摘要
目的研究RhoC基因沉默对肝癌细胞生长和转移潜力的影响。方法基因重组法构建小干扰RNA表达载体,沉默肝癌细胞RhoC基因表达;平板克隆实验检测细胞生长潜力,半定量RT-PCR法检测细胞周期蛋白和MMPs表达水平。结果RNAi组克隆形成百分数显著低于对照组(20.33±5.42%和70.58±10.10%,P<0.01);RNAi组细胞CyclinD1表达显著低于对照组(0.45±0.21和1.25±0.24,P<0.05),而P21基因表达则显著高于对照组(1.07±0.23和0.47±0.11,P<0.05);RNAi组MMP-2基因表达显著低于对照组(0.38±0.13和1.41±0.25,P<0.01);与MMP-2相似,RNAi组MMP-9基因表达水平也显著低于对照组(0.44±0.17和1.36±0.18,P<0.01)。结论基因沉默RhoC能够显著抑制肝癌细胞生长和转移潜力,为靶向RhoC肿瘤基因治疗奠定了实验基础。
Objective To study tile effects of RhoC gene silencing on the potentiality of hepatocellular carcinoma cell （HCC） gro＇,wth and metastasis. Methods Clone the syntiresized oligoneucleotide fragment into RNAJ plasmid and transfect it into HCC to silence RhoC expression; Assay the proliferation potential of cells by plate cloning test and determine the mRNA levels of CyclinD1 and P21 by semi-quantitative RT-PCR; Assay tumor metastasis potentiality through determining MMP-2 and MMP-9 mRNA levels by semi-quantitative RT-PCR. Results The clone formation rate in RNAi group was significandy lower than control（20.33±5.42% and 70.58 ±10.10 %, P 〈 0.01）. CyclinD1 mRNA level in RNAi group was notably lower than control（ 0.45±0.21 and 1.25±0.24, P 〈 0.05）, while P21 mRNA level was significantly higher than control（ 1. 07 ± 0.23 and 0.47 ± 0.11, P 〈 0.05）. MMP-2 mRNA level in RNAi was markedly lower than control（0.38 ± 0.13 and 1.41 ± 0.25, P 〈 0.01）.Similar to MMP-2, MMP-9 mRNA level in RNAi group was also significantly lower than control（0.44 ±0.17 and 1.36±0.18, P 〈 0.01） .Conclusion RhoC gene silencing can inhibit the potentiality of HCC proliferation and metastasis,laying a foundation to tumor gene therapy against RhoC.
出处
《中国实验诊断学》
2008年第10期1212-1215,共4页
Chinese Journal of Laboratory Diagnosis
