论文部分内容阅读
目的在肝癌细胞系HepG2中稳定过表达HLrg,观察过表达的HLrg蛋白对HepG2的细胞周期和细胞形态等生物学影响。方法构建真核重组表达质粒pcDNA3.1(+)-hlrg,转染到HepG2细胞并进行稳定筛选;对稳定过表达pcDNA3.1(+)空载体的对照组、pcDNA3.1(+)-hlrg的实验组进行观察。Westernblot进行HLrg蛋白的鉴定。流式细胞仪测定细胞周期、MTT法测定生长曲线、透射电子显微镜观测细胞形态的改变。结果获得了稳定过表达pcDNA3.1(+)-hlrg的HepG2细胞株。与对照组相比,过表达HLrg蛋白的细胞株出现G1阻滞,微绒毛减少,分裂相减少,生长趋缓。结论稳定过表达的HLrg蛋白对细胞周期有调节作用,对肝癌细胞HepG2具有生长阻滞的作用。
Objective To stably overexpress HLrg in HepG2 hepatocellular carcinoma cell line and to observe the biological effects of overexpressed HLrg protein on HepG2 cell cycle and cell morphology. Methods The eukaryotic recombinant plasmid pcDNA3.1 (+) - hlrg was constructed and transfected into HepG2 cells for stable selection. The control group, pcDNA3.1 (+) - hlrg The experimental group was observed. Western blot analysis of HLrg protein. The cell cycle was measured by flow cytometry, the growth curve was measured by MTT method, and the change of cell morphology was observed by transmission electron microscopy. Results The HepG2 cell line stably overexpressing pcDNA3.1 (+) - hlrg was obtained. Compared with the control group, cell lines overexpressing HLrg showed G1 arrest, reduced microvilli, reduced mitotic phase, and slowed down the growth. Conclusion Stably overexpressed HLrg protein may play a role in the regulation of cell cycle and has a role in the growth arrest of HepG2 cells.