Abstract
This study was aimed to evaluate the regulation mechanism of cortactin (CTTN) on matrix metalloproteinases 9 (MMP-9) and its relations with Exo70 in invasion of hepatoma carcinoma (HCC) cells. The expression levels of CTTN, Exo70 and MMP-9 were detected in normal hepatocytes and various HCC cells by real-time PCR. Then the migration and invasion ability of these cells was revealed by scratch and invasion assay. The effects of CTTN on MMP-9 and the ability of migration and invasion were evaluated by silence and overexpress CTTN. During this process, the expression of CTTN was detected by Western blot, the activity and concentration of MMP-9 in supernatant of culture medium was detected by zymography and ELISA assay. Besides, Exo70 was also inhibited to reveal its effects on MMP-9 and the migration and invasion ability of LM3. Increased expression of CTTN, MMP-9, Exo70, reduced scratch area and increased puncture cell numbers were found in HCC cells (p < 0.05). The expression of CTTN was significantly correlated with Exo70 and the migration and invasion ability of HCC (p < 0.05). In addition, the activity and concentration of MMP-9 were significantly affected by the expression level of CTTN, while the expression of MMP-9 was not influenced. Besides, Exo70-si also exhibited significantly inhibition effects on the activity and concentration of MMP-9 and puncture cell numbers (p < 0.05). A synergistic reaction may exhibited on CTTN and Exo70, which could mediate the secretion of MMPs thereby regulate tumor invasion.
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Acknowledgments
This study was supported by Scientific Research Fund of Chinese PLA Air Force General Hospital (NO. KZ2011017). The authors would like to thank Professor Xu Jin-bo, Beijing Key Laboratory of Blood Safety and Supply Technologies, for his assistance in providing open laboratory for the study.
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Zhao, G., Zhang, H., Huang, Z. et al. Cortactin and Exo70 mediated invasion of hepatoma carcinoma cells by MMP-9 secretion. Mol Biol Rep 43, 407–414 (2016). https://doi.org/10.1007/s11033-016-3972-4
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DOI: https://doi.org/10.1007/s11033-016-3972-4