Xiao Zhen-yu, Chen Xiao-ping, Huang Zhi-yong
Center of Hepatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Gan Zang Bing Za Zhi. 2005 Oct;13(10):768-71.
To investigate the effect of antisense cDNA of cyclin D1 on the cyclin D1 gene expression and cell proliferation of human hepatocarcinoma HepG2 cells in vitro.
Plasmids containing cyclin D1 antisense cDNA were constructed and transfected into HepG2 cells. Their effects on cell proliferation were examined by MTT method, RT-PCR, immunohistochemical means, and flow cytometry.
Cyclin D1 antisense cDNA significantly inhibited the growth of HepG2 cells. The inhibition peaked at 48 hour after transfection by MTT method. RT-PCR analysis showed that cyclin D1 antisense cDNA down-regulated cyclin D1 at the mRNA levels. Expression level of cyclin D1 protein was also decreased as shown by immunohistochemical studies. Cell-cycle analysis by flow cytometry showed that transfected HepG2 cells were arrested at the G1 phase of the cell cycle.
Our data suggest that cyclin D1 antisense cDNA could specifically inhibit the expression of cyclin D1 mRNA and protein and regulate cell cycle and cell proliferation of HepG2 cells. Cyclin D1 antisense cDNA may serve as a potential antitumor strategy in regulating cell-cyclin treating advanced HCCs.
研究细胞周期蛋白D1反义cDNA对人肝癌HepG2细胞体外细胞周期蛋白D1基因表达及细胞增殖的影响。
构建含细胞周期蛋白D1反义cDNA的质粒并转染至HepG2细胞。通过MTT法、RT-PCR、免疫组织化学方法及流式细胞术检测其对细胞增殖的影响。
细胞周期蛋白D1反义cDNA显著抑制HepG2细胞生长。MTT法检测显示转染后48小时抑制作用达到峰值。RT-PCR分析表明细胞周期蛋白D1反义cDNA在mRNA水平下调细胞周期蛋白D1。免疫组织化学研究显示细胞周期蛋白D1蛋白表达水平也降低。流式细胞术进行的细胞周期分析表明，转染的HepG2细胞停滞于细胞周期的G1期。
我们的数据表明，细胞周期蛋白D1反义cDNA可特异性抑制细胞周期蛋白D1 mRNA和蛋白的表达，并调节HepG2细胞的细胞周期和细胞增殖。细胞周期蛋白D1反义cDNA可能作为一种潜在的抗肿瘤策略用于调控细胞周期治疗晚期肝癌。
