F10基因过表达及沉默对绒癌细胞系JAR侵袭相关蛋白酶表达的影响  

Effects of the F10 gene overexpression and silence on the expressions of invasion-related proteinases in choriocarcinoma cell line JAR

作　　者：苏晓华[1] 庞战军[1] 

机构地区：[1]南方医科大学南方医院妇产科,广州医学硕士研究生510515

出　　处：《医学研究生学报》2015年第4期350-354,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81170595)

摘　　要：目的 F10基因是本研究前期发现在葡萄胎中高表达的新基因,本文为明确F10过表达和沉默对绒癌细胞系JAR侵袭性的影响,探讨F10基因与部分侵袭相关蛋白的关系。方法通过细胞转染技术及RNA干扰技术,分别建立和筛选出F10基因稳定过表达及沉默的绒癌细胞系JAR。取SPF级裸鼠30只,随机均分为F10过表达组、未处理组、F10沉默组,每组10只,依次接种F10基因过表达的JAR细胞、未处理的JAR细胞和F10基因沉默的JAR细胞株,于接种5周后处死裸鼠收获皮下肿瘤,采用免疫组织化学及Western blot检测方法,比较不同组瘤体组织中基质金属蛋白酶(matrix metalloproteinase,MMPs)及金属蛋白酶组织抑制物-1(tissue inhibitors of metalloproteinases-1,TIMP-1),血浆纤溶酶原激活物抑制因子(plasminogen activator inhibitor-1,PAI-1)等侵袭相关蛋白酶的表达差异。结果免疫组化法检测结果显示:F10、MMP-2、8、11、16、19在F10过表达组、未处理组、F10沉默组中间的表达依次递减,差异有统计学意义(P<0.05),TIMP-1、PAI-1在F10过表达组中分别为0.118±0.029、0.174±0.005,在未处理组中分别为0.214±0.028、0.289±0.009,在F10沉默组中分别为0.430±0.017、0.517±0.074,3组间的表达差异有统计学意义(P<0.05)。Western blot结果显示,F10、MMP-2、8、11、16、19蛋白在F10沉默组、未处理组、F10过表达组依次递增(P<0.001),而TIMP-1及PAI-1蛋白表达水平依次递减(P<0.001)。结论 F10通过上调MMP-2、8、11、16、19的表达,下调TIMP-1、PAI-1表达,参与调节绒癌细胞的增殖,从而可能参与调节绒癌的侵袭和转移。Objective The F10 gene was found in the initial stage of our study to be highly expressed in hydatidiform mole. The aim of this study was to investigate the effects of theoverexpression or depletion of F10 on the invasiveness of the choriocarcinoma cell line JAR and explore the relationship of F10 expression with the invasiveness and metastasis ofchoriocarcinoma. Methods Using cell transfection and RNA interference technology, we established JAR choriocarcinoma cell lines with stablyoverexpressedor silenced F10 gene.We randomly and equally assigned 30 SPF nude mice into the three groups to receive the injection of JAR cellswith overex-pressed F10 （ F10 overexpression group） , untreated JAR cells （ control group） , and JAR cells with silenced F10（ F10 silence group） . At 5 weeks after the JAR cell injection, we harvested the subcutaneous tumor tissues from the mice, determined the expressions of ma-trix metalloproteinases （MMP）, tissue inhibitors of metalloproteinase-1（TIMP-1）, and plasminogen activator inhibitor-1（PAI-1）, and compared by Western blot and immunohistochemistry, and compared the expressions among the three groups of mice. Results Im-munohistochemistryshowed significantlyup-regulated expressions of MMP-2, -8, -11, -16, and-19 and down-regulated expressions of TIMP-1 and PAI-1 in the subcutaneous tumor tissues in the F10 overexpression group as compared with the control and F10 silence groups （ P〈0.05） .Western blot also exhibitedextremely significantly up-regulated expressions of MMP-2,-8,-11,-16, and -19 pro-teins anddown-regulated expressions of TIMP-1 and PAI-1 proteins in the F10 overexpression groupin comparison with the control and F10 silence groups（ P〈0.01 ） . Conclusion By up-regulating the expressions of MMPs and down-regulating the expressions of TIMP-1 and PAI-1, the F10 gene might be an upstream stimulating factor involved in the proliferation, invasiveness, and metastasis of-choriocarcinoma cells.

关 键 词：绒癌 F10基因 基质金属蛋白酶 血浆纤溶酶原激活物抑制因子 JAR细胞 

分 类 号：R737.33[医药卫生—肿瘤]