miR-181a 通过靶向 CDK1 抑制非小细胞肺癌细胞增殖。

MiR-181a inhibits non-small cell lung cancer cell proliferation by targeting CDK1.

机构信息

Department of Oncology, Fuzhou Pulmonary Hospital of Fujian, Fuzhou, Fujian, China.

Infectious Disease Department, The People's Hospital of Ningde City, Ningde, Fujian, China.

出版信息

Cancer Biomark. 2017 Dec 6;20(4):539-546. doi: 10.3233/CBM-170350.

DOI:10.3233/CBM-170350

PMID:28946554

Abstract

BACKGROUND

MicroRNAs (miRNAs) emerge as important regulators involved in malignant progression in some tumors. MiR-181a has been found to function as a tumor suppressor in some tumors including non-small cell lung cancer (NSCLC). However, the functional role of miR-181a in NSCLC still needed to be investigated.

METHODS

The expression of miR-181a were determined by qRT-PCR, the association between miR-181a and clinicopathological data were performed by chi-square test and survival analysis were evaluated by Kaplan-Meier curve and log rank test. Cell proliferation and invasion were assessed by CCK8, cell colony formation and transwell assays. Luciferase reporter assay demonstrated that CDK1 was a target of miR-181a. Western blot assay detected the relative protein expression.

RESULTS

In the study, our results showed that miR-181a was significantly down-regulated in non-small cell lung cancer (NSCLC) tissues and cell lines. MiR-181 expression levels were significantly associated with histological grade, N status and TNM stage in the patients and lower miR-181a predicted a poor prognosis in NSCLC patients. Furthermore, upregulation of miR-181a significantly suppressed the NSCLC cell proliferation, colony formation, and cell invasion capacities. Moreover, upregulation of miR-181a inhibited CyclinB1 and CyclinD1 expression in NSCLC cells. Luciferase activity assay results demonstrated CDK1 was a direct target of miR-181a and miR-181a inhibited cell proliferation by regulating the mRNA and protein levels of CDK1 in NSCLC cells.

CONCLUSION

These data suggested that miR-181a plays a tumor suppressor and may be a potential therapeutic target for NSCLC patients.

摘要

背景

微小 RNA（miRNAs）作为一些肿瘤中恶性进展的重要调节因子而出现。miR-181a 在一些肿瘤中作为肿瘤抑制因子发挥作用，包括非小细胞肺癌（NSCLC）。然而，miR-181a 在 NSCLC 中的功能作用仍需进一步研究。

方法

通过 qRT-PCR 确定 miR-181a 的表达，通过卡方检验和生存分析评估 miR-181a 与临床病理数据的相关性，通过 Kaplan-Meier 曲线和对数秩检验评估生存分析。通过 CCK8、细胞集落形成和 Transwell 测定评估细胞增殖和侵袭。荧光素酶报告测定显示 CDK1 是 miR-181a 的靶标。Western blot 测定检测相对蛋白表达。

结果

在本研究中，我们的结果表明 miR-181a 在非小细胞肺癌（NSCLC）组织和细胞系中显著下调。miR-181 表达水平与患者的组织学分级、N 状态和 TNM 分期显著相关，miR-181a 低表达预示 NSCLC 患者预后不良。此外，miR-181a 的上调显著抑制 NSCLC 细胞的增殖、集落形成和细胞侵袭能力。此外，miR-181a 的上调抑制了 NSCLC 细胞中 CyclinB1 和 CyclinD1 的表达。荧光素酶活性测定结果表明 CDK1 是 miR-181a 的直接靶标，miR-181a 通过调节 NSCLC 细胞中 CDK1 的 mRNA 和蛋白水平抑制细胞增殖。