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miR-495通过抑制黑色素瘤细胞中的PBX3来抑制细胞增殖、迁移和侵袭,并诱导细胞凋亡。

miR-495 inhibits proliferation, migration, and invasion and induces apoptosis via inhibiting PBX3 in melanoma cells.

作者信息

Chen Guangxiong, Xie Yijie

机构信息

Department of Dermatology, Yinzhou People's Hospital, Ningbo City, Zhejiang Province, China.

出版信息

Onco Targets Ther. 2018 Apr 5;11:1909-1920. doi: 10.2147/OTT.S152362. eCollection 2018.

Abstract

BACKGROUND

Amounting evidence indicate that miRNAs play an important role in the development of various cancers. MiR-495 is a potential tumor suppressor in cancers, however its role in melanoma is still elusive. The study aimed to investigate the role of miR-495 and the underlying mechanisms in melanoma cells.

METHODS

The levels of miR-495 in melanoma tissues and cell lines were measured by quantitative real-time polymerase chain reaction. Mimics of miR-495 was transfected into human melanoma cells A375 and MeWo. Cell viability of miR-495-transfected cells was assayed by MTT assay. Cell migration and invasion of miR-495 transfected cells were measured by wound healing assay and transwell assay, respectively. Nucleosome enzyme-linked immunosorbent assay was performed to measure the apoptosis induced by overexpression of miR-495. Luciferase reporter assays were performed to verify the interaction between miR-495 and its target PBX3.

RESULTS

It was found that the expression levels of miR-495 were down-regulated in melanoma tissues and cells. Moreover, overexpression of miR-495 inhibited melanoma cell proliferation, migration and invasion in vitro. PBX3 was identified as a target for inhibition by miR-495 and was confirmed by luciferase assay, quantitative real-time polymerase chain reaction and western blot. We also indicated that silencing of PBX3 also repressed melanoma cell proliferation, migration and invasion in vitro.

CONCLUSION

In summary, our findings demonstrated that miR-495 functions as a tumor suppressor in human melanoma via directly targeting PBX3.

摘要

背景

越来越多的证据表明,微小RNA(miRNA)在多种癌症的发展中发挥着重要作用。miR-495在癌症中是一种潜在的肿瘤抑制因子,然而其在黑色素瘤中的作用仍不清楚。本研究旨在探讨miR-495在黑色素瘤细胞中的作用及其潜在机制。

方法

采用定量实时聚合酶链反应检测黑色素瘤组织和细胞系中miR-495的水平。将miR-495模拟物转染到人黑色素瘤细胞A375和MeWo中。通过MTT法检测转染miR-495细胞的活力。分别通过伤口愈合试验和Transwell试验检测转染miR-495细胞的迁移和侵袭能力。进行核小体酶联免疫吸附试验以检测miR-495过表达诱导的细胞凋亡。进行荧光素酶报告基因试验以验证miR-495与其靶标PBX3之间的相互作用。

结果

发现黑色素瘤组织和细胞中miR-495的表达水平下调。此外,miR-495的过表达在体外抑制了黑色素瘤细胞的增殖、迁移和侵袭。PBX3被确定为miR-495的抑制靶点,并通过荧光素酶试验、定量实时聚合酶链反应和蛋白质印迹法得到证实。我们还表明,沉默PBX3也能在体外抑制黑色素瘤细胞的增殖、迁移和侵袭。

结论

总之,我们的研究结果表明,miR-495通过直接靶向PBX3在人类黑色素瘤中发挥肿瘤抑制作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0bb/5896685/6619905352bd/ott-11-1909Fig1.jpg

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