| miR-147a靶向INHBA-TGF-β1/Smads信号通路对宫颈癌HeLa细胞增殖、迁移与侵袭的作用 |
| 投稿时间：2025-01-21 修订日期：2025-02-25 点此下载全文 |
| 引用本文：张春瑜,邹俊凯,周琦.miR-147a靶向INHBA-TGF-β1/Smads信号通路对宫颈癌HeLa细胞增殖、迁移与侵袭的作用[J].医学研究杂志,2025,54(8):80-87 |
| DOI：
10.11969/j.issn.1673-548X.2025.08.014 |
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| 基金项目:新疆维吾尔自治区自然科学基金资助项目(2022D01C805) |
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| 中文摘要:目的 探讨miR-147a靶向抑制素亚基βA(inhibin subunit beta A,INHBA)介导转化生长因子-β1(transforming growth factor-β1,TGF-β1)/Smads信号通路在宫颈癌HeLa细胞侵袭、转移过程中的作用。方法 将miR-147a模拟物、miR-147a抑制剂及其阴性对照(mimics NC和inhibitor NC)转染至HeLa细胞中,分为5组,即control组(空白对照)、mimics NC组、miR-147a mimics组、inhibitor NC组和miR-147a inhibitor组。采用MTT法检测miR-147a干预后HeLa细胞的增殖活性,采用Transwell和细胞划痕实验检测miR-147a干预后HeLa细胞的侵袭转移能力,采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction, RT-qPCR)法检测不同分组中miR-147a与INHBA的mRNA表达变化,采用双荧光素酶报告基因检测miR-147a与靶基因INHBA之间的关系,采用Western blot法检测不同分组中TGF-β1、TGFBR1、Smad3、Smad2、INHBA的蛋白表达情况,采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测TGF-β1的表达变化。结果 与空白对照及阴性对照比较,miR-147a mimics抑制细胞增殖、降低细胞侵袭和迁移能力、降低细胞愈合能力,miR-147a inhibitor促进细胞增殖、增加侵袭和迁移能力、提高细胞愈合能力；miR-147a mimics组的miR-147a显著升高,靶基因INHBA显著降低(P<0.05)；miR-147a inhibitor组的miR-147a显著降低,靶基因INHBA显著升高(P<0.05)；miR-147a mimics+WT质粒组的荧光强度比值较mimics NC+WT组显著降低(P<0.05)；miR-147a mimics组的TGF-β1、TGFBR1、Smad3、Smad2、INHBA的表达水平显著降低(P<0.05)；miR-147a inhibitor组的TGF-β1、TGFBR1、Smad3、Smad2、INHBA的表达水平显著升高(P<0.05)。结论 miR-147a靶向INHBA介导TGF-β1/Smads信号通路,在抑制宫颈癌HeLa细胞增殖、侵袭和迁移过程中发挥重要作用,为宫颈癌的治疗提供了潜在的分子靶点和理论依据。 |
| 中文关键词:miR-147a INHBA TGF-β1/Smads 宫颈癌 |
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| Effect of miR-147a Targeting INHBA-TGF-β1/Smads Signaling Pathway on Proliferation, Migration and Invasion of Cervical Cancer HeLa Cells. |
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| Abstract:Objective To investigate the role of miR-147a targeting inhibin subunit beta A (INHBA) in transforming growth factor-β1 (TGF-β1)/Smads signaling pathway in the invasion and metastasis of cervical cancer HeLa cells. Methods The miR-147a mimics, miR-147a inhibitors and their negative controls (mimicsNC and inhibitor NC) were transfected into HeLa cells and divided into five groups:control group (blank control), mimics NC group, miR-147a mimics group, inhibitor NC group, and miR-147a inhibitor group. The proliferation activity of HeLa cells after miR-147a intervention was detected by the MTT method, the invasion and metastasis abilities of HeLa cells after the intervention of miR-147a were detected by Transwell and cell scratch assay, the mRNA expression changes of miR-147a and INHBA in different groups were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), the relationship between miR-147a and the target gene INHBA was detected by dual-luciferase reporter gene assay, the protein expressions of TGF-β1, TGFBR1, Smad3, Smad2 and INHBA in different groups were detected by Western blot, and the expression changes of TGF-β1 were detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with blank control and negative control, miR-147a mimics inhibited cell proliferation, decreased cell invasion and migration ability, and decreased cell healing ability. miR-147a inhibitor promoted cell proliferation, increased invasion and migration ability, and improved cell healing ability. In the miR-147a mimics group, miR-147a was significantly increased, and the target gene INHBA was significantly decreased (P<0.05). In the miR-147a inhibitor group, miR-147a was significantly decreased, and the target gene INHBA was significantly increased (P<0.05). The fluorescence intensity ratio of miR-147a mimics+WT group was significantly lower than that of mimics NC+WT group (P<0.05). The expression levels of TGFBR1, TGF-β1, Smad3, Smad2 and INHBA in miR-147a mimics group were significantly decreased (P<0.05). The expression levels of TGFBR1, TGF-β1, Smad3, Smad2, and INHBA in miR-147a inhibitor group were significantly increased (P<0.05). Conclusion miR-147a targets the TGF-β1/Smads signaling pathway mediated by INHBA, which plays an important role in inhibiting the proliferation, invasion and migration of cervical cancer HeLa cells, and provides a potential molecular target and theoretical basis for the treatment of cervical cancer. |
| keywords:miR-147a INHBA TGF-β1/Smads Cervical cancer |
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