|
|
|
| siRNA Inhibits the Growth and Migration of Mouse Melanoma by MMP-9 and FAK Gene |
| HU Na1, LIU Qing1, TANG Zhao-yong1, TANG He-jing1, AO Lan1, ZHAO Zi-hao1, FANG Liao-qiong1,2 |
1. College of Biotechnology, Southwest University, Chongqing 400716, China;
2. College of Biomedical Engineering, Chongqing Medical University State Key Laboratory of Ultrasound Engineering in Medicine Co-founded by Chongqing and the Ministry of Science, Chongqing 400016, China |
|
|
|
Abstract Objective:To observe the effect of combined-silencing MMP-9 and FAK on migration of malignant melanoma highly metastatic B16F10 cells in vivo. Methods:Construct recombinant plasmid vectors pGV102-MMP9-siRNA and pGV102-FAK-siRNA, then Lipofectamine TM2000 mediated the plasmid vectors were transfected into B16F10 cells. To observe the growth of the tumor, the C57BL/6 mouse model of subcutaneous transplantation was established, and the pathological characteristics of tumor tissue was observed by H&E staining. B16F10 cells were stably intravenously injected into C57BL/6 mice by 5×105. After 24 days of injection, the number of mouse lung metastasis nodules was counted to evaluate the migration ability of tumor cells in vivo. Results:RT-PCR results showed that the mRNA level of MMP-9 and FAK in the transfected cells was significantly reduced than that of normal cells(P<0.01). Compared with the normal cells, the tumor growth rate and the number of lung metastasis nodules of C57BL/6 mice in the transfected cells were significantly reduced(P<0.01). Conclusion:Interference MMP-9 and FAK can significantly inhibit malignant tumor growth and metastasis in vivo.
|
|
Received: 30 November 2015
Published: 25 May 2016
|
|
|
|
|
| [1] 朱耀德. 恶性黑色素瘤. 徐州医学院学报, 1992, 1: 81-87. Zhu Y D. Malignant melanoma. Acta Academic Medicinae Xuzhou, 1982, 1:81-87.
[2] 简雪平, 文辉才,韩超,等. 恶性黑色素瘤的基因治疗新进展. 南昌大学学报, 2013, 53(1): 90-92. Jian X P, Wen H C, Han C, et al. The progress in gene therapy of malignant melanoma. Journal of Nanchang University, 2013, 53(1): 90-92.
[3] Qian H, Yu J, Li Y, et al. RNA interference of metastasis-associated gene inhibits metastasis of B16F10 melanoma cells in a C57BL/6 mouse model. Biol Cell, 2007, 99(10): 573-581.
[4] Li S, Dong W, Zong Y, et al. Polyethylenimine-complexed plasmid particles targeting focal adhesion kinase function as melanoma tumor therapeutics. Mol Ther, 2007, 15(3):515-523.
[5] Terao S, Shirakawa T, Goda K, et al. Recombinant interleukin-2 enhanced the antitumor effect of ADV/RSV-HSV-tk/ACV therapy in a murine bladder cancer model. Anticancer Res, 2005, 25(4): 2757-2760.
[6] Huang Q, Pu P, Xia Z, et al. Exogenous wt-p53 enhances the antitumor effect of HSV-TK/GCV on C6 glioma cells. J Neurooncol, 2007, 82(3):239-248.
[7] 汤禾静,唐照勇,刘隆兴,等. siRNA联合沉默MMP-9和FAK基因对小鼠黑色素瘤高转移细胞B16F10体外侵袭和迁移的影响. 中国生物工程杂志, 2014, 34(9): 40-47. Tang H J, Tang Z Y, Liu L X, et al. Effect of siRNA combined-silencing MMP-9 and FAKon invasion and migration of mouse melanoma highly metastatic cells B16F10in vitro. China Biotechnology, 2014, 34(9): 40-47.
[8] Yuan J, Dutton C M, Scμlly S P. RNAi mediated MMP-1 silencing inhibits human chondrosarcoma invasion. J Orthop Res, 2005, 23(6):1467-1474.
[9] Han D C, Shen T L, Guan J L. Role of Grb7 targeting to focal contacts and its phosphorylation by focal adhesion kinase in regμlation of cell migration. J Biol Chem, 2000, 275(37): 28911-28917.
[10] Lakka S S, Gondi C S, Yanamandra N, et al. Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis. Oncogene, 2004, 23(27): 4681-4689.
[11] Gencer S, Cebeci A, Irmak-Yazicioglu M B. silencing of the MMP-3 gene by siRNA transfection in gastric cancer AGs cells. J Gastrointestin Liver Dis, 2011, 20(1): 19-26.
[12] 孙亚男, 刘鸣, 孙艳,等. MMP-9基因RNAi重组慢病毒的构建及其对喉癌细胞侵袭的抑制. 中国耳鼻咽喉头颈外科, 2008, 15(7): 391-393. Sun Y N, Liu M, Sun Y, et al. Construction of lentiviral vector of RNA inter ference of MMP-9 gene and its inhibition on invasion of laryngeal carcinoma cells. Head and Neck Surgery of China., 2008, 15(7): 391-393.
[13] Jiang H, Liu L, Ye J, et al. Focal adhesion kinase serves as a marker of cervical lymph node metastasis and is a potential therapeutic target in tongue cancer. J Cancer Res Clin Oncol, 2010, 136(9): 1295-1302.
[14] Hauck C R, Sieg D J, Hsia D A, et al. Inhibition of focal adhesion kinase expression or activity disrupts epidermal growth factor-stimulated signaling promoting the migration of invasive human carcinoma cells. Cancer Res, 2001, 61(19):7079-7090.
[15] Chen J S, Huang X H, Wang Q, et al. FAK is involved in invasion and metastasis of hepatocellμlar carcinoma. Clin Exp Metastasis, 2010, 27(2): 71-82.
[16] Shibata K, Kikkawa F, Nawa A, et al. Both focal adhesion kinase and c-Ras are required for the enhanced matrix metalloproteinase 9 secretion by fibronectin in ovarian cancer cells. Cancer Res, 1998, 58(5):900-903. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
