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目的探讨氯化两面针碱对垂体腺瘤GH3细胞的抑制作用及其机制。方法不同浓度的氯化两面针碱作用于GH3细胞,培养不同时间后分别检测其作用效果:CCK-8检测各组GH3细胞存活率;Annexin V/PI凋亡试剂盒染色后,采用流式细胞术检测各组GH3细胞凋亡率;PI染色后采用流式细胞术进行细胞周期分析;实时定量PCR检测凋亡相关基因Bax、Bcl-2,以及细胞周期相关基因Cyclin B1、CDK1、p21和p27的mRNA表达水平;Western blotting法检测Bax、Bcl-2、Cyclin B1和CDK1蛋白,以及丝氨酸/苏氨酸激酶(AKT)和细胞外信号调节激酶(ERK)信号通路蛋白的表达水平。结果氯化两面针碱可抑制GH3细胞增殖,促进GH3细胞凋亡及细胞周期阻滞于G2/M期;氯化两面针碱能够抑制AKT及ERK信号通路的激活。结论氯化两面针碱有效抑制垂体腺瘤GH3细胞的增殖和促进其凋亡,可作为有效治疗垂体腺瘤的潜在药物。
Objective To investigate the inhibitory effect of nitidine chloride on pituitary adenoma GH3 cells and its mechanism. Methods Different concentrations of nitidine chloride acted on GH3 cells, and the effect of GH2 was detected after different time. The survival rate of GH3 cells in each group was detected by CCK-8. After the cells were stained with Annexin V / PI apoptosis kit, flow cytometry The apoptotic rates of GH3 cells in each group were detected by flow cytometry. Flow cytometry was used to analyze cell cycle after PI staining. Apoptosis-related genes Bax and Bcl-2, as well as cyclin B1, CDK1, p21 and p27 The mRNA expression of Bax, Bcl-2, Cyclin B1 and CDK1, and the expressions of serine / threonine kinase (AKT) and extracellular signal-regulated kinase (ERK) signaling pathways were detected by Western blotting. Results Nitidine chloride could inhibit the proliferation of GH3 cells and promote the apoptosis and cell cycle arrest of GH3 cells in G2 / M phase. Nitidine chloride could inhibit the activation of AKT and ERK signaling pathway. Conclusion Nitidine chloride can effectively inhibit the proliferation and promote the apoptosis of pituitary adenoma GH3 cells, which may be used as a potential drug for the treatment of pituitary adenoma.