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目的探讨活性氧自由基对骨髓源性肝前体细胞恶性转化的影响。方法取经过鉴定的3代骨髓间充质干细胞(BMSCs)分为3组:对照组(BMSCs),生长因子诱导组(BMSCs+HGF+EGF),过氧化氢刺激组(BMSCs+HGF+EGF+H2O2)。分别在细胞培养的第7天、14天、21天、28天用RT-PCR方法检测各组细胞的白蛋白(ALB)、甲胎蛋白(AFP)、角蛋白18(CK18)mRNA的表达。用免疫细胞化学方法检测细胞γ-谷氨酰转酞酶(γ-GT)的表达,用Schiff染色方法检测细胞DNA含量,用流式细胞仪检测非整倍体细胞。结果生长因子诱导组及过氧化氢刺激组细胞有ALB mRNA、AFP mRNA及CK18 mRNA的表达,过氧化氢刺激组细胞AFP mRNA、CK18mRNA、!-GT的表达、DNA含量及非整倍体细胞均高于生长因子诱导组(P<0.05)。结论骨髓间充质干细胞在体外诱导成为肝前体细胞,活性氧自由基可促使肝前体细胞过度增殖并恶性转化。
Objective To investigate the effect of reactive oxygen species (ROS) on the malignant transformation of bone marrow-derived liver progenitor cells. Methods BMSCs of the 3rd generation were identified and divided into 3 groups: control group (BMSCs), BMSCs + HGF + EGF group, BMSCs + HGF + EGF + H2O2). The expression of ALB, AFP and CK18 mRNA in each group were detected by RT-PCR on day 7, day 14, day 21 and day 28 respectively. The expression of γ-glutamyl transpeptidase (γ-GT) was detected by immunocytochemistry, the DNA content was detected by Schiff staining and the aneuploid cells were detected by flow cytometry. Results The expression of ALB mRNA, AFP mRNA and CK18 mRNA in cells induced by growth factor and hydrogen peroxide stimulated cells, the expression of AFP mRNA, CK18 mRNA and! - GT, the content of DNA and aneuploid cells Higher than the growth factor induction group (P <0.05). Conclusion Bone marrow-derived mesenchymal stem cells are induced to become precursor cells of liver in vitro. Active oxygen free radicals can promote the proliferation and malignant transformation of liver precursor cells.