新型hGLP-1基因对β-TC-6胰岛瘤细胞凋亡的影响

Research on the effect of a novel hGLP-1 analog gene in AXN-induced β-TC-6 cells

目的探讨新型人胰高血糖素样肽-1(hGLP-1)类似物基因(2×Val2-hGLP-1)对四氧嘧啶(AXN)诱导凋亡的β-TC-6胰岛瘤细胞株的影响。方法脂质体介导重组质粒pIRES2-EGFP/2×Val2-hGLP-1转染β-TC-6胰岛瘤细胞,荧光显微镜观察绿色荧光蛋白(GFP),酶联免疫吸附法(ELISA)检测目的蛋白表达;以AXN诱导细胞凋亡后观察细胞形态,Hoechst染色及四甲基偶氮唑盐光吸收法(WST-1)检测目的基因表达对细胞凋亡的影响。结果荧光显微镜观察到GFP表达;ELISA显示重组质粒转染组细胞培养液吸光度值为(2.53±0.05),高于其他研究组(P<0.05);细胞形态观察及Hoechst染色显示,AXN可诱导β-TC-6细胞凋亡;WST-1显示转染重组质粒的细胞组存活率为66.23%,高于其他研究组(P<0.05);Hoechst染色显示,经重组质粒转染的细胞凋亡减少。结论 2×Val2-hGLP-1的表达对胰岛β细胞凋亡具有一定抑制作用,为糖尿病基因治疗提供了相关试验研究基础。

Objective To investigate the effect of a novel hGLP-1 analog gene（2×Val2-hGLP-1）on alloxan（AXN）-induced apoptosis in β-TC-6 cells.Methods β-TC-6 cells were transfected with recombinant expression plasmid pIRES2-EGFP/2×Val2-hGLP-1 by cationic liposome.The expression of green fluorescent protein（GFP）was observed by fluorescent microscopy,and of hGLP-1 in the cultural media was determined by enzyme linked immunosorbent assay（ELISA）.After being induced by,the effect of 2×Val2-hGLP-1 on β-TC-6 cells were determined by microscope observation,Hoechst staining and MTT assay.Results The expression of GFP were obtained.The optical density of cultural media from recombinant plasmid transfected group was 2.53±0.05,and higher than other groups（P0.05）.Morphologic observation and Hoechst staining confirmed the apoptosis of β-TC-6 cells,induced by AXN.MTT assay indicated that the survival rate of recombinant plasmid transfected group（66.23%）was higher than other groups（P0.05）.Conclusion The expressioin of 2×Val2-hGLP-1 could inhibit the apoptosis of β-TC-6 cells,which lays an experimental foundation for the gene therapy of diabetes mellitus.