siRNA干扰<em>MMP-9</em>和<em>FAK</em>双基因抑制小鼠黑色素瘤生长和在体迁移

doi:10.13523/j.cb.20160505

中国生物工程杂志

2016, Vol. 36

Issue (5): 34-39 DOI: 10.13523/j.cb.20160505

研究报告

siRNA干扰MMP-9和FAK双基因抑制小鼠黑色素瘤生长和在体迁移

胡娜¹, 刘清¹, 唐照勇¹, 汤禾静¹, 敖澜¹, 赵紫豪¹, 方廖琼^1,2

1. 西南大学生物技术学院重庆 400716;
2. 重庆医科大学生物医学工程学院省部共建超声医学工程国家重点实验室超声医学工程重庆市市级重点实验室重庆 400016

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-qiong^1,2

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

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摘要：

目的:观察干扰MMP-9和FAK双基因对恶性黑色素瘤高转移细胞B16F10体内转移的影响。方法:构建PGV102-MMP9-siRNA、PGV102-FAK-siRNA重组质粒载体,脂质体^TM2000介导转染小鼠黑色素瘤B16F10细胞,RT-PCR检测基因的干扰效果;建立C57BL/6小鼠皮下移植瘤模型观察细胞在体成瘤和肿瘤的生长情况,常规组织切片,H&E染色观察肿瘤组织病理学特征;经C57BL/6小鼠尾静脉注射细胞5×10⁵个/只,24天后计数小鼠肺转移结节数评价肿瘤细胞在体迁移能力。结果:RT-PCR结果表明,重组质粒转染细胞组的MMP-9和FAK的mRNA水平显著低于正常细胞组(P<0.01),转染细胞组C57BL/6小鼠皮下成瘤的肿瘤生长速率、黑色素瘤肺转移结节数明显低于正常细胞组(P<0.01)。结论:干扰B16F10细胞MMP-9和FAK双基因可明显抑制小鼠体内恶性肿瘤的生长和迁移。

关键词： 转移; MMP-9 FAK; siRNA干扰

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.

Key words: siRNA interference MMP-9 FAK Migration

收稿日期: 2015-11-30 出版日期: 2016-05-25

ZTFLH:

Q789

基金资助:

国家"973"计划资助项目(2012CB722402)

通讯作者: 方廖琼 E-mail: lqfang06@163.com

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引用本文:

胡娜, 刘清, 唐照勇, 汤禾静, 敖澜, 赵紫豪, 方廖琼. siRNA干扰MMP-9和FAK双基因抑制小鼠黑色素瘤生长和在体迁移[J]. 中国生物工程杂志, 2016, 36(5): 34-39.

HU Na, LIU Qing, TANG Zhao-yong, TANG He-jing, AO Lan, ZHAO Zi-hao, FANG Liao-qiong. siRNA Inhibits the Growth and Migration of Mouse Melanoma by MMP-9 and FAK Gene. China Biotechnology, 2016, 36(5): 34-39.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160505 或 https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I5/34

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