背景:上皮间质转化(EMT)是恶性肿瘤细胞获得迁移和侵袭能力的重要生物学过程。本研讨摸索了KLF5在宫颈癌细胞株EMT进程中的感化。
目的:Krüpple样因子5(KLF5)是一种基本的转录因子,在细胞周期停滞和抑制凋亡中起关键作用。然而,KLF5介导宫颈癌细胞系生物学功能的分子机制尚未阐明。这里,我们重点研究了ELF5在宫颈癌细胞系体外模型中调节EMT过程的潜在功能。
方法:采用Westernblot和实时定量PCR检测HeLa细胞中EMT相关基因的表达。MTT测定,细胞划痕和Transwell试验用于评估HeLa细胞的增殖和侵袭能力。使用生物信息学工具JASPAR,我们在SNAI1基因启动子中鉴定出一个得分高的KLF5样结合序列.荧光素酶报告基因测定用于检测不同SNAI1启动子截短物的转录活性。
结果:在HeLa细胞中过表达KLF5基因后,KLF5不仅显著抑制HeLa细胞的侵袭和迁移,而且还增加了E-cadherin的表达,并降低了N-cadherin和MMP9的表达。此外,E-cadherin上游调节因子的mRNA表达,例如SNAI1,SLUG,ZEB1/2和TWIST1也降低。此外,KLF5通过结合其启动子区抑制SNAI1基因的表达,SNAI1基因过表达后促进Hela细胞的EMT。
结论:这些结果表明,KLF5可以通过降低SNAI1基因的表达来下调HeLa细胞的EMT过程,从而抑制HeLa宫颈癌细胞的迁移和侵袭。
UNASSIGNED: Epithelial-mesenchymal transition (EMT) is an important biological process by which malignant tumor cells to acquire migration and invasion abilities. This study explored the role of KLF5 in the EMT process of in cervical cancer cell lines.
UNASSIGNED: Krüpple-like factor 5 (KLF5) is a basic transcriptional factor that plays a key role in cell-cycle arrest and inhibition of apoptosis. However, the molecular mechanism by which KLF5 mediates the biological functions of cervical cancer cell lines has not been elucidated. Here, we focus on the potential function of ELF5 in regulating the EMT process in in vitro model of cervical cancer cell lines.
UNASSIGNED: Western-blot and real-time quantitative PCR were used to detect the expression of EMT-related genes in HeLa cells. MTT assays, cell scratch and Transwell assays were used to assess HeLa cells proliferation and invasion capability. Using the bioinformatics tool JASPAR, we identified a high-scoring KLF5-like binding sequence in the SNAI1 gene promoter. Luciferase reporter assays was used to detect transcriptional activity for different SNAI1 promoter truncates.
UNASSIGNED: After overexpressing the KLF5 gene in HeLa cells, KLF5 not only significantly inhibited the invasion and migration of HeLa cells, but also increased the expression of E-cadherin and decreased the expression of N-cadherin and MMP9. In addition, the mRNA expression of upstream regulators of E-cadherin, such as SNAI1, SLUG, ZEB1/2 and TWIST1 was also decreased. Furthermore, KLF5 inhibiting the expression of the SNAI1 gene via binding its promoter region, and the EMT of Hela cells was promoted after overexpression of the SNAI1 gene.
UNASSIGNED: These results indicate that KLF5 can downregulate the EMT process of HeLa cells by decreasing the expression of the SNAI1 gene, thereby inhibiting the migration and invasion of HeLa cervical cancer cells.