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目的探讨F10基因过表达及沉默对绒癌细胞系JEG-3侵袭相关蛋白酶表达的影响。方法采用细胞转染技术及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JEG-3。将SPF级裸鼠(4~5周龄)30只随机分为F10过表达组、F10沉默组和未处理组各10只,分别接种F10基因过表达的JEG-3细胞、F10基因沉默的JEG-3细胞和未处理的JEG-3细胞。于接种5周后处死裸鼠取皮下肿瘤,采用免疫组织化学法和Western blot法检测各组瘤体组织中基质金属蛋白酶(matrix metalloproteinase,MMP)、基质金属蛋白酶组织抑制物-1(tissue inhibitor of matrix metalloproteinase,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,并进行比较。结果免疫组织化学结果显示,F10过表达组成瘤组织中MMP-2、MMP-8、MMP-11、MMP-16、MMP-19蛋白表达水平(0.332±0.037、0.473±0.089、0.441±0.031、0.470±0.040、0.548±0.062)高于未处理组(0.209±0.021、0.314±0.108、0.207±0.025、0.400±0.057、0.466±0.038)和F10沉默组(0.049±0.013、0.168±0.034、0.149±0.024、0.240±0.024、0.175±0.029)(P<0.05),未处理组高于F10沉默组(P<0.05);F10过表达组TIMP-1(0.191±0.027)和PAI-1蛋白(0.130±0.020)表达水平低于未处理组(0.249±0.036、0.284±0.020)和F10沉默组(0.310±0.018、0.463±0.115)(P<0.05),未处理组低于F10沉默组(P<0.05)。wesrern blot检测结果显示,上述侵袭相关蛋白酶在3组间比较差异均有统计学意义(P<0.05)。结论 F10通过上调MMP-2、MMP-8、MMP-11、MMP-16、MMP-19表达,下调TIMP-1、PAI-1表达,促进绒癌细胞增殖、侵袭和转移。
Objective To investigate the effect of overexpression of F10 gene and silencing on the expression of protease related to invasion of choriocarcinoma cell line JEG-3. Methods Cell transfection technology and RNA interference technology were used to establish and screen the choriocarcinoma cell line JEG-3 with overexpression and silencing of F10 gene respectively. Thirty SPF nude mice (4 to 5 weeks old) were randomly divided into F10 overexpression group, F10 silencing group and untreated group, 10 mice in each group were inoculated with JEG-3 cells overexpressing F10 gene, JEG -3 cells and untreated JEG-3 cells. The tumors in nude mice were sacrificed 5 weeks after inoculation, and the expression of matrix metalloproteinase (MMP), tissue inhibitor of tissue inhibitor of metalloproteinase (TIMP-1) in each group were detected by immunohistochemistry and Western blot. TIMP-1) and plasminogen activator inhibitor-1 (PAI-1) were detected and compared. Results The results of immunohistochemistry showed that the expression of MMP-2, MMP-8, MMP-11, MMP-16 and MMP-19 in the overexpression group of F10 was significantly higher than that in the control group (0.332 ± 0.037, 0.473 ± 0.089, ± 0.040, 0.548 ± 0.062) were higher than those in the untreated group (0.209 ± 0.021,0.314 ± 0.108,0.207 ± 0.025,0.400 ± 0.057,0.466 ± 0.038) and F10 silencing group (0.049 ± 0.013,0.168 ± 0.034,0.149 ± 0.024, The levels of TIMP-1 (0.191 ± 0.027) and PAI-1 protein (0.130 ± 0.020) in F10 overexpression group were significantly higher than those in F10 silencing group (P <0.05) (P <0.05). The expression levels in the untreated group were lower than those in the F10 silencing group (0.249 ± 0.036,0.284 ± 0.020) and the F10 silencing group (0.310 ± 0.018,0.463 ± 0.115) (P <0.05). The results of wesrern blot showed that there were significant differences among the three groups in invasion-related proteases (P <0.05). Conclusion F10 can promote the proliferation, invasion and metastasis of choriocarcinoma cells by up-regulating the expression of MMP-2, MMP-8, MMP-11, MMP-16 and MMP-19 and down-regulating the expressions of TIMP-1 and PAI-