摘要
目的:构建胰岛素样生长因子结合蛋白7(insulin-like growth factor binding protein7,IGFBP7)表达质粒(pEGFC1-IGFBP7),研究IGFBP7对恶性黑素瘤细胞SK-MEL-28凋亡的影响。方法:构建pEGFC1-IGFBP7质粒,并将pEGFC1-IGFBP7质粒及空质粒分别转染入SK-MEL-28细胞,荧光显微镜观测细胞转染效率,Annexin-FITC/PI检测转染后SK-MEL-28细胞的凋亡。结果:成功构建pEGFC1-IGFBP7质粒,用Effectene试剂能将pEGFC1-IGFBP7质粒有效转染入SK-MEL-28细胞,转染效率达61%。流式细胞仪结果显示pEGFC1-IGFBP7可明显促进SK-MEL-28细胞凋亡,转染24h后的凋亡率达(28.4±2.57)%,转染空质粒及未转染组细胞凋亡率分别为(5.8±0.44)%和(6.4±0.71)%(F=406.138,P<0.05)。结论:pEGFC1-IGFBP7能有效诱导黑素瘤SK-MEL-28细胞凋亡,为IGFBP7为基础的黑素瘤基因治疗提供了实验依据。
Objective：To construct the insulin-like growth factor binding protein 7 （IGFBP7） expression plasmid （pEGFC1-IGFBP7） and to investigate the effect of IGFBP7 on the apoptosis of SK-MEL-28 （human malignant melanoma cell line） cells.Methods：The pEGFC1-IGFBP7 plasmid was constructed;pEGFC1-IGFBP7 and empty plasmids were transfected into SK-MEL-28 cells separately.The transfection efficiency was observed under fluorescence microscope.Apoptosis of SK-MEL-28 cells after transfection was detected by Annexin-FITC/PI staining.Results：The pEGFC1-IGFBP7 plasmid was successfully constructed and was effectively transfected into SK-MEL-28 cells by Effectene reagent,with the transfection rate being 61%.The results of flow cytometry showed that pEGFC1-IGFBP7 significantly induced apoptosis of SK-MEL-28 cells,with the apoptotic rates of pEGFC1-IGFBP7,empty vector,and non-transfected plasmid groups being （28.4±2.57）%,（5.8±0.44）%,and （6.4±0.71）% 24 h after transfection,respectively （F=406.138,P〈0.05）.Conclusion：pEGFC1-IGFBP7 can effectively induce apoptosis of malignant melanoma SK-MEL-28 cells,which provides an experimental basis for IGFBP7 gene-based therapy of malignant melanoma.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2010年第1期36-39,共4页
Chinese Journal of Cancer Biotherapy
基金
国家教育部新教师基金资助项目(No.20070487140)~~
