膜联蛋白A1对小鼠胰岛B细胞的调控作用

ANXA1 regulated proliferation of islet beta-cells in mice

目的探讨膜联蛋白A1(annexin A1,ANXA1)与胰岛B细胞株MIN6细胞增殖的关系。方法应用不同葡萄糖浓度(5.5mmol/L,11.1 mmol/L,16.7 mmol/L,25 mmol/L,33.3 mmol/L)刺激小鼠胰岛B细胞株MIN6细胞,通过CCK-8检测细胞增殖活性的变化,Western blot方法检测ANXA1蛋白的表达变化。经si RNA抑制MIN6细胞ANXA1表达后,分别采用流式细胞仪及Western blot方法观察细胞周期及细胞周期相关蛋白的变化。结果葡萄糖诱导的MIN6细胞增殖活性具有浓度依赖性,在葡萄糖浓度≤25 mmol/L时细胞增殖活性随葡萄糖浓度的增加而增加,葡萄糖浓度33.3 mmol/L时细胞增殖活性较25 mmol/L时下降。Anxa1蛋白表达与细胞增殖活性趋势相同。MIN6细胞转染Anxa1 si RNA后增殖活性下降,S期及G2/M期细胞明显减少,同时细胞周期相关蛋白(Cyclin D1、Cyclin E和CDK2)表达也明显下降。结论 Anxa1可正向调控细胞周期相关蛋白Cyclin D1、Cyclin E和CDK2的表达,进而影响MIN6细胞增殖活性。

Objective To study the relationship between Annexin 1（ANXA1） and proliferation of islet β-cells. Methods Stimulated by glucose with different concentrations（5.5 mmol/L, 11.1 mmol/L, 16.7 mmol/L, 25 mmol/L, 33.3 mmol/L）, the degree of cell proliferation of the mouse pancreatic islet beta cell line（MIN6） was assessed by CKK8 and the expression of ANXA1 protein was detected by Western blot. After inhibition of ANXA1 expression by small interfering RNA（ANXA1-si RNA） in MIN6 beta cell line, flow cytometry（FCM） method was used to analyze cell cycles and the expressions of cell cycle-related proteins were detected by Western blot. Results Glucose induced MIN6 cells proliferation in a concentration-dependent manner peaking at 25 mmol/L, while higher concentration（33.3 mmol/L） produced a minor decrease of proliferation comparing with 25 mmol/L. ANXA1 protein expression demonstrated a similar response to glucose. The cells proliferation decreased after transfection of ANXA1 si RNA by MIN6, and the percentage of cells in the S and G2/M phases also decreased significantly. Moreover, ANXA1 protein expression in MIN6 cells was positively related with Cyclin D1, Cyclin E and CDK2 proteins. Conclusion ANXA1 protein may affect proliferation of MIN6 cells by regulating Cyclin D1, Cyclin E and CDK2 protein.