circ_AFF2通过调节PI3K/AKT通路对类风湿关节炎滑膜成纤维细胞增殖、迁移和侵袭的影响

杨雯娣; 王峰

doi:10.13898/j.cnki.issn.1000-2200.2023.07.004

circ_AFF2通过调节PI3K/AKT通路对类风湿关节炎滑膜成纤维细胞增殖、迁移和侵袭的影响

杨雯娣,
王峰

Effect of circ_AFF2 on the proliferation, migration and invasion of rheumatoid arthritis synovial fibroblasts by regulating PI3K/AKT pathway

摘要

摘要:
目的探讨circ_AFF2通过调节磷脂酰肌醇3激酶(PI3K)/丝氨酸/蛋白激酶B(AKT)通路对类风湿关节炎(rheumatoid arthritis, RA)滑膜成纤维细胞的增殖、迁移、侵袭的作用机制。
方法采用qRT-PCR检测健康对照滑膜组织和RA滑膜组织中circ_AFF2相对表达量。通过体外培养RA滑膜成纤维MH7A细胞，将细胞分为si-NC组(转染si-NC)、si-circ_AFF2组(转染si-circ_AFF2)和si-circ_AFF2+IGF-1组(转染si-circ_AFF2后，采用50 ng/mL的PI3K/AKT通路激活剂IGF-1处理)。然后通过qRT-PCR检测各组细胞circ_AFF2相对表达量；MTT检测各组细胞活力；Western blotting检测各组细胞Ki-67、PCNA、MMP-2、E-cadherin、N-cadherin和PI3K/AKT通路相关蛋白的表达；Transwell检测各组细胞迁移和侵袭能力。
结果与健康对照滑膜组织相比，circ_AFF2相对表达量在RA滑膜组织中明显增加(P < 0.05)。与si-NC组相比，si-circ_AFF2组的circ_AFF2相对表达量、细胞活力、Ki-67以及PCNA蛋白表达均明显降低(P < 0.01)。与si-NC组相比，si-circ_AFF2组的迁移细胞数目、侵袭细胞数目、MMP-2以及N-cadherin蛋白表达均明显降低(P < 0.01)，E-cadherin蛋白表达明显增加(P < 0.01)。3组p-PI3K、p-AKT蛋白表达均明显降低(P < 0.01)，si-circ_AFF2组和si-circ_AFF2+IGF-1组p-PI3K、p-AKT蛋白表达均低于si-NC组(P < 0.05)，而3组PI3K、AKT蛋白表达无变化(P>0.05)。与si-circ_AFF2组相比，si-circ_AFF2+IGF-1组的circ_AFF2相对表达量、细胞活力、迁移细胞数目、侵袭细胞数目、Ki-67、PCNA、MMP-2及N-cadherin蛋白表达明显增加(P < 0.01)，E-cadherin蛋白表达明显降低(P < 0.01)。
结论circ_AFF2可以通过激活PI3K/AKT通路促进RA滑膜成纤维细胞的增殖、迁移和侵袭。

Abstract:
ObjectiveTo explore the mechanism of circ_AFF2 regulating the proliferation, migration, and invasion of rheumatoid arthritis (rheumatoid arthritis, RA) synovial fibroblasts by regulating the phosphatidylinositol 3-kinase (PI3K)/serine/protein kinase B (AKT) pathway.
MethodsqRT-PCR was used to detect the relative expression of circ_AFF2 in healthy control synovial tissue and RA synovial tissue.By culturing RA synovial fibroblast MH7A cells in vitro, the cells were divided into si-NC group (transfected with si-NC), si-circ_AFF2 group (transfected with si-circ_AFF2), and si-circ_AFF2+IGF-1 Group (after transfection of si-circ_AFF2, treated with 50 ng/mL PI3K/AKT pathway activator IGF-1).Then qRT-PCR was used to detect the relative expression of circ_AFF2 in each group of cells; MTT assay was used to detect cell viability in each group; Western blotting was used to detect the expression of Ki-67, PCNA, MMP-2, E-cadherin, N-cadherin, and PI3K/AKT pathway-related proteins in each group of cells; Transwell assay detects cell migration and invasion ability of each group.
ResultsCompared with healthy control synovial group, the relative expression of circ_AFF2 was significantly increased in RA synovial tissue (P < 0.01).Compared with the si-NC group, the relative expression of circ_AFF2, cell viability, Ki-67 and PCNA protein expressions in the si-circ_AFF2 group were significantly decreased (P < 0.01).Compared with the si-NC group, the number of migrating cells, the number of invasive cells, the expression of MMP-2 and N-cadherin protein in the si-circ_AFF2 group were significantly decreased (P < 0.01), and the protein expression of E-cadherin was significantly increased (P < 0.01).The protein expressions of p-PI3K and p-AKT in the three groups were significantly decreased (P < 0.01), and the protein expressions of p-PI3K and p-AKT in the si-circ_AFF2 group and the si-circ_AFF2+IGF-1 group were lower than those in the si-NC group (P < 0.05), while there was no change in the expression of PI3K and AKT among three groups (P>0.05).Compared with si-circ_AFF2 group, the relative expression of circ_AFF2, cell viability, number of migrating cells, number of invasive cells, Ki-67, PCNA, MMP-2 and N-cadherin protein expressions in si-circ_AFF2+IGF-1 group were significantly increased (P < 0.01), and E-cadherin protein expression was significantly decreased (P < 0.01).
ConclusionsCirc_AFF2 can promote the proliferation, migration and invasion of RA synovial fibroblasts by activating the PI3K/AKT pathway.

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