摘要
为了解α1,3-岩藻糖基转移酶Ⅶ(α1,3FucT-Ⅶ)转染对全反式维甲酸诱导的人肝癌7721细胞凋亡的影响及其分子机理,本研究通过质粒转染把α1,3FucT-Ⅶ导入人肝癌7721细胞,建立了稳定表达不同量α1,3FucT-Ⅶ的H7721细胞株。采用流式细胞术测定细胞表面SLex的表达及细胞凋亡、Westernblot方法检测Caspase-3、免疫荧光染色法观测细胞骨架蛋白---肌动蛋白F-actin的变化、caspase-3抑制剂DEVD-CHO抑制caspase-3的活化。研究结果表明,用全反式维甲酸诱导细胞凋亡,α1,3FucT-Ⅶ的过量表达使凋亡细胞数量明显增多,caspase-3的活性和细胞骨架F-actin的破坏均增高,抑制剂DEVD-CHO可明显抑制细胞凋亡,但F-actin的破坏程度并未得到改善。通过caspase-3通路转染α1,3FucT-Ⅶ增加了肝癌细胞对全反式维甲酸诱导的凋亡的敏感性,从而为肝癌的防治提供了实验依据。
To study the effect of α1,3 fucosyltransferase Ⅶ （α1,3FucT-Ⅶ） transfection on the apoptosis of human hepatocarcinoma cells induced by all trans retinoic acid （ATRA） and its molecular mechanism. The cDNA of α1,3 FucT-Ⅶ was transfected into human H7721 hepatocarcinoma cell line, and the stable cell lines with different expressions of α1,3 FucT-Ⅶ were established. And cell surface SLex and cell apoptosis were detected by flow-cytometry. The expressions of caspase-3 were examined using western blot. The cell skeleton protein F-actin was observed by immunofluorescence staining. DEVD-CHO was used to inhibit the activation of caspase-3. Using the all trans retinoic acid （ATRA） to induce cell apoptosis, we found the apoptotic cell numbers and activated fragments of caspase-3 in cells with overexpressed α1,3 FucT- Ⅶ gene were increased dramatically. Again, the structure of F-actin in α1,3 FucT- Ⅶ transfected cells was disturbed compared with control cells. DEVD-CHO could inhibit cell apoptosis in α1,3 FucT-Ⅶ transfected cells, but the disturbance of F-actin structure was not restored.The transfection of α1,3 FucT-Ⅶ increases the susceptibility of ATRA induced cell apoptosis that mediated by caspase-3, provides the experimental basis for the prevention and treatment of liver cancer.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2009年第4期709-714,共6页
Genomics and Applied Biology
基金
广西自然科学基金资助项目(桂科能05112001-4C)
