Li Jianfang, Ding Shixiong, Ying Liping, Hu Airong, Hu Yaoren, Liang Xiaoyue
The Second People's Hospital of Ningbo, Ningbo 315010, China.
Zhonghua Gan Zang Bing Za Zhi. 2014 May;22(5):354-7. doi: 10.3760/cma.j.issn.1007-3418.2014.05.007.
To explore the effect of silencing the Notch2 gene by small interfering (si)RNA on the proliferation of the HepG2 human hepatocellular carcinoma (HCC) cells.
Notch2-siRNA was transfected as a liposomal formulation into HepG2 cells. 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 RTPCR, 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) colorimetric assay.
The untransfected HepG2 cells showed significantly upregulated transcript expression of Notch2, and not of Notch1, Notch3 or Notch4, compared to the other non-HCC cell lines. Following transfection of Noteh2-siRNA into HepG2 cells, the mRNA expression of Notch2 and Hes1 and the protein expression of Notch2 were significantly decreased. The rales of proliferation inhibition in HepG2 following transfection of Notch2-siRNA showed an increasing time-related trend, with 2.64% ± 1620% 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 of transfection was significantly different from that of the groups of control cells.
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.
探讨小干扰(si)RNA沉默Notch2基因对人肝癌HepG2细胞增殖的影响。
将Notch2-siRNA以脂质体制剂形式转染至HepG2细胞。非肝癌细胞系SG07901(胃癌)和SW620(结肠癌)用作对照。通过RT-PCR检测Notch2和Hes1的mRNA表达,通过蛋白质印迹法检测Notch2的蛋白质表达。采用细胞计数试剂盒-8(CCK8)比色法评估转染后HepG2细胞的增殖情况。
与其他非肝癌细胞系相比,未转染的HepG2细胞中Notch2的转录本表达显著上调,而Notch1、Notch3或Notch4则未上调。将Notch2-siRNA转染至HepG2细胞后,Notch2和Hes1的mRNA表达以及Notch2的蛋白质表达均显著降低。转染Notch2-siRNA后HepG2细胞的增殖抑制率呈时间相关的增加趋势,12小时时为2.64%±1.620%,24小时时为38.34%±8.80%,48小时时为70.05%±7.80%,72小时时为70.78%±10.00%,96小时时为74.22%±4.80%。转染24小时时的抑制率与对照细胞组有显著差异。
在常见的肝癌培养细胞系HepG2中Notch2上调。siRNA介导的Notch2沉默对HepG2增殖具有抑制作用,提示该方法作为肝癌靶向治疗的潜力。