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

发布时间：2018-02-02 12:00

本文关键词： 绒癌 F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　出处：《医学研究生学报》2015年04期 　论文类型：期刊论文

【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(P0.05),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(P0.05)。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(P0.001),而TIMP-1及PAI-1蛋白表达水平依次递减(P0.001)。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。
[Abstract]:Objective F10 gene is a novel gene which was found to be highly expressed in hydatidiform mole. The purpose of this study is to investigate the effect of overexpression and silencing of F10 gene on the invasion of choriocarcinoma cell line JAR. To investigate the relationship between F10 gene and some invasive-associated proteins. Methods Cell transfection technique and RNA interference technique were used. A stable and silenced choriocarcinoma cell line Jar was established and screened. Thirty nude mice with SPF grade were randomly divided into F10 overexpression group and untreated F10 silencing group. In each group, 10 JAR cells were inoculated with F10 gene overexpression, untreated JAR cells and JAR cell lines silenced by F10 gene in turn. The nude mice were killed to harvest subcutaneous tumor 5 weeks after inoculation. The matrix metalloproteinase of matrix metalloproteinases in different tumor tissues were compared by immunohistochemistry and Western blot assay. MMPs) and metalloproteinase-1 tissue inhibitors of metalloproteinases-1 (TIMP-1). Plasminogen activator inhibitor-1. Results Immunohistochemical assay showed that the expression of 1% F10MP-2MMP ~ (2 +) in the F10 overexpression group was higher than that in the untreated group (P < 0.05), and the expression of PAI-1 was higher than that in the control group (P < 0.05). The middle expression of F10 silencing group decreased in turn, and the difference was statistically significant (P 0.05). The expression of TIMP-1 PAI-1 in F10 overexpression group was 0.118 卤0.029, respectively. 0.174 卤0.005, 0.214 卤0.028, 0.289 卤0.009 in untreated group and 0.430 卤0.017 in F10 silencing group, respectively. There was significant difference in the expression of MMP-2 between the three groups (P 0.05). Western blot showed that the expression of MMP-2 was 81116. 19 protein in F10 silencing group, untreated group, F10 overexpression group increased in turn (P 0.001). However, the expression of TIMP-1 and PAI-1 decreased in turn (P 0.001). Conclusion F10 can up-regulate the expression of MMP-2 ~ (8) ~ (11) ~ (11) ~ (16) ~ (19). The expression of TIMP-1 and PAI-1 was down-regulated and involved in the regulation of the proliferation of choriocarcinoma cells, which may be involved in the regulation of invasion and metastasis of choriocarcinoma.
【作者单位】：南方医科大学南方医院妇产科;
【基金】：国家自然科学基金(81170595)
【分类号】：R737.33
【正文快照】： 0引言F10基因(Gen Bank号:AB196290)是本研究从葡萄胎与正常早孕绒毛的差异c DNA文库中筛选出的一条功能未知的新基因[1]。研究表明F10基因在葡萄胎组织、侵蚀性葡萄胎、绒癌中均阳性表达且依次增强[2],提示F10与滋养细胞肿瘤的发生发展及侵袭和转移有关。细胞外基质和基底膜

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