胰岛素瘤相关蛋白-2高表达对胰岛细胞增殖的影响

Effect of Autoantigen IA-2 Overexpression on the Proliferation of Insulinoma Cell Line RIN5F

目的:探讨1型糖尿病自身抗原胰岛素瘤相关蛋白-2(IA鄄2)高表达对胰岛细胞系RIN5F细胞增殖、生长周期和细胞凋亡的影响。方法:利用分子克隆技术构建IA鄄2真核表达载体,转染RIN5F细胞后筛选稳定高表达IA鄄2的细胞系,采用3鄄(4,5鄄二甲基)鄄5鄄(3鄄羧甲基苯环)鄄2鄄(4鄄硫基苯)鄄2H鄄四唑盐复合物(MTS)检测法确定细胞增殖速率,运用流式细胞术检测细胞生长周期和细胞凋亡水平。结果:IA鄄2高表达细胞增殖速率显著低于载体对照组细胞(P<0.05)。IA鄄2高表达细胞的细胞凋亡数显著高于载体对照组细胞(P<0.01)。IA鄄2高表达细胞中位于S期的细胞比率显著增加,而G2/M期的细胞比率减少(P<0.05)。结论:IA鄄2在胰岛细胞内高表达可抑制胰岛细胞的增殖,促进细胞凋亡,在G2鄄M期细胞生长阻滞。

Objective:To explore the role of autoantigen IA-2 overexpression to the proliferation, cell cycle, and apoptosis of insulinoma cell line RIN5F. Methods:The recombined IA-2 mammaline expression system constructed by gene cloning technique transfected the RIN5F cell by lipofectin and selected the stable overexpression IA-2 RIN5F cell line. Cell proliferations were measured by MTS one-solution kits. The cell cycle and cell apoptosis were analysed by flow cytometry after stained with PI. Results:The proliferation of IA-2 overexpression RIN5F cell line was lower than the control group(P < 0.05). The ratio of apoptosis of IA-2 overexpression RIN5F cell was significantly higher than the control group(P < 0.01). In the cell cycle of transcient transfect IA-2 RIN5F cell line, the ratio of S-period cell significantly increased than the control, but the G2/M period cell number decreased than the control(P < 0.05). Conclusion: The overexpression IA-2 can inhibit the cell proliferation and enhance the apoptosis. The overexpression IA-2 can also retard the cell in G2-M period.