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Hk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 stable over-express HLrg in HepG2 cells and observe its influence on cell cycle and cell morphology. Methods Eukaryotic expression vector pcDNA3.1 ( + )- hlrg was constructed, and then transfected into HepG2 cells. The HLrg protein was detected by Western blotting. Cell cycle was surveyed by FCM; cell morphology was observed by TEM; cell growth curve was detected by M'IT analysis. Results Cell strains of stable over-expression Hlrg HepG2-pcDNA3.1( + )-hlrg was obtained. As comparing to the control, cells of stable over-expression Hlrg was blocked in G1 phase. Meanwhile, microvillus of the cells decreased, fission percentages of the cells reduced, and growth of the cells was put off. Conclusion Over-expression of HLrg protein could regulate cell cycle of HepG2 cells and block growth of HepG2 cells.