 2014年1—6月,采用细胞转染法将重组质粒p SuppressorRetro-c-met-siRNA导入XGC9811-L,建立稳定转染细胞系,按有无转染和转染目的基因的不同分为未转染组、空载体组、随机序列组和siRNA组。采用荧光定量反转录-聚合酶链式反应(RT-PCR)法检测XGC9811-L中c-met mRNA相对表达水平,Western blotting法检测c-met相对表达水平,通过细胞增殖实验检测各组细胞增殖能力并绘制细胞生长曲线,通过侵袭实验和运动实验计算各组穿膜细胞数。结果 siRNA组c-met mRNA、c-met相对表达水平低于未转染组、空载体组、随机序列组(P<0.05)。siRNA组第3天、第5天、第6天、第7天细胞增殖能力低于未转染组、空载体组、随机序列组(P<0.05)。在侵袭实验中,siRNA组穿膜细胞数少于未转染组、空载体组、随机序列组(P<0.05)。在运动实验中,siRNA组穿膜细胞数少于未转染组、空载体组、随机序列组(P<0.05)。结论沉默c-met基因的表达可以抑制XGC9811-L的增殖、侵袭和转移能力,c-met在胃癌肝转移的发生、发展过程中可能起重要作用,抑制c-met基因表达可能会抑制胃癌细胞向肝脏转移。Background XGC9811 - L, which is a gastric cancer cell line with high liver - metastatic potential, was established in the preliminary work of the study. The expression of c - met is higher in XGC9811 - L than in other gastric cancer cell lines, and c - met is the receptor of hepatocyte growth factor. Objective To research the effects of silencing of c - met gene expression on biological behaviors of XGC9811 - L, such as proliferation, invasion and metastasis. Methods From January to June 2014, using cell transfection method, we imported recombinant plasmid pSuppressorRetro -c -met -siRNA into XGC9811 -L cells and built stable transfection cell lines. Based on whether transfection was made and the difference in the target genes of transfeetion, the stable transfection cell lines were divided into four groups： non - transfection group, empty carrier group, random sequence group and siRNA group. Fluorescent quantitation RT-PCR method was adopted to detect the relative expression level of c - met mRNA in XGC9811 - L; c - met relative expression was measured by western blotting method. By cell proliferation assay, the cell proliferation ability of each group was detected, and cell growth curves were drawn. Invasion assay and movement experiment were conducted to calculate the number of transmembrane cells of each group. Results siRNA group was lower than the other three groups in the relative expression levels of c - met mRNA and c - met （ P 〈 0. 05 ） . siRNA group was lower than the other three groups in cell proliferation ability on day 3, day 5, day 6 and day 7 （ P 〈 0. 05 ） . In invasion assay, siRNA group was less than the other three groups in the number of transmembrane cells （ P 〈 0. 05 ） . In movement experiment, siRNA group was less than other three groups in the number of transmembrane cells （ P 〈 0. 05 ） . Conclusion Silencing of c - met gene expression can inhibit the proliferation, invasion and metastasis of XGC9811 - L, and c - met may play an important part in the occurrence

关 键 词：RNA 小分子干扰 基因沉默 原癌基因蛋白质C-MET XGC9811-L 

分 类 号：R735.2[医药卫生—肿瘤]