Yin Pin, Peng Rui, Peng Huimin, Yao Li, Sun Yan, Wen Li, Wu Tianhui, Zhou Ji, Zhang Zheng
Molecular Medicine and Cancer Research Center, Chongqing Medical University, No.1, Medical College Road, Chongqing, 400016, China.
Mol Biotechnol. 2015 Jan;57(1):1-11. doi: 10.1007/s12033-014-9796-3.
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.
最近的研究表明，炎症与肺癌有关。然而，导致致癌作用的肺部炎症的确切原因尚不清楚。微小RNA（miRNA）是一组内源性非编码小RNA，在许多疾病中通过炎症反应调节靶向mRNA的活性。据报道，miR-451与肺癌的发展和胶质瘤的转移有关。但miR-451对肺癌细胞增殖、迁移和侵袭的影响尚不清楚。为了探讨肺癌发生发展的分子机制，我们研究了人miR-451对肺癌细胞系A549增殖、侵袭和转移的影响。将miR-451表达构建体构建到pGenesil-1.1中并转染到A549细胞中。结果表明，重组载体经测序验证。通过实时RT-PCR在A549中过表达miR-451。此外，通过MTT法、Transwell侵袭试验和伤口愈合试验，与对照组和A549组相比，miR-451组细胞的增殖、侵袭和转移受到显著抑制。通过蛋白质印迹法检测发现，miR-451组肺癌转移因子（MMP-2、MMP-9、VEGF和CXCR4）减少。此外，通过生物信息学分析和双荧光素酶报告基因试验证明，炎症相关基因PSMB8是miR-451的靶标。与对照组和A549组相比，miR-451组PSMB8和NOS2的蛋白表达降低。因此，我们的研究结果表明，与PSMB8/NOS2炎症因子相关的miR-451可能抑制肺癌的发展和迁移，为miR-451在肺癌中的作用提供了证据。
