CircPTTG1IP调控miR-223-3p/SMAD3轴对类风湿关节炎成纤维样滑膜细胞增殖、迁移和炎症反应的影响-现代医学

CircPTTG1IP调控miR-223-3p/SMAD3轴对类风湿关节炎成纤维样滑膜细胞增殖、迁移和炎症反应的影响

单位：1. 湖北省十堰市太和医院(湖北医药学院附属医院) 中西医结合科, 湖北十堰 442000;
2. 十堰市人民医院中医科, 湖北十堰 442000

关键词：Circ-PTTG1IP miR-223-3p 类风湿关节炎成纤维样滑膜细胞增殖迁移炎症反应

分类号：R593.22

出版年·卷·期（页码）：2023·51·第四期（454-461）

摘要：

目的:探讨CircPTTG1IP调控miR-223-3p/SMAD3轴对类风湿关节炎(RA)成纤维样滑膜(FLS)细胞增殖、迁移和炎症反应的影响。方法:实时荧光定量聚合酶链反应(qRT-PCR)、蛋白质印迹法(Western blot)检测正常FLS细胞和RA-FLS细胞中CircPTTG1IP、miR-223-3p表达以及SMAD3蛋白表达；双荧光素酶实验验证Circ-PTTG1IP与miR-223-3p、miR-223-3p与SMAD3的关系。将RA-FLS细胞分为RA-FLS组、si-NC组、si-PTTG1IP组、si-PTTG1IP+anti-NC组、si-PTTG1IP+anti-miR-223-3p组，qRT-PCR检测RA-FLS中Circ-PTTG1IP、miR-223-3p表达；CCK8检测RA-FLS增殖情况；流式细胞术检测RA-FLS凋亡率；Transwell 实验检测RA-FLS细胞迁移及侵袭情况；Western blot 检测SMAD3、EMT相关蛋白表达；ELISA法检测IL-2、IL-6水平。结果:与FLS组比较，RA-FLS组CircPTTG1IP、SMAD3上调，miR-223-3p下调；与RA-FLS组、si-NC组比较，si-PTTG1IP组OD值，RA-FLS细胞迁移、侵袭数量，Circ-PTTG1IP、SMAD3、N-cadherin、Vimentin蛋白表达，IL-6、MMP-2水平均显著降低(P<0.05)，miR-223-3p表达、细胞凋亡率、E-cadherin蛋白表达以及IL-2水平均显著升高(P<0.05)；miR-223-3p沉默减轻了上述指标的变化；Circ-PTTG1IP靶向调控miR-223-3p/SMAD3轴。结论:沉默Circ-PTTG1IP可能通过上调miR-223-3p来抑制SMAD3表达，进而抑制RA-FLS增殖、迁移和炎症反应。

Objective: To investigate the impacts of CircPTTG1IP on the proliferation, migration and inflammatory response of fibroblast like synovial(FLS) cells in rheumatoid arthritis(RA) by regulating miR-223-3p/SMAD3 axis. Methods: Real-time quantitative fluorescence polymerase chain reaction(qRT-PCR) and western blot were used to detect the expression of CircPTTG1IP, miR-223-3p and SMAD3 protein in normal FLS cells and RA-FLS cells. The relationships between Circ-PTTG1IP and miR-223-3p, miR-223-3p and SMAD3 were verified by dual luciferase assay. RA-FLS cells was divided into RA-FLS group, si-NC group, si-PTTG1IP group, si-PTTG1IP+anti-NC group, and si-PTTG1IP+anti-miR-223-3p group. The expression of CircPTTG1IP and miR-223-3p in RA-FLS was detected by qRT-PCR; CCK8 was used to detect the proliferation of RA-FLS cells; flow cytometry was used to detect the apoptosis rate of RA-FLS cells; Transwell assay was used to detect migration and invasion of RA-FLS cells; Western blot was used to detect the expression of SMAD3 and EMT related proteins; ELISA was used to detect the levels of IL-2 and IL-6. Results: Compared with FLS group, CircPTTG1IP and SMAD3 were up-regulated, and miR-223-3p was down-regulated in RA-FLS group; Compared with RA-FLS group and si-NC group, the OD value, the numbers of RA-FLS cells migration and invasion, CircPTTG1IP, SMAD3, N-cadherin, Vimentin protein expression, IL-6 and MMP-2 levels in si-PTTG1IP group were significantly decreased(P<0.05), the expression of miR-223-3p, apoptosis rate, the protein expression of E-cadherin and the level of IL-2 were significantly increased(P<0.05); miR-223-3p silencing alleviated the changes of the above indicators; CircPTTG1IP targeted and regulated the miR-223-3p/SMAD3 axis. Conclusion: Silencing CircPTTG1IP may inhibit the expression of SMAD3 by up-regulating miR-223-3p, thereby inhibiting the proliferation, migration and inflammatory response of RA-FLS.

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