血红素加氧酶-1小干扰RNA对人胃癌9811-P细胞体外侵袭的影响  

Effects of Heme Oxygenase-1 small interfering RNA on invasion of human gastric cancer 9811-P cells in vitro

作　　者：祁妮娜[1] 白飞虎[2] 

机构地区：[1]宁夏医科大学研究生院,宁夏银川750004 [2]宁夏医科大学附属医院消化内科,宁夏银川750004

出　　处：《中国现代医学杂志》2011年第17期1953-1957,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金资助项目(No:30860332)

摘　　要：目的构建血红素加氧酶-1(HO-1)小干扰RNA(siRNA)表达载体,观察HO-1siRNA对人胃癌9811-P(GC9811-P)细胞体外侵袭转移能力的影响。方法设计HO-1siRNA序列,构建其真核表达载体,在Lipofectamine2000介导下转染GC9811-P细胞,通过Western-blotting及RT-PCR检测HO-1在蛋白及mRNA水平的表达变化。通过细胞划痕实验、Transwell小室实验观察GC9811-P细胞体外侵袭转移能力。结果成功构建了HO-1siRNA表达载体,并命名为pEGFP-C1/HO-1-siRNA。获得成功转染HO-1-siRNA的GC9811-P细胞,mRNA及蛋白水平均显示HO-1在GC9811-P细胞中明显下降。细胞划痕实验结果显示转染了HO-1-siRNA组的GC9811-P细胞迁移不明显,而转染了阴性对照组细胞和未转染的空白对照组细胞大量迁移至划痕区。Transwell小室实验结果显示转染了HO-1-siRNA组的GC9811-P细胞穿膜数为(54.2±2.68),而转染了阴性对照组的细胞穿膜数为(83.2±5.71),未转染的空白对照组细胞穿膜数为(84.3±4.29),LSD-t检验分析各组间细胞穿膜数的差异,HO-1-siRNA组细胞穿膜数小于阴性对照组及空白对照组细胞穿膜数(P<0.05)。结论 HO-1在GC9811-P细胞体外侵袭转移中发挥着重要作用,HO-1-siRNA可抑制GC9811-P细胞体外侵袭转移能力。[Objective] To construct HO-1 small interfering RNA expression vector, observe the effects in invasion and metastasis of HO-1 siRNA on gastric cancer 9811-P cells in vitro. [Methods] Design HO-1 small interfering RNA sequence, construct its eukaryotic expression vector, in the lipofectamine 2000-mediated transfection of GC9811-P cells. Detect the HO-1 expression in protein and mRNA levels by western-blot and RT-PCR. Through wound-healing assay and transwell assay to observe the invasion and metastasis of GC9811-P cells in vitro. [Results] Construction of HO-1 small interfering RNA expression vector pEGFP-C1/ HO-I-siRNA was successfully. Getting successful transfeetion of the HO-I-siRNA of GC9811-P ceils, mRNA and protein levels show that HO-1 expression was decreased significantly in GC9811-P cells. Wound-healing assay show that transfect with HO-I-siRNA group GC9811-P cells migration is not obvious, but a large number of negative control group transfect cens and untransfect control cells moved to the scratch area. Transwell assay show that transfect with HO-I-siRNA group GC9811-P cells in the number of penetrating are （54.2±2.68）, while the negative control group transfeet cells are （83.2±5.71）, untransfeet control group cells are （84.3±4.29）. LSD-t test analyzed the differences between groups of HO-I-siRNA cells are less than the number of negative control group and blank control group. [Conclusion] HO-1 play an important role in invasion and metastasis of GC9811-P cells in vitro, and the HO-I-siRNA may inhibit invasion and metastasis of GC9811-P ceils in vitro.

关 键 词：小干扰R.NA 胃癌细胞 侵袭 

分 类 号：R735.2[医药卫生—肿瘤] R979.1[医药卫生—临床医学]