局解手术学杂志2023，Vol.32Issue(12)：1029-1034,6.DOI:10.11659/jjssx.10E022066
Epac1对垂体腺瘤细胞增殖和细胞周期的影响及分子机制研究
Effects and molecular mechanisms of Epac1 on pituitary adenoma cell proliferation and cell cycle
摘要
Abstract
Objective To explore the molecular mechanism of pituitary growth hormone adenoma cell proliferation.Methods Functional growth hormone-secreting pituitary adenoma(fGH-PA)tissue samples were collected from 12 patients with acromegaly.The exchange protein 1 directly activated by cAMP(Epac1)mRNA expression levels in fGH-PA tissues and rat RGC-5,MMQ and GH3 cells were determined by qPCR.The expression levels of Epac1 in fGH-PA tissues were determined by immunohistochemistry.Western blot was used to determine the expression levels of Epac1 in MMQ and GH3 cells.Overexpression or knockdown of Epac1,or knockdown of cAMP response element-binding protein(CREB)in GH3 cells,cell cycle changes were determined by flow cytometry,cell proliferation ability was determined by CCK-8 assay,and the expression levels of p-CREB,CREB,Cyclin D1,CDK2 and p21 in cells were determined by Western blot.Results qPCR and immunohistochemistry results showed that the expression levels of Epac1 mRNA and protein in fGH-PA tissues were significantly higher than those in adjacent normal tissues(P<0.05).qPCR and Western blot results showed that compared with RGC-5 cells,the expression levels of Epac1 mRNA and protein in MMQ and GH3 cells were significantly increased(P<0.05).After overexpres-sion of Epac1 in GH3 cells,compared with the Control group,the proportion of cells in G0/G1 phase and S phase in the Epac1-OE group were significantly reduced(P<0.05),and the proportion of cells in G2/M phase were significantly increased(P<0.05);the cell prolifera-tion ability were significantly enhanced(P<0.05);the expression levels of p-CREB and Cyclin D1 in cells were significantly increased(P<0.05),the expression levels of CDK2 and p21 were significantly decreased(P<0.05),while there was no significant change in the expression level of CREB between the two groups(P>0.05).After knockdown of Epac1 or knockdown of CREB in GH3 cells,all of the above results were reversed(P<0.05).Conclusion The overexpression of Epac1 in pituitary growth hormone adenoma cells can up-regulate the levels of p-CREB in cells,and promote adenoma cells to pass through G1/S phase checkpoint,resulting in cell cycle checkpoint disorder and massive proliferation,but it does not affect the expression levels of CREB.关键词
功能性生长激素垂体腺瘤/细胞增殖/细胞周期/G1/S期检查点蛋白/Epac1/CREBKey words
functional growth hormone-secreting pituitary adenoma/cell proliferation/cell cycle/G1/S phase checkpoint protein/Epac1/CREB分类
医药卫生引用本文复制引用
孙炜,王俊文,李然,黄逸民,张卓,雷霆..Epac1对垂体腺瘤细胞增殖和细胞周期的影响及分子机制研究[J].局解手术学杂志,2023,32(12):1029-1034,6.基金项目
国家自然科学基金(82173136) （82173136）
