重组抗HER2 ScFv／tBid基因的表达对SGC7901胃癌细胞的促凋亡作用

Investigation of Apoptosis of the SGC7901 Cells Induced by the Expression of the Recombinant Gene of anti-HER2 ScFv/tBid

目的:构建重组抗HER2 ScFv／tBid载体并观察其对胃癌SGC7901细胞的促凋亡作用。方法:将重组抗HER2 ScFv／tBid基因克隆入真核表达载体pCMV中,转染SGC7901细胞,用RT- PCR方法检测目的基因在mRNA水平的表达,间接免疫荧光法检测目的蛋白表达和细胞形态学变化,通过细胞计数检测转染目的基因后对细胞生长的影响,通过检测细胞周期来观察其促凋亡作用。结果:转染SGC7901细胞后,检测出目的蛋白的表达。细胞计数发现细胞的增殖被明显抑制。细胞周期分析有明显的凋亡峰出现,说明重组抗HER2 ScFv／tBid表达后有促凋亡作用。结论:重组抗HER2 ScFv／tBid基因可以在转染的SGC7901细胞中表达,并且可抑制转染细胞的生长,诱导细胞发生凋亡。

Objetive ： To investigate whether apoptosis of SGC7901 cells can be induced by the expression of the recombinant gene of anti-HER2 ScFv/tBid. Methods： The recombinant anti-HER2 ScFv/tBid gene was cloned into vector pCMV and the recombinant plasmid was transfected into SGC7901 cells. The gene expression was detected by RT-PCR and immunofluorescent staining. Cell counting was carried out to show the effect of the gene transfection on cell growth. At the same time, significant apoptotic peak was detected by flow cytometry in recombinant anti-HER2 ScFv/tBid gene transfected ceils. Results： The fusion protein of anti-HER2 ScFv/tBid was observed in the cytoplasm of transfected SGC7901 ceils. The transfected ceils displayed typical cell growth inhibition and apoptosis. Conclusion ： Fusion protein of anti-HER2 ScFv/tBid can induce apoptosis of SGC7901.