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微小RNA-195-5p通过靶向Yes相关蛋白1抑制宫颈癌的恶性进展。

MiR-195-5p Inhibits Malignant Progression of Cervical Cancer by Targeting YAP1.

作者信息

Liu Xiaomin, Zhou Yi, Ning Yu-E, Gu Hui, Tong Yuxin, Wang Ning

机构信息

Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang 110000, People's Republic of China.

Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital, China Medical University, Shenyang 110004, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Jan 30;13:931-944. doi: 10.2147/OTT.S227826. eCollection 2020.

Abstract

PURPOSE

Our previous studies have shown that miR-195 is reduced in cervical cancer tissues, and that upregulation of miR-195 suppressed cervical cancer cell growth and induced a cell cycle block. In this study, we aimed to further elucidate the mechanism of action between miR-195-5p and Yes-associated protein 1 (YAP1) in the malignant progression of cervical cancer.

METHODS

MiR-195-5p and YAP1 were detected using qRT-PCR in cervical cancer cells transfected with miR-195-5p mimics or inhibitor. Cell proliferation, migration, and invasion ability were detected using MTT, wound healing, and transwell invasion assays. Dual luciferase reporter assay, qRT-PCR, and Western blot analysis were used to demonstrate that YAP1 was a target of miR-195-5p.

RESULTS

Our results showed that miR-195-5p is negatively correlated with YAP1 protein levels but not with mRNA expression. Moreover, upregulation of miR-195-5p by transient transfection with miR-195-5p mimics in HeLa and SiHa cells inhibited cell proliferation, migration ability, invasiveness, and the EMT. Conversely, miR-195-5p downregulation produced opposite results. In addition, multiple miRNA target prediction sites showed that YAP1 was a potential target gene; this was confirmed by dual luciferase assay. Rescue experiments further confirmed that YAP1 is involved in miR-195-5p-mediated inhibition of proliferation, migration ability, invasiveness, and the EMT of cervical cancer cells.

CONCLUSION

Taken together, our data suggest that miR-195-5p may act as a tumor suppressor which could provide a theoretical basis for cervical cancer patient targeted therapy.

摘要

目的

我们之前的研究表明,miR-195在宫颈癌组织中表达降低,且miR-195的上调可抑制宫颈癌细胞生长并诱导细胞周期阻滞。在本研究中,我们旨在进一步阐明miR-195-5p与Yes相关蛋白1(YAP1)在宫颈癌恶性进展中的作用机制。

方法

使用qRT-PCR检测转染了miR-195-5p模拟物或抑制剂的宫颈癌细胞中miR-195-5p和YAP1的表达。采用MTT法、伤口愈合实验和Transwell侵袭实验检测细胞增殖、迁移和侵袭能力。通过双荧光素酶报告基因实验、qRT-PCR和蛋白质免疫印迹分析证明YAP1是miR-195-5p的靶标。

结果

我们的结果表明,miR-195-5p与YAP1蛋白水平呈负相关,但与mRNA表达无关。此外,在HeLa和SiHa细胞中瞬时转染miR-195-5p模拟物上调miR-195-5p可抑制细胞增殖、迁移能力、侵袭性和上皮-间质转化(EMT)。相反,miR-195-5p下调则产生相反的结果。此外,多个miRNA靶标预测位点显示YAP1是一个潜在的靶基因;双荧光素酶实验证实了这一点。挽救实验进一步证实YAP1参与了miR-195-5p介导的对宫颈癌细胞增殖、迁移能力、侵袭性和EMT的抑制作用。

结论

综上所述,我们的数据表明miR-195-5p可能作为一种肿瘤抑制因子发挥作用,这可为宫颈癌患者的靶向治疗提供理论依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dd0/6996614/3f263eba3e33/OTT-13-931-g0001.jpg

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