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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。
Objective To investigate the effects of HBx-d382 and HBx-d431 deletion mutants on the biological behavior of immortalized QSG7701 hepatocytes. Methods The morphological changes of transfected cells were observed by HE staining. The biological characteristics of stable transfected cells were studied by MTT assay, soft agar colony formation assay, flow cytometry and tumorigenesis in nude mice. Results Compared with the pcDNA3 vector, the size and shape of HBx-2215 gene in HBx-d382 and HBx-d431 and HepG2.2.15 cell lines were different, Large, faster growth, high clonal formation rate (P <0.05). The percentages of S phase in pcDNA3 / HBx-d382 and pcDNA3 / HBx-2215 group were 32.8% and 35.0% higher than that in pcDNA3S (29.4%) and 13.1% / HBx-d431 group apoptosis rate (4.5%). Conclusion HBx-d382 and HBx-d431 deletion mutants can promote the proliferation and malignant transformation of QSG7701 cells.