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抑制 miR-9-5p 通过靶向 StarD13 抑制前列腺癌进展。

Inhibition of miR-9-5p suppresses prostate cancer progress by targeting StarD13.

机构信息

Department of Urology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, No. 26 Shengli Street, Jiang'an District, Wuhan, 430014 China.

出版信息

Cell Mol Biol Lett. 2019 Mar 8;24:20. doi: 10.1186/s11658-019-0145-1. eCollection 2019.

Abstract

BACKGROUND

This study aims to investigate the effects of inhibiting microRNA-9-5p (miR-9-5p) on the expression of StAR-related lipid transfer domain containing 13 (StarD13) and the progress of prostate cancer.

METHODS

The mRNA expression levels of miR-9-5p and StarD13 were determined in several prostate cancer cell lines. We chose DU145 and PC-3 cells for further research. The CCK8 assay was used to measure the cell viability. The cell invasion and wound-healing assays were respectively applied to evaluate invasion and migration. The expression of E-cadherin (E-cad), N-cadherin (N-cad) and vimentin were measured via western blot. DU145 and PC-3 cells overexpressing StarD13 were generated to investigate the variation in proliferation, invasion and migration. A luciferase reporter assay was used to identify the target of miR-9-5p.

RESULTS

Our results show that miR-9-5p was highly expressed and StarD13 was suppressed in prostate cancer cells. MiR-9-5p inhibition repressed the cells' viability, invasion and migration. It also increased the expression of E-cad and decreased that of N-cad and vimentin. StarD13 overexpression gave the same results as silencing of miR-9-5p: suppression of cell proliferation, invasion and migration. The bioinformatics analysis predicted StarD13 as a target gene of miR-9-5p. Quantitative RT-PCR, western blot analysis and the dual-luciferase reporter assay were employed to confirm the prediction.

CONCLUSION

Our results show that miR-9-5p plays a powerful role in the growth, invasion, migration and epithelial-mesenchymal transition (EMT) of prostate cancer cells by regulating StarD13. A therapeutic agent inhibiting miR-9-5p could act as a tumor suppressor for prostate cancer.

摘要

背景

本研究旨在探讨抑制 microRNA-9-5p (miR-9-5p) 对 StAR 相关脂质转移结构域包含蛋白 13 (StarD13) 表达和前列腺癌进展的影响。

方法

在几种前列腺癌细胞系中测定 miR-9-5p 和 StarD13 的 mRNA 表达水平。我们选择 DU145 和 PC-3 细胞进行进一步研究。CCK8 法检测细胞活力。细胞侵袭和划痕愈合试验分别用于评估侵袭和迁移。通过 Western blot 检测 E-钙黏蛋白 (E-cad)、N-钙黏蛋白 (N-cad) 和波形蛋白的表达。生成过表达 StarD13 的 DU145 和 PC-3 细胞,以研究增殖、侵袭和迁移的变化。荧光素酶报告基因检测用于鉴定 miR-9-5p 的靶基因。

结果

我们的结果表明,miR-9-5p 在前列腺癌细胞中高表达,StarD13 被抑制。miR-9-5p 抑制抑制细胞活力、侵袭和迁移。它还增加了 E-cad 的表达,降低了 N-cad 和波形蛋白的表达。StarD13 的过表达与 miR-9-5p 的沉默产生相同的结果:抑制细胞增殖、侵袭和迁移。生物信息学分析预测 StarD13 是 miR-9-5p 的靶基因。定量 RT-PCR、Western blot 分析和双荧光素酶报告基因检测证实了这一预测。

结论

我们的研究结果表明,miR-9-5p 通过调节 StarD13 在前列腺癌细胞的生长、侵袭、迁移和上皮-间充质转化 (EMT) 中发挥重要作用。抑制 miR-9-5p 的治疗剂可能作为前列腺癌的肿瘤抑制因子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1b9/6408831/7bfc129d7bed/11658_2019_145_Fig1_HTML.jpg

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