摘要
目的:探讨CD95 L在肝癌细胞凋亡过程中的作用. 方法:ELISA法对慢性乙型肝炎、肝炎肝硬化与肝癌患者血清可溶性CD95 L(sCD95L)水平进行了初步检测,构建了人CD95 L的重组真核表达体pcDNA3.1hisB-CD95 L, 将pcDNA3.1hisB-CD95 L转染至人肝癌细胞株HepG2细胞,采用Annexin V/PI双染后双变量流式细胞仪检测细胞凋亡率. 结果:sCD95L在肝癌患者明显低于肝炎及肝硬患者,构建的表达重组体pcDNA3.1hisB-CD95 L经菌落PCR和限制性酶切消化有预期的目的片段出现,DNA序列分析证实CD95 L完整、正确插入,转染后的HepG2细胞细胞凋亡率为36.30%;未转染CD95L的对照组细胞凋亡率11.53%. 结论:CD95L可使肝癌细胞凋亡.
AIM: To investigate CD95 ligand and its physiological function in liver neoplasms. METHODS: The levels of soluble Fas ligand (sFasL) were evaluated in a group of patients affected by hepatitis B virus (HBV)-induced chronic hepatitis, HBV-positive liver cirrhosis and hepatocellular carcinoma (HCC). To further study, we constructed recombinant eukaryotic expression vector pcDNA3.1 hisB-CD95L, which was then tranfected into human hepatoma cell line HepG2 by lipofection. After stained by annexin V and propidium iodine, HepG2 cells were detected by flow cytometer. RESULTS: s CD95L levels were significantly decreased in patients with HCC when compared to the patients with hepatitis or liver cirrhosis. The correct recombinant pcDNA3.1hisB-CD95L was selected by PCR and restriction endonuclease digestion and confirmed by DNA sequencing respectively. Subsequently a significant proportion of cells became apoptotic, as evidenced by positive annexin staining. CONCLUSION: CD95-CD95 ligand system can induce apoptosis of hepatoma cells.
出处
《世界华人消化杂志》
CAS
2004年第8期1789-1792,共4页
World Chinese Journal of Digestology
基金
湖南省卫生厅科研基金资助课题
No.9638~~
