| 高栋梁,张雷,张宏峰,等.微小 RNA-4463对人瘢痕疙瘩成纤维细胞增殖和侵袭的影响[J].安徽医药,2021,25(6):1140-1143. |
| 微小 RNA-4463对人瘢痕疙瘩成纤维细胞增殖和侵袭的影响 |
| Effect of miR-4463 on proliferation and invasion of human keloid fibroblasts |
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| DOI：10.3969/j.issn.1009-6469.2021.06.019 |
| 中文关键词: 瘢痕疙瘩 成纤维细胞 miR-4463 Bcl2相关致病基因 4(BAG4) |
| 英文关键词: Keloid Fibroblasts MiR-4463 Bcl2 Associated Athanogene 4 (BAG4) |
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| 中文摘要: |
| 目的研究微小 RNA-4463(miR-4463)对人瘢痕疙瘩成纤维细胞增殖、迁移、侵袭的影响以及机制。方法收集延安大学附属医院 2017年 12月至 2019年 6月整形手术病人 40例，每例切除瘢痕疙瘩组织 1个以及邻近正常皮肤组织 1个，利用定量聚合酶链反应(qPCR)检测人正常成纤维细胞和瘢痕疙瘩成纤维细胞中 miR-4463的表达量；将 miR-4463模拟物对照质粒和 miR-4463模拟物分别在 Lipofectamine 2000介导下转染入瘢痕疙瘩成纤维细胞，分为 miR-NC组和 miR-4463组；常规培养的瘢痕疙瘩成纤维细胞作为对照； qPCR检测转染后 miR-4463的表达量； qPCR检测上调 miR-4463对瘢痕疙瘩纤维化相关基因 Col 1 A1、Col 3 A1表达的影响；细胞计数试剂盒( CCK-8)检测上调 miR-4463对瘢痕疙瘩成纤维细胞增殖的影响， Transwell实验检测细胞的迁移、侵袭；采用 TargetScan软件预测 miR-4463与 B细胞淋巴瘤 -2(Bcl-2)相关的致病基因 BAG4的靶向关系，双荧光素酶报告基因进一步进行验证。结果 miR-4463在瘢痕疙瘩成纤维细胞中的表达量较人正常成纤维细胞显著降低［(0.218± 0.036)比( 1.000±0.071)，P<0.05］；与对照组相比，转染 miR-4463模拟物明显增加瘢痕疙瘩成纤维细胞中 miR-4463的表达量［( 1.000±0.073)比( 3.313±0.242)， P<0.05］；上调 miR-4463显著抑制瘢痕疙瘩纤维化相关基因 Col 1 A1［( 1.000±0.065)比 |
| 英文摘要: |
| Objective To study the effects of microRNA-4463 (miR-4463) on the proliferation, migration and invasion of human keloid fibroblasts and its mechanism.Methods Forty patients with plastic surgery in Yan′an University Affiliated Hospital from December 2017 to June 2019 were collected. One keloid tissue and one adjacent normal skin tissue were excised from each patient. The expression of mir-4463 in human normal fibroblasts and keloid fibroblasts was detected by quantitative polymerase chain reaction (qP‐CR). The control plasmid and the mimic of miR-4463 were respectively transfected into keloid fibroblasts by Lipofectamine 2000 and assigned into miR-NC group and miR-4463 group, while the conventional cultured keloid fibroblasts were selected as controls. The expression of miR-4463 after transfection was detected by qPCR. qPCR was used to detect the effect of up-regulated miR-4463 on the expression of keloid fibrosis related genes Col 1 A1 and Col 3 A1. Cell counting kit-8 (CCK-8) was used to detect the effect of up-regulation of miR-4463 on the proliferation of keloid fibroblasts. The cell migration and invasion were analyzed by Transwell assay. The targeting relationship between miR-4463 and Bcl-2-associated athanogene (BAG4) was predicted by TargetScan and further validated by double luciferase reporter gene.Results The expression of miR-4463 in keloid fibroblasts was significantly lower than that in human normal fibroblasts [(0.218±0.036) vs. (1.000±0.071), P<0.05]. Compared with the control group, the expression of miR-4463 in keloid fibroblasts transfection with miR-4463 mimics was significantly increased [(1.000±0.073) vs. (3.313±0.242), (P<0.05)]. Up-regulation of miR-4463 significantly inhibited the expression of Col 1 A1 [(1.000±0.065) vs. (0.334±0.010)] and Col 3 A1 [(1.000±0.070) vs. (0.301± 0.008)] in keloid fibrosis (P<0.05). Up-regulation of miR-4463 inhibited the proliferation of keloid fibroblasts [(0.836±0.081) vs. (0.656±0.072), P<0.05], and decreased the number of migrating [(153.269±12.471) vs. (82.363±7.254)] and invading [(73.623±6.451) vs. (36.459±3.235)] cells (P<0.05). BAG4 was the downstream target gene of miR-4463.Conclusion miR-4463 may inhibit the expression of fibrosis-related collagen genes in extracellular matrix by regulating BAG4, and inhibit the proliferation, migration and invasion of keloid fibroblasts. |
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