【摘要】 目的:探讨蛋白激酶B(AKT)抑制剂卡帕塞替尼(AZD5363)对类风湿关节炎(RA)成纤维样滑膜细胞MH7A增殖、迁移和侵袭、凋亡以及自噬的影响。方法:将MH7A细胞随机分为对照组(无药物干预)、低浓度组(含3μmol/L AZD5363高糖培养基培养)、中浓度组(含6μmol/L AZD5363高糖培养基培养)和高浓度组(含9μmol/L AZD5363高糖培养基培养)。采用CCK-8法检测细胞增殖情况；采用流式细胞术和Transwell实验检测细胞凋亡、迁移和侵袭情况；采用Western blot检测细胞凋亡相关蛋白[B淋巴细胞瘤2(Bcl2)和Bcl2相关X蛋白(Bax)]、细胞迁移与侵袭相关蛋白[基质金属蛋白酶-1(MMP-1)和MMP-2]、自噬相关蛋白[自噬相关基因5(ATG5)、自噬相关蛋白7(ATG7)、微管相关蛋白1A/1B-轻链3(LC3Ⅱ/Ⅰ)、螯合体1(p62)]以及AKT、磷酸化AKT(p-AKT)蛋白表达。结果:MH7A细胞经过不同浓度AZD5363处理后，与对照组比较，随着药物浓度增加，细胞活力降低，细胞凋亡率增加，细胞迁移和侵袭率降低，Bcl2、MMP-1、MMP-2、ATG5、ATG7及LC3Ⅱ/Ⅰ蛋白表达降低，Bax、p62蛋白表达升高(均P<0.05)。结论:AZD5363可以明显抑制MH7A细胞增殖、迁移、侵袭及自噬，同时显著促进其凋亡。更多还原

【Abstract】 Objective:To investigate the effects of the protein kinase B(AKT) inhibitor capivasertib(AZD5363) on the proliferation, migration and invasion, apoptosis and autophagy in rheumatoid arthritis(RA) fibroblast-like synoviocyte MH7A.Methods:MH7A cells were randomly divided into control group(no drug intervention),low concentration group(cultured with 3 μmol/L AZD5363 in high glucose medium),medium concentration group(cultured with 6 μmol/L AZD5363 in high glucose medium) and high concentration group(cultured with 9 μmol/L AZD5363 in high glucose medium).Cell proliferation was detected by cell counting kit-8 assay.Apoptosis was detected using flow cytometry.Transwell assay was used to detect cell migration and invasion.Western blot assay was used to detect apoptosis-associated proteins B-lymphoblastoma-2 gene(Bcl2),Bcl2-Associated X protein(Bax),and cell migration and invasion-associated protein matrix metalloproteinase-1(MMP-1) and MMP-2,and autophagy-associated protein autophagy-associated gene 5(ATG5),ATG7,microtubule-associated protein 1A/1B-light chain 3(LC3Ⅱ/Ⅰ),chelator 1(p62),and AKT,phosphorylated AKT(p-AKT).Results:After MH7A cells were treated with different concentrations of AZD5363,compared with the control group, with the increase of drug concentration, the cell viability was decreased, the apoptosis rate was significantly increased, and the cell migration and invasion rate was decreased, and the protein expressions of Bcl2,MMP-1,MMP-2,ATG5,ATG7 and LC3Ⅱ/Ⅰ were decreased, and the protein expressions of Bax and p62 were increased(all P<0.05).Conclusion:AZD5363 can significantly inhibit the proliferation, migration, invasion and autophagy of MH7A cells, and significantly promote their apoptosis.更多还原