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microRNA-448对类风湿关节炎滑膜细胞增殖和凋亡的影响及其机制研究
作者:徐立  张旭然  张淼 
单位:阜新市中心医院 骨外科, 辽宁 阜新 123000
关键词:类风湿关节炎滑膜细胞 microRNA-448 凋亡 增殖 程序性细胞死亡因子5 
分类号:R392.12
出版年·卷·期(页码):2021·49·第十一期(1290-1296)
摘要:

目的:探讨microRNA-448(miR-448)对类风湿关节炎滑膜细胞增殖和凋亡的影响及其机制。方法:类风湿关节炎滑膜细胞MH7A分成Control组、Anti-NC组(转染inhibitor control)、Anti-miR-448组(转染miR-448 inhibitor)、Anti-miR-448+si-NC组(共转染miR-448 inhibitor、siRNA control)、Anti-miR-448+si-程序性细胞死亡因子5(PDCD5)组(共转染miR-448 inhibitor、PDCD5 siRNA)。CCK-8检测细胞增殖,PI单染法检测细胞周期,Annexin V-FITC/PI双染法检测细胞凋亡,蛋白质印迹法检测C-Caspase-3、cyclin D1、p21蛋白表达。荧光素酶报告系统鉴定miR-448和PDCD5的靶向关系。结果:与Control组和Anti-NC组比较,Anti-miR-448组细胞存活率降低[(100.00±9.52)%、(99.71±10.36)% vs(52.62±4.47)%,P<0.05],凋亡率升高[(4.32±0.35)%、(4.18±0.62)% vs(12.51±1.32)%,P<0.05],G0/G1期比例升高[(48.51±3.26)%、(50.80±6.20)% vs(63.81±5.41)%,P<0.05],细胞中C-Caspase-3、p21蛋白表达水平升高,cyclin D1蛋白表达水平下降。与Anti-miR-448+si-NC组比较,Anti-miR-448+si-PDCD5组类风湿关节炎滑膜细胞存活率升高[(100.00±10.35)% vs(147.84±16.50)%,P<0.05],细胞G0/G1期比例降低[(64.08±6.10)% vs(53.11±4.75)%,P<0.05],细胞凋亡率降低[(13.05±1.16)% vs(6.92±0.35)%,P<0.05],细胞中C-Caspase-3、p21、PDCD5蛋白表达水平降低,cyclin D1蛋白表达水平升高。结论:下调的miR-448通过靶向上调PDCD5表达抑制类风湿关节炎滑膜细胞增殖,阻滞细胞周期,诱导细胞凋亡。

Objective: To investigate the effects of microRNA-448 (miR-448) on the proliferation and apoptosis of synovial cells in rheumatoid arthritis and its mechanism. Methods: Rheumatoid arthritis synovial cells MH7A were divided into Control group, Anti-NC group (transfected with inhibitor control), Anti-miR-448 group (transfected with miR-448 inhibitor), Anti-miR-448+si-NC group (co-transfected with miR-448 inhibitor, siRNA control), Anti-miR-448+si-PDCD5 group (co-transfected with miR-448 inhibitor, PDCD5 siRNA). Cell proliferation was detected by CCK-8, cell cycle was detected by PI single staining, cell apoptosis was detected by Annexin V-FITC/PI double staining, and protein expressions of C-Caspase-3, cyclin D1 and p21 were detected by Western blot. The luciferase reporting system was used to identify the targeting relationship between miR-448 and PDCD5. Results: Compared with Control group and Anti-NC group, Anti-miR-448 group cell survival rate decreased[(100.00±9.52)%,(99.71±10.36)% vs (52.62±4.47)%,P<0.05], apoptosis rate increased[(4.32±0.35)%,(4.18±0.62)% vs (12.51±1.32)%,P<0.05], G0/G1 phase ratio increased[(48.51±3.26)%,(50.80±6.20)% vs (63.81±5.41)%,P<0.05], and C-Caspase-3 and p21 protein increased, and the expression level of cyclin D1 protein decreased. Compared with the Anti-miR-448+si-NC group, the survival rate of rheumatoid arthritis synovial cells in the Anti-miR-448+si-PDCD5 group was increased [(100.00±10.35)% vs (147.84±16.50)%, P<0.05], cell G0/G1 phase ratio decreased [(64.08±6.10)% vs (53.11±4.75)%, P<0.05], cell apoptosis rate decreased [(13.05±1.16)% vs (6.92±0.35)%, P<0.05], the expression levels of C-Caspase-3, p21 and PDCD5 protein decreased, and the expression level of cyclin D1 protein increased. Down-regulation of miR-448 targeted up-regulation of PDCD5 expression. Conclusion: Down-regulation of miR-448 inhibits the proliferation of rheumatoid arthritis synovial cells, blocks the cell cycle and induces apoptosis by targeting the up-regulation of PDCD5 expression.

