RNA干扰GCF2基因沉默对人肝癌BEL-7404细胞侵袭能力的影响

发布时间：2018-04-30 07:32

  本文选题：GCF + 肝细胞癌　； 参考：《广东医学》2017年10期

【摘要】：目的探讨沉默GCF2基因表达对人肝癌BEL-7404细胞侵袭能力的影响。方法实验分为GCF2-siRNA组、NC-siRNA组及MOCK组,GCF2-siRNA组转染靶向GCF2基因的RNA干扰序列GCF2-siRNA,阴性对照组转染阴性对照序列NC-siRNA,MOCK组不转染任何序列。将靶向GCF2基因的RNA干扰序列(GCF2-siRNA)瞬时转染BEL-7404细胞以沉默GCF2表达,采用CCK-8法检测细胞增殖活性,体外侵袭实验检测细胞侵袭能力,Western blot检测基质金属蛋白酶(MMP)-9及GCF2靶基因MMP-23蛋白表达。结果转染GCF2-siRNA使GCF2表达下调,后者导致BEL-7404增殖受阻;GCF2-siRNA组的穿膜细胞数较阴性对照组(NC-siRNA)及未转染组(MOCK)明显减少(穿膜细胞数分别为41.5±0.95、61.3±1.57、64.5±1.65,P0.05);MMP-9和MMP-23蛋白表达较两个对照组明显下降(P0.05)。结论下调GCF2表达能抑制BEL-7404侵袭能力,提示GCF2促进肝癌细胞的侵袭可能是其参与肝癌发展进程的重要途径。
[Abstract]:Objective to investigate the effect of silent GCF2 gene expression on the invasiveness of human hepatocellular carcinoma BEL-7404 cells. Method experiments were divided into GCF2-siRNA group, NC-siRNA group and MOCK group, GCF2-siRNA group transfected RNA interference sequence of target GCF2 gene, negative control group transfected negative control sequence NC-siRNA, MOCK group did not transfect any sequence. The target GCF2 gene would be targeted. The RNA interference sequence (GCF2-siRNA) transiently transfected BEL-7404 cells to silence GCF2 expression, detected cell proliferation activity by CCK-8, and detected cell invasiveness in vitro, and Western blot detected the expression of MMP-23 protein of matrix metalloproteinase (MMP) -9 and GCF2 target gene. The number of membrane cells in the GCF2-siRNA group was significantly lower than that in the negative control group (NC-siRNA) and the untransfected group (MOCK) (the number of membrane cells were 41.5 + 0.95,61.3 + 1.57,64.5 + 1.65, P0.05), and the expression of MMP-9 and MMP-23 protein was significantly lower than that of the two control groups (P0.05). Conclusion the down-regulation GCF2 expression could inhibit BEL-7404 invasion ability, suggesting GCF2 promoted. Invasion of HCC cells may be an important way to participate in the development of HCC.

【作者单位】： 桂林医学院组胚教研室;桂林医学院临床医学院;
【基金】：广西高校科研项目(编号:KY2015LX284) 大学生创新创业训练计划项目(编号:201410601011,201510601003)
【分类号】：R735.7
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本文编号：1823648