小干扰RNA下调Notch2对肝细胞癌HepG2细胞增殖的影响  

Effect of siRNA-mediated silencing of Notch2 on proliferation of the HepG2 human hepatocellular carcinoma cells

作　　者：李建芳[1] 丁世雄 应丽萍 胡爱荣 胡耀仁 梁晓岳 

机构地区：[1]宁波市第二医院肝二科,315010 [2]宁波市肿瘤分子生物学重点实验室 [3]宁波市肝病研究所

出　　处：《中华肝脏病杂志》2014年第5期354-357,共4页Chinese Journal of Hepatology

摘　　要：目的研究小干扰RNA（siRNA）干扰Notch2对HepG2细胞增殖的影响。方法用siRNA干扰HepG2细胞中Notch2基因，用RT-PCR检测siRNA干扰后Notch2mRNA与下游靶基因HeslmRNA的表达水平，用Westernblot方法检测Notch2蛋白的改变情况，然后用CCK8法测定HepG2细胞增殖情况，用析因设计的方差分析和组间t检验进行统计学比较。结果肝细胞癌HepG2细胞中Notch2mRNA高表达，Notch2-siRNA干扰抑制后，HepG2细胞的Notch2mRNA和蛋白水平下调，下游Hesl的mRNA表达水平降低。HepG2细胞在Notch2-s识NA处理组12、24、48、72h和96h的增殖抑制率分别为2．640／0±16．20％、38．340／0±8．8%、70．05％±7．80％、70．78％±10．00％和74．22％±4．80％，对照组siRNA处理的细胞增殖抑制率分别为-0．21％±3．65％、11．61％±10．66％、22．11％±7．93％、7．64％±10．00％和4．18％±7．64％，两组比较，F=5．48，P〈0．01，差异有统计学意义。与对照组相比，24h后增殖差异均有统计学意义（t值分别为3．405、7．473、7．749和13．460，P值均〈0．05）。结论Notch2在体外培养的肝细胞癌HepG2细胞中高表达，下调Notch2可以减缓HepG2细胞的增殖，为Notch2的靶向干预作为肝细胞癌的治疗提供了有价值的实验依据。Objective To explore the effect of silencing the Notch2 gene by small interfering （si）RNA on the proliferation of the HepG2 human hepatoceUular carcinoma （HCC） cells. Methods Notch2-siRNA was transfeeted as a liposomal formulation into HepG2 ceils. The non-HCC cell lines SG07901 （gastric cancer） and SW620 （colon cancer） were used as controls. The mRNA expression of Notch2 and Hesl were detected by RT- PCR, and the protein expression of Notch2 was detected by western blotting. The proliferation of transfected HepG2 cells was assessed by the cell counting kit-8 （CCK8） colofimetric assay. Results The untransfected HepG2 cells showed significantly upregulated Wanscript expression of Notch2, and not ofNotchl, Notch3 or Notch4, compared to the other non-HCC cell lines. Following transfection of Notch2-siRNA into HepG2 cells, the mRNA expression of Notch2 mad Hesl and the protein expression of Notch2 were significantly decreased. The rates of proliferation inhlbifion in HepG2 following iransfection of Notch2-siRNA showed an increasing time-related trend, with 2.64% ± 16.20% at 12 h, 38.34% ± 8.80% at 24 h, 70.05%± 7.80% at 48 h, 70.78% ± 10.00% at 72 h, and 74.22% ±4.80% at 96 h. The inhibition rate at 24 h oflransfection was significantly different from that of the groups of control cells. Conclusion Notch2 is upregulated in the common HCC cultured cell line HepG2. siRNA-mediated silencing of Notch2 exerts inhibition effects on HepG2 proliferation, suggesting the potential for this approach as targeted therapy for treating HCC.

关 键 词：癌 肝细胞 细胞增殖 受体 NOTCH 

分 类 号：R735.7[医药卫生—肿瘤]