SHP-1基因在曲妥珠单抗致心肌细胞凋亡中的调控作用

发布时间：2018-08-06 14:38

【摘要】：目的初步观察转染SHP-1基因前后大鼠心肌H9C2细胞对抗曲妥珠单抗的能力,SHP-1蛋白表达情况及转染SHP-1基因对H9C2细胞增殖能力的影响。方法构建真核表达载体pc DNA3.1(+)-SHP-1和pc DNA3.1(+)-SHP-1C/S,利用脂质体转染法转染大鼠心肌H9C2细胞,经G418筛选阳性克隆,阳性克隆细胞在曲妥株单抗中维持生长。用Western blot检测SHP-1基因在大鼠心肌H9C2细胞中的稳定表达情况,并通过流式细胞学观察细胞凋亡曲线。结果蛋白水平证实,转染细胞内有SHP-1基因的表达,并且转染pc DNA3.1(+)-SHP-1的心肌细胞SHP-1表达明显升高,经曲妥珠单抗干预后蛋白表达量下降。转染pc DNA3.1(+)-SHP-1后H9C6细胞在曲妥珠单抗中凋亡率明显升高(P0.05)。结论向大鼠心肌H9C2细胞内转染SHP-1基因并使其稳定表达,可以促进大鼠心肌H9C2细胞的在曲妥珠单抗中的凋亡,为以后进一步探讨SHP-1基因的功能打下了基础。
[Abstract]:Objective to investigate the expression of SHP-1 protein in rat myocardial H9C2 cells before and after transfection of SHP-1 gene and the effect of SHP-1 gene transfection on the proliferation of H9C2 cells. Methods the eukaryotic expression vectors PC DNA3.1 () -SHP-1 and PC DNA3.1 () -SHP-1C / S were constructed and transfected into rat myocardial H9C2 cells by liposome transfection. The positive clones were screened by G418. Western blot was used to detect the stable expression of SHP-1 gene in rat myocardial H9C2 cells and the apoptosis curve was observed by flow cytometry. Results the expression of SHP-1 gene was confirmed in the transfected cells, and the expression of SHP-1 in the cardiomyocytes transfected with PC DNA3.1 () -SHP-1 was significantly increased, and the protein expression decreased after the intervention of trotozumab. After transfection of PC DNA3.1 () -SHP-1, the apoptosis rate of H9C6 cells increased significantly (P0.05). Conclusion transfection and stable expression of SHP-1 gene into rat myocardial H9C2 cells can promote the apoptosis of rat myocardial H9C2 cells in trotozumab and lay a foundation for further study on the function of SHP-1 gene.
【作者单位】：广东省东莞市人民医院肿瘤内科;南方医科大学南方医院肿瘤内科;南方医科大学中西医结合医院肿瘤中心;
【基金】：国家自然科学基金资助项目(编号:81272546) 广东省中医药局建设中医药强省科研立项课题(编号:20131072)
【分类号】：R965

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