淋巴道转移舌鳞癌细胞RECK、MMP-2和MMP-9表达的动物实验研究

An animal-based experiment on RECK,MMP-2 and MMP-9 expression in lymphatic metastatic tongue carcinoma cells

目的在动物模型水平探讨RECK表达水平与MMP-2和MMP-9的关系。方法采用爪垫注射Tca8113细胞建立舌鳞癌淋巴道转移裸鼠模型,收集4周后回流淋巴结,采用免疫组化、Western blot方法检测RECK、MMP-2、MMP-9水平。结果8例舌癌淋巴结转移模型裸鼠均有MMP-2、9表达,细胞染色呈较强的阳性,而RECK未发现较强阳性细胞出现,只有可疑的阳性细胞散在出现;淋巴结转移的舌癌细胞RECK表达明显低于用于移植的Tca8113舌癌细胞(t=7.78,P<0.01);MMP-2、9表达水平均显著高于用于移植的舌癌细胞(t分别为3.06,2.25,P<0.01或0.05)。结论动物体内实验进一步证实RECK低水平表达可增加舌癌侵袭转移机率,其调控机理可能与RECK抑制MMP-2、9表达有关。

Obiective To approach the relationship between the expressive level of the reversion- inducing- cysteine - rich protein with Kazal motifs （RECK） and matrix metalloproteinases （MMP - 2 and MMP - 9） by nude mice model- based experiment. Methods The tongue carcinoma lymphatic metastasis model of the nude mouse was established by injecting Tca 8113 cells into the nail pad. We collected the regurgitant lymph nodes after 4 weeks and detected RECK, MMP - 9 and MMP - 2 levels by immunohistochemistry and Western blot assay. Results There were 8 nude mice of the tongue carcinoma lymphatic metastasis model expressing MMP- 2 and MMP- 9, and the cell staining was also strong, while there were not positive staining cells in the RECK detection, and only some scattered suspicious positive cells were found. RECK expressive level of lymphatic metastatic tongue carcinoma cell decreased significantly and were lower than the level in transplanted Tca 8113 cells （ t = 7.78, P 〈0.01）; while the expressive levels of MMP - 2 and MMP - 9 were both higher than those of the control group （ t = 3.06 and 2. 25, respectively, P 〈0.01 or 0.05）. Conclusion The animal in vivo experiments confirmed that low expression of RECK could enhance invasiveness and metastasis of tongue carcinoma, the regulating mechanism may be related to RECK inhibiting MMP- 2 and MMP- 9 expression.