MiR-451 Suppresses Cell Proliferation and Metastasis in A549 Lung Cancer Cells
MiR-451 Suppresses Cell Proliferation and Metastasis in A549 Lung Cancer Cells
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DOI：
10.1007/s12033-014-9796-3
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发表时间：
2014
影响因子：
2.6
通讯作者：
Pin Yin;Rui Peng;Huimin Peng;Li Yao;Y. Sun;Li Wen;Tianhui Wu;Ji Zhou;Zheng Zhang
中科院分区：
文献类型：
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作者：
Pin Yin;Rui Peng;Huimin Peng;Li Yao;Y. Sun;Li Wen;Tianhui Wu;Ji Zhou;Zheng Zhang
近年来的研究表明，炎症与肺癌的发生有关。然而，肺部炎症导致癌变的确切原因尚不清楚。microRNAs（miRNAs）是一类内源性非编码小分子RNA，在多种疾病中通过炎症反应调节靶mRNA的活性。miR-451与肺癌的发生、胶质瘤的转移有关。但miR-451对肺癌细胞增殖、迁移和侵袭的影响尚不清楚。为探讨肺癌发生发展的分子机制，我们研究了人miR-451对肺癌细胞株A549增殖、侵袭和转移的影响。将miR-451表达构建体生成到pGenesil-1.1中并转染到A549细胞中。测序结果表明，重组质粒正确。实时荧光定量RT-PCR结果显示，miR-451在A549细胞中过表达。MTT法、Transwell侵袭实验和创伤愈合实验显示，与对照组和A549组相比，miR-451组细胞的增殖、侵袭和转移能力明显受到抑制。Western blot结果显示，miR-451组肺癌转移因子MMP-2、MMP-9、VEGF和CXCR 4表达降低。此外，通过生物信息学分析和双荧光素酶报告基因分析，证实炎症相关基因PSMB 8是miR-451的靶基因。与对照组和A549组相比，miR-451组PSMB 8和NOS 2的蛋白表达均降低。因此，我们的研究结果表明，与PSMB 8/NOS 2炎症因子相关的miR-451可能抑制肺癌的发生和迁移，为miR-451在肺癌中的作用提供了证据。
Recent study showed that inflammation was related to lung cancer. However, the exact cause of lung inflammation leading to carcinogenesis is unknown. MicroRNAs (miRNAs) are a group of endogenous non-coding small RNAs that regulate the activity of targeted mRNAs by inflammatory response in many diseases. MiR-451 was reported to relate to the development of lung cancer and metastasis of glioma. But the effect of miR-451 on cell proliferation, migration, and invasion of lung cancer is not really clear. In order to explore the molecular mechanism of the occurrence and development of lung cancer, we investigated the effect of human miR-451 on the proliferation, invasion, and metastasis in lung cancer cell line A549. The miR-451 expression construct was generated into pGenesil-1.1 and transfected into A549 cells. Results showed that the recombinant vectors were verified by sequencing. And miR-451 was over-expressed in A549 by real-time RT PCR. Furthermore, the proliferation, invasion, and metastasis of the cells in miR-451 group were inhibited significantly compared with those in control and A549 groups by MTT assay, Transwell invasion assay, and wound-healing assay. And the lung cancer metastasis factors (MMP-2, MMP-9, VEGF, and CXCR4) were decreased in miR-451 group by Western blot. Moreover, it was proved that inflammation-related gene-PSMB8 was a target for miR-451 by bioinformatics analysis and dual-luciferase reporter assay. And the protein expressions of PSMB8 and NOS2 were decreased in miR-451 group compared with those in control and A549 groups. Therefore, our findings indicated that miR-451 related to PSMB8/NOS2 inflammatory factors may suppress the development and migration of lung cancer, providing evidence for the role of miR-451 in lung cancer.