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目的:探讨重构型caspase-3基因的表达对肿瘤细胞的凋亡诱导作用。方法:用重组PCR的方法获得大、小亚基顺序颠倒的重构型caspase-3基因并将其克隆入真核表达载体pcDNA3,转染乳腺癌细胞系SKBr3细胞,通过HE染色、流式细胞仪分析及免疫组化等方法观察其表达后对细胞的促凋亡活性,并将载体DNA直接注射荷瘤裸鼠研究其对肿瘤的抑制作用。结果:重构型caspase-3基因可在SKBr3细胞内表达,转染组较同期对照组出现明显的细胞死亡现象,pcDNA3-revcasp-3重组质粒直接注入对荷瘤裸鼠肿瘤生长有明显抑制作用。结论:重构型caspase-3基因表达可诱导SKBr3细胞凋亡。
Objective: To investigate the apoptosis-inducing effect of reconstructive caspase-3 gene on tumor cells. Methods: The reconstructed caspase-3 gene with inverted order of large and small subunits was obtained by recombinant PCR and cloned into the eukaryotic expression vector pcDNA3. The recombinant plasmid was transfected into breast cancer cell line SKBr3. HE staining and flow cytometry Instrument analysis and immunohistochemistry and other methods to observe the expression of its pro-apoptotic cells, and the carrier DNA directly into tumor-bearing nude mice to study its inhibition of the tumor. Results: The reconstructed caspase-3 gene was expressed in SKBr3 cells, and the cell death was observed in the transfected group compared with the control group. The direct injection of pcDNA3-revcasp-3 recombinant plasmid significantly inhibited tumor growth in nude mice . Conclusion: The reconstructed caspase-3 gene can induce the apoptosis of SKBr3 cells.