HBx基因缺失型突变体对肝细胞QSG7701细胞增殖和凋亡的影响

目的研究HBx-d382和HBx-d431缺失型突变体对永生化QSG7701肝细胞生物学行为的影响。方法采用HE染色法,观察转染细胞形态学变化,通过MTT、软琼脂克隆形成实验、流式细胞仪及裸鼠成瘤实验研究稳定转染细胞的生物学特性。结果与转染空质粒pcDNA3相比,转染肝癌组织中HBx-d382和HBx-d431突变体及HepG2.2.15细胞株中HBx-2215基因的QSG7701细胞大小形态不一致、体积增大、核浆比例增大,生长速度更快,克隆形成率高（P〈0.05）。与转染空质粒pcDNA3S期百分比（29.4%）和凋亡率（13.1%）比较,pcDNA3/HBx-d382和pcDNA3/HBx-2215组细胞S期百分比比例增高,分别为32.8%和35.0%,pcDNA3/HBx-d431组凋亡率下降（4.5%）。结论 HBx-d382和HBx-d431缺失型突变体能促进QSG7701细胞增殖和恶性转化。

Effects of HBx gene deletion on the cell proliferation and apoptosis of the stem cell QSG7701

Objective To study the tumorigenicity of pcDNA3/HBx-d382 and pcDNA3/HBx-d431 and explore the biological effects of truncated HBx on QSG7701 cell line.Methods HE staining was used to observe the morphology of cells in each group.The biological characteristics of QSG7701 cell lines in each group were analyzed by MTT assay,soft agar colony formation assay,flow cytometry（FCM） and xenograft in nude mice.Results QSG7701 cell line transfected with pcDNA3/HBx-d382,pcDNA3/HBx-d431 or pcDNA3/HBx-2215 grew faster than the parent cell line.The colony formation rate was significantly higher in pcDNA3/HBx-d382,pcDNA3/HBx-d431 or pcDNA3/HBx-2215 transfected cells than in pcDNA3 transfected cells （18.27±1.35）%,（16.37±1.37）% and （15.40±1.44）% vs.（1.72±0.25）%,P0.05].Compared with pcDNA3 transfected cells,pcDNA3/HBx-d382 and pcDNA3/HBx-2215 transfected cells promoted more cells from G1 phase into S phase in cell cycle（29.4% vs.32.8% and 35.0% in S phase）.The apoptosis rate of pcDNA3/HBx-d431 QSG7701 cell line was singnificantly lower than control（13.1% vs.4.5%）.Conclusion Compare with HBx-2215（HBx of HepG2.2.15）,HBx-d382 and HBx-d431 could promote the proliferation of QSG7701 cells and have tumorigenic ability.