转染肝刺激因子基因增强肝癌细胞抗损伤作用研究

目的 将人肝刺激因子基因(hHSS)转染BEL-7402肝癌细胞,探讨其增强细胞抗损伤的作用。 方法 于肝癌细胞中转染hHSS,并以MTT法检测转染细胞活性;采用CCl4和H2O2损伤细胞,测定细胞死亡率和凋亡率,以及丝裂素活化蛋白激酶(MAPK)活性变化,观察转染细胞的抗损伤能力。 结果 hHSS表达的肝癌细胞增殖速度加快,同时转染hHSS细胞具有保护细胞免受两种毒剂的氧化损伤的能力,可以明显降低细胞死亡率及凋亡率[在CCl1-处理组和H2O2-处理组空载体对照凋亡细胞率分别为(32.44±0.52)％和(47.78±0.45)％,转染细胞凋亡率分别为(25.60±0.66)％和(37.40±0.69)％,t值分别为16.82和25.2,P<0.01],并活化酪氨酸信号分子MAPK。 结论 hHSS基因的功能与刺激细胞增殖有关。

Transfection of human hepatic stimulator substance gene could protect BEL-7402 cells against hepatotoxins

OBJECTIVE: To investigate protective effects of hHSS transfection against CCl4 or H2O2. METHODS: cDNA coding for hHSS was constructed into eukaryotic vector of pcDNA3.1 and transfected into BEL-7402 hepatoma cells. The expression of hHSS was analyzed with Northern blot. RESULTS: The growth of the hepatoma cells was remarkably enhanced 24 to 144h after hHSS gene transfection, which suggesting hHSS gene expression could stimulate cells activity. Meantime, incubation of both wild-type and vector-transfected as well as hHSS-transfected cells with CCl4 or H2O2 resulted in severe damage as marked by cell mortality and the rate of apoptosis. However, it appeared that the transfection of hHSS enabled the hepatoma cells to raise obvious resistance against CCl4 and H2O2 injury. Compared the vector cells to the vector-transfected cells, apoptosis ratio were (32.44+/-0.52)% and (25.60+/-0.66)% in which treated with CCl4, while (47.78+/-0.45)% and (37.40+/-0.69)% in which treated with H2O2, t value is 16.82 and 25.20, P<0.01. MAPK phosphorylation was also activated after HSS transfected. CONCLUSION: The function of hHSS gene expression could be related to proliferation of cell and protection against free radical damage.