miR-203a-5p调节Bcl-2/beclin-1通路对瘢痕疙瘩成纤维细胞增殖、迁移和凋亡的影响

Effect of miR-203a-5p on the proliferation,migration and apoptosis of keloid fibroblasts by regulating Bcl-2/beclin-1 pathway

目的探讨miR-203a-5p对瘢痕疙瘩成纤维细胞增殖、迁移和凋亡的影响。方法2020年1月至12月筛选靶向B细胞淋巴瘤/白血病-2(B-cell lymphoma/leukemia-2,Bcl-2)的微小核糖核酸(microRNA,miRNA),对瘢痕疙瘩成纤维细胞(keloid fibroblasts,KFs)进行转染,检测细胞增生、迁移和凋亡情况,同时检测细胞中Bcl-2、Bcl-2相关X蛋白(Bcl-2 related X protein,Bax)半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、beclin-1、自噬相关蛋白微管相关蛋白1轻链3Ⅱ(microtubule-associated-protein light-chain-3,LC3Ⅱ)、胶原蛋白Ⅰ(collagenⅠ)蛋白表达水平。结果miR-203a-5p与Bcl-2有潜在的结合位点;瘢痕疙瘩中miR-203a-5p mRNA低于正常皮肤、Bcl-2 mRNA高于正常皮肤(P<0.05)。miR-203a-5p抑制剂组KFs细胞增殖率(136.42±15.36)、迁移细胞数(117.14±15.92)、Bcl-2(1.56±0.20)和beclin-1(0.96±0.13)、LC3Ⅱ(1.86±0.32)和collagenⅠ(0.82±0.09)升高,KFs细胞凋亡率(2.63±0.32)、Bax(0.23±0.04)和Caspase-3(0.19±0.03)降低(P<0.05);miR-203a-5p模拟组则相反。结论miR-203a-5p通过Bcl-2/beclin-1通路能抑制KFs细胞增殖和迁移,促进KFs细胞凋亡。

Objective To investigate the effect of miR-203a-5p on the proliferation,migration and apoptosis of keloid fibroblasts.Methods From January to December 2020,the miRNAs targeting Bcl-2 were screened and transfected into KFS cells.The cell proliferation,migration and apoptosis were detected.At the same time,the protein expression levels of Bcl-2,Bax,Caspase-3,beclin-1,LC3Ⅱand collagenⅠwere detected.Results miR-203a-5p had potential binding sites with Bcl-2;miR-203a-5p mRNA in keloids was lower than that in normal skin,and Bcl-2 mRNA was higher than that in normal skin(P<0.05).KFs cell proliferation rate(136.42±15.36),number of migrating cells(117.14±15.92),Bcl-2(1.56±0.20)and beclin-1(0.96±0.13),LC3Ⅱ(1.86±0.32)and collagenⅠ(0.82±0.09)in miR-203a-5p inhibitor group increased,KFs cell apoptosis rate(2.63±0.32),Bax(0.23±0.04)and Caspase-3(0.19±0.03)decreased(P<0.05).The effect in miR-203a-5p mimic group was opposite.Conclusion miR-203a-5p can inhibit the proliferation and migration of KFs cells through the Bcl-2/beclin-1 pathway,and promote the apoptosis of KFs cells.