摘要
目的 :探讨具有自发活性的caspase 3分子对乳腺癌肿瘤细胞凋亡的诱导作用。方法 :重构型caspase 3转染SKBr 3细胞后 ,以观察细胞的形态、进行细胞计数和及流式细胞仪分析等方法 ,验证caspase 3大、小亚基顺序颠倒的重构型caspase 3分子的促凋亡活性。将重组真核表达载体 pcDNA3 revcas pase 3直接注射荷瘤裸鼠研究其对肿瘤生长的抑制作用 ,间接免疫荧光染色法检测肿瘤组织中重构型caspase 3基因表达情况 ,用TUNEL法检测肿瘤细胞的凋亡。结果 :重构型cas pase 3基因转染后 ,可诱导SKBr 3细胞凋亡 ,将 pcDNA3 revcaspase 3重组质粒直接注射荷SKBr 3瘤裸小鼠后 ,肿瘤的生长明显受到抑制 ,免疫组化染色可检出重构型caspase 3蛋白的表达 ;TUNEL法检测证实肿瘤细胞发生凋亡。结论 :重构型caspase 3基因的体内表达可诱导SKBr
AIM: To explore the apoptosis-inducing effect of a constitutively active caspase-3 molecule on human breast carcinoma SKBr-3 cells. METHODS: A revcaspase-3 gene was generated by arranging the coding sequence of the small subunit preceding that of the large one, i.e. in a reverse order of the wild-type caspase-3 gene. A pcDNA3-revcaspase-3 plasmid was constructed and transfected into SKBr-3 cells, and the proapoptotic effect of revcaspase-3 was examined by morphological observation of the cells, cell counting and flow cytometry analysis. The in vivo tumor suppression was further evaluated following i.t. administration of pcDNA3-revcaspase-3, while the apoptosis in tumor tissues was assayed using TUNEL. RESULTS: The apoptosis of SKBr-3 cells was induced after introduction of the revcaspase-3 gene, and tumor growth was strongly suppressed following injection of the pcDNA3 vector harboring revcaspase-3 gene. Revcaspase-3 was expressed in the cells of tumor tissues and apoptosis of the tumor cells was confirmed by TUNEL assays. CONCLUSION: Apoptosis of SKBr-3 cells in a mouse tumor model could be induced by in vivo expression of a revcaspase-3 gene.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第1期23-26,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家高技术研究发展计划 (863)资助 (No.2 0 0 1AA2 1 71 0 1 )
国家杰出青年科学基金资助 (No .3992 50 36)
军队杰出中青年人才研究基金资助 (No .98J0 0 9)
