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目的:利用反义核酸技术探讨纤黏连蛋白(fibronectin, FN)诱导Hela细胞中基质金属蛋白酶(matrix metalloproteinases, MMPs)基因表达的机制。方法:无血清培养Hela细胞,以不同浓度及作用时间的FN诱导Hela细胞中MMPs基因表达, 并用反义寡聚脱氧核苷酸(oligodeoxynucleotides,ODN)封闭焦点黏着激酶(focal adhesion kinase,FAK), 用酶谱分析方法检测MMPs的活性。结果:FN浓度为0 μg/mL时,Hela细胞无MMP蛳2基因表达;FN浓度为5 μg/mL~10 μg/mL、作用时间为12 h,MMP蛳2活性最大;当在培养液反义ODN后,MMP蛳2活性明显降低。结论:MMP蛳2基因表达与FN的浓度以及作用时间有关;FN可通过其整合素受体的信号转导通路启动Hela细胞中MMP蛳2基因表达、降解ECM,从而促进肿瘤的浸润和转移。
OBJECTIVE: To explore the mechanism of matrix metalloproteinases (MMPs) gene expression induced by fibronectin (FN) in Hela cells by using antisense oligonucleotide technique. Methods: Hela cells were cultured in serum - free medium. The expression of MMPs gene was induced by FN at different concentrations and time, and focal adhesion kinase (FAK) was blocked by oligodeoxynucleotides (ODN) The activity of MMPs was detected by zymography. RESULTS: No mRNA level of MMP 蛳 2 was detected in Hela cells when FN concentration was 0 μg / mL. FN concentration was 5 μg / mL ~ 10 μg / mL for 12 h, and MMP 蛳 2 activity was the highest. After ODN, the activity of MMP 蛳 2 decreased significantly. Conclusion: The expression of MMP 蛳 2 gene is related to the concentration of FN and its action time. FN can promote the invasion and metastasis of HeLa cells through its signal transduction pathway of integrin receptor.