Hepatobiliary and Intestinal Surgery Research Center, Central South University, Changsha 410008, China;
Department of Liver Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China.
Hepatobiliary Surg Nutr. 2013 Apr;2(2):78-83. doi: 10.3978/j.issn.2304-3881.2012.12.12.
To investigate the effects of EZH2 gene on the growth and migration of hepatocellular carcinoma HepG2 cell in vitro and in vivo.
EZH2 shRNA plasmid vectors were constructed and transfected into HepG2 cells. A model of EZH2 gene-silencing HepG2 cell lines was constructed, and the experimental cells were classified into 3 groups: HepG2 blank control group, HepG2-V vector control group and HepG2-EZH2 (-) group. The mRNA and protein expressions of EZH2 in these three cells were detected by real-time fluorogenic quantitative PCR and Western blotting, respectively. Cell proliferation was analyzed by MTT assay. Cells were inoculated subcutaneously in nude mice, and the growth of tumor cells in vivo was observed. Transwell chamber assay was performed to observe any change in the migration ability of cells.
The mRNA expression of HepG2 and HepG2-V was 100% and (95.27±10.87)%, respectively. Compared with the control group, the mRNA level of HepG2-EZH2 (-) were significantly decreased to (20.55±13.21)% (P<0.001). Similarly, the EZH2 protein expression were inhibited in HepG2-EZH2 (-) cells. The inhibition rate of tumor growth was 36.3% in vitro and 52.5% in vivo. The migration rate of the HepG2-EZH2 (-) group [(7.15±1.13)%] was significantly lower than those in the HepG2 group [(14.57±4.32)%] and the HepG2-V group [(15.21±5.22)%], with significant differences (both P<0.05).
EZH2 silencing can effectively inhibit the proliferation and growth of HepG2 cells in vitro and in vivo and inhibit cell migration. Therefore, the EZH2 gene may be a novel target for the treatment of liver cancer.
研究 EZH2 基因对体外和体内肝癌 HepG2 细胞生长和迁移的影响。
构建 EZH2 shRNA 质粒载体并转染 HepG2 细胞。构建 EZH2 基因沉默 HepG2 细胞系模型，将实验细胞分为 3 组：HepG2 空白对照组、HepG2-V 载体对照组和 HepG2-EZH2（-）组。采用实时荧光定量 PCR 和 Western blot 分别检测这 3 组细胞中 EZH2 的 mRNA 和蛋白表达。MTT 法分析细胞增殖。将细胞接种于裸鼠皮下，观察体内肿瘤细胞生长情况。Transwell 小室法观察细胞迁移能力变化。
HepG2 和 HepG2-V 的 mRNA 表达率分别为 100%和（95.27±10.87)%。与对照组相比，HepG2-EZH2（-）组的 mRNA 水平显著降低至（20.55±13.21)%（P<0.001）。同样，HepG2-EZH2（-）细胞中 EZH2 蛋白表达受到抑制。体外抑制肿瘤生长率为 36.3%，体内抑制肿瘤生长率为 52.5%。HepG2-EZH2（-）组的迁移率[（7.15±1.13)%]明显低于 HepG2 组[（14.57±4.32)%]和 HepG2-V 组[（15.21±5.22)%]，差异均有统计学意义（均 P<0.05）。
EZH2 沉默可有效抑制 HepG2 细胞在体外和体内的增殖和生长，并抑制细胞迁移。因此，EZH2 基因可能成为肝癌治疗的新靶点。
