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长链非编码 RNA SNHG14 通过调控 miR-5590-3p/YY1 轴促进前列腺癌的恶性转化。

Long noncoding RNA SNHG14 promotes malignancy of prostate cancer by regulating with miR-5590-3p/YY1 axis.

机构信息

The First School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 May;24(9):4697-4709. doi: 10.26355/eurrev_202005_21158.

Abstract

OBJECTIVE

Studies have demonstrated that long non-coding RNAs (lncRNAs) are important in the development and prognosis of prostate cancer. The aim of this study was to investigate the functions and mechanism of lnc-SNHG14 in prostate cancer.

PATIENTS AND METHODS

Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) or Western blot (WB) were performed to detect mRNA expressions of SNHG14 and miR-5590-3p, and the protein levels of Yin Yang-1 (YY1) in prostate cancer tissues, adjacent tissues, and cancer cell lines. The correlation analysis was used to analyze the correlations between SNHG14, miR-5590-3p, and YY1. Kaplan-Meier survival analysis was used to analyze the overall survival for prostate cancer patients. Cell Counting Kit-8 (CCK-8) assay was performed to measure cell proliferation ability and flow cytometry assay was used to detect cell apoptotic rate. Besides, transwell assay was used to measure cell invasion ability. In addition, WB was performed to measure protein expressions in prostate cancer cell lines. Finally, Luciferase reporter assay was performed to verify the binding sites between SNHG14 and miR-5590-3p, miR-5590-3p, and YY1.

RESULTS

The results showed that SNHG14 was significantly increased in prostate cancer tissues and prostate cancer cell lines, which were related with advanced stage and poor diagnosis for prostate cancer patients. MiR-5590-3p was reduced in prostate cancer tissues and cell lines, which were negatively correlated with SNHG14. YY1 was found to be increased in prostate cancer tissues, which was negatively correlated with miR-5590-3p and positively correlated with SNHG14. Furthermore, SNHG14 knockdown inhibited cell proliferation, invasion, and promoted cell apoptosis in DU145 cells. In addition, protein expressions of Cyclin D1, Bcl-2, and N-cadherin were repressed, and the levels of Bax, Cleaved Caspase-3, and E-cadherin were increased. Besides, miR-5590-3p inhibition promoted cell proliferation and invasion, and inhibited apoptosis in DU145 cells. Importantly, Luciferase reporter assay proved that SNHG14 could directly sponge with miR-5590-3p, which could bind with YY1 and regulate the functions of cancer cell. Finally, we proved that SNHG14 regulated cell proliferation, cell apoptosis, and invasion via miR-5590-3p/ YY1 axis in prostate cancer.

CONCLUSIONS

Above all, we found that SNHG14 was increased in prostate cancer patients, which was related with future diagnosis for prostate cancer patients. Of note, we discovered that SNHG14 could promote cell proliferation, invasion, and repress cell apoptosis via miR-5590-3p/YY1 axis in prostate cancer, which might provide a new target for treating prostate cancer.

摘要

目的

研究表明长链非编码 RNA(lncRNAs)在前列腺癌的发生和预后中起重要作用。本研究旨在探讨 lnc-SNHG14 在前列腺癌中的功能和机制。

患者与方法

采用实时定量聚合酶链反应(qRT-PCR)或 Western blot(WB)检测前列腺癌组织、相邻组织和癌细胞系中 SNHG14 和 miR-5590-3p 的 mRNA 表达水平,以及 Yin Yang-1(YY1)的蛋白水平。采用相关性分析分析 SNHG14、miR-5590-3p 和 YY1 之间的相关性。Kaplan-Meier 生存分析用于分析前列腺癌患者的总生存率。细胞计数试剂盒-8(CCK-8)检测用于测量细胞增殖能力,流式细胞术检测用于检测细胞凋亡率。此外,Transwell 检测用于测量细胞侵袭能力。此外,还进行了 WB 以测量前列腺癌细胞系中的蛋白表达。最后,进行了荧光素酶报告基因检测以验证 SNHG14 与 miR-5590-3p、miR-5590-3p 与 YY1 之间的结合位点。

结果

结果表明,SNHG14 在前列腺癌组织和前列腺癌细胞系中显著升高,与前列腺癌患者的晚期和预后不良有关。miR-5590-3p 在前列腺癌组织和细胞系中减少,与 SNHG14 呈负相关。YY1 被发现存在于前列腺癌组织中,与 miR-5590-3p 呈负相关,与 SNHG14 呈正相关。此外,SNHG14 敲低抑制了 DU145 细胞的增殖、侵袭,并促进了细胞凋亡。此外,细胞周期蛋白 D1、Bcl-2 和 N-钙黏蛋白的表达受到抑制,Bax、Cleaved Caspase-3 和 E-钙黏蛋白的水平增加。此外,miR-5590-3p 抑制促进了 DU145 细胞的增殖和侵袭,并抑制了细胞凋亡。重要的是,荧光素酶报告基因检测证实 SNHG14 可以直接与 miR-5590-3p 结合,从而与 YY1 结合并调节癌细胞的功能。最后,我们证明 SNHG14 通过 miR-5590-3p/YY1 轴在前列腺癌中调节细胞增殖、细胞凋亡和侵袭。

结论

综上所述,我们发现前列腺癌患者 SNHG14 升高,与前列腺癌患者的未来诊断有关。值得注意的是,我们发现 SNHG14 可以通过 miR-5590-3p/YY1 轴促进前列腺癌细胞的增殖、侵袭和抑制细胞凋亡,这可能为治疗前列腺癌提供新的靶点。

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