参考文献:

[1] 黄皆和,郑文标,管军辉,等.白芍总苷对类风湿性关节炎的治疗作用及Wnt/β-catenin通路的影响[J].中国药师,2020,23(2):218-223.
[2] 白冰清,辛芳冉,魏婷婷,等.TNF-α-308G/A基因多态性与TNF-α抑制药治疗类风湿关节炎疗效相关性的Meta分析[J].药物流行病学杂志,2019,28(4):215-222.
[3] HAM S,BAE J B,LEE S,et al.Epigenetic analysis in rheumatoid arthritis synoviocytes[J].Exp Mol Med,2019,51(2):1-13.
[4] HOU C,WANG D,ZHANG L.MicroRNA-34a-3p inhibits proliferation of rheumatoid arthritis fibroblast-like synoviocytes[J].Mol Med Rep,2019,20(3):2563-2570.
[5] 谭佳,肖晨,陈金娜,等.miRNA调控线粒体功能的研究进展[J].生命的化学,2020,40(2):250-255.
[6] 旷达彬,左美玲,郑艳平.miRNA海绵在疾病中的研究进展[J].中南药学,2018,16(12):1733-1739.
[7] WU X,TANG H,LIU G,et al.miR-448 suppressed gastric cancer proliferation and invasion by regulating ADAM10[J].Tumor Biol,2016,37(8):10545-10551.
[8] WANG Y,PANG Q J,LIU J T,et al.Down-regulated miR-448 relieves spinal cord ischemia/reperfusion injury by up-regulating SIRT1[J].Braz J Med Biol Res,2018,51(5):e7319.
[9] YANG Z C,QU Z H,YI M J,et al.MiR-448-5p inhibits TGF-β1-induced epithelial-mesenchymal transition and pulmonary fibrosis by targeting Six1 in asthma[J].J Cell Physiol,2019,234(6):8804-8814.
[10] JIN F,HU H,XU M,et al.Serum microRNA profiles serve as novel biomarkers for autoimmune diseases[J].Front Immunol,2018,16(9):2381-2389.
[11] 陈占昆,吕厚山,王宁.PDCD5在类风湿关节炎成纤维样滑膜细胞凋亡中表达上调[J].中国生物化学与分子生物学报,2008,24(6):563-568.
[12] BABOSOVA O,KAPRALOVA K,RASKOVA K L,et al.Iron chelation and 2-oxoglutarate-dependent dioxygenase inhibition suppress mantle cell lymphoma's cyclin D1[J].J Cell Mol Med,2019,23(11):7785-7795.
[13] OU W B,NI N,ZUO R,et al.Cyclin D1 is a mediator of gastrointestinal stromal tumor KIT-independence[J].Oncogene,2019,38(39):6615-6629.
[14] PALAIOLOGOS P,CHRYSIKOS D,THEOCHARIS S,et al.The prognostic value of G1 cyclins,p21 and Rb protein in patients with colon cancer[J].Anticancer Res,2019,39(11):6291-6297.
[15] BERNARD A,CHEVRIER S,BELTJENS F,et al.Cleaved Caspase-3 transcriptionally regulates angiogenesis-promoting chemotherapy resistance[J].Cancer Res,2019,79(23):5958-5970.
[16] ZENG C Y,LI C G,SHU J X,et al.ATP induces caspase-3/gasdermin E-mediated pyroptosis in NLRP3 pathway-blocked murine macrophages[J].Apoptosis,2019,24(9-10):703-717.
[17] KUO W T,SHEN L,ZUO L,et al.Inflammation-induced occludin downregulation limits epithelial apoptosis by suppressing Caspase-3 expression[J].Gastroenterology,2019,157(5):1323-1337.
[18] LABI V,PENG S,KLIRONOMOS F,et al.Context-specific regulation of cell survival by a miRNA-controlled BIM rheostat[J].Genes Dev,2019,33(23-24):1673-1687.
[19] AHMAD M,JALIL F,HAQ M,et al.Effect of variation in miRNA-binding site (rs8176318) of the BRCA1 gene in breast cancer patients[J].Turk J Med Sci,2019,49(5):1433-1438.
[20] WANG K,ZHANG X,WANG Y,et al.PDCD5 regulates iNKT cell terminal maturation and iNKT1 fate decision[J].Cell Mol Immunol,2019,16(9):746-756.
[21] 陈占昆,王宁,吕厚山.程序化细胞死亡因子5过表达促进雷公藤内醇酯诱导的类风湿关节炎成纤维样滑膜细胞凋亡[J].北京大学学报(医学版),2008,40(6):567-571.

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