FOXQ1基因在大肠癌SW480细胞的EMT中的作用

发布时间：2018-02-26 16:32

本文关键词： FOXQ1 上皮间质转化(EMT) 大肠癌转移　出处：《重庆医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：探讨FOXQ1 (Forkhead Box Q1,FOXQ1)基因对大肠癌细胞的上皮-间质转化(Epithelial-Mesenchymal transition, EMT)的作用及其可能的机制。方法：构建针对人类FOXQ1基因的shRNA慢病毒沉默载体,转染大肠癌细胞株SW480细胞,并筛选出沉默效果最好的干扰序列。后续实验设干扰组(感染FOXQ1-shl序列)、阴性组(感染NC-shRNA序列)、对照组。显微镜观察三组细胞间形态学差异；Transwell小室、细胞黏附试验观察三组细胞转移能力及黏附能力的差异；qRT-PCR、W estern blot技术检测E-cadherin、N-cadherin、Vimentin、MMP2基因和蛋白水平的差异。结果：FOXQ1-sh1慢病毒载体成功沉默了SW480细胞的FOXQ1的表达。干扰组与阴性组及对照组相比,细胞顶底极性及细胞间紧密连接增加；侵袭、迁移的细胞数目显著减少(P0.05)；同种黏附能力增加,异种黏附能力降低(P0.05)；qRT-PCR、Western blot显示E-cadhe rin表达显著增高而N-cadherin、Vimentin、MMP2表达显著降低(P0.05)。结论：沉默FOXQ1基因能够促进大肠癌SW480细胞EMT的逆转,降低大肠癌SW480细胞的转移能力,其机制可能与E-cadherin的上调和N-cadherin、Vimentin、MMP2的下调有关。
[Abstract]:Objective: To investigate the FOXQ1 (Forkhead Box Q1, FOXQ1) - gene on epithelial mesenchymal transition in human colon cancer cells (Epithelial-Mesenchymal, transition, EMT) function and its possible mechanism. Methods: to construct the shRNA lentiviral vector targeting human FOXQ1 gene silencing, transfected into colon cancer cell line SW480, and filter out the interference sequence silence effect the best. The subsequent experiment set interference group (infected with FOXQ1-shl sequence), negative group (control group, NC-shRNA infection sequence). To observe the morphological differences between the three groups of cells microscope; Transwell cell, cell adhesion test differences were observed in the three groups cell metastasis and adhesion; qRT-PCR, E-cadherin W estern blot detection, N-cadherin technology. Vimentin, MMP2 gene and protein level. Results: FOXQ1-sh1 lentiviral vector was successfully silenced the expression of SW480 FOXQ1 cells. The interference group and the negative group and control group Compared to the top and bottom cell tight junctions between cells increased; the number of invasion, cell migration was significantly reduced (P0.05); with the increasing of adhesion ability, reduce the heterogeneous adhesion ability (P0.05); qRT-PCR Western, blot E-cadhe showed that Rin expression was significantly increased while N-cadherin, Vimentin, MMP2 expression decreased significantly (P0.05). Conclusion: the silence FOXQ1 gene can promote human colorectal cancer cell line SW480 EMT reversal, reduce the metastatic ability of human colorectal cancer cell line SW480 and its possible mechanism of Vimentin and the upregulation of E-cadherin and N-cadherin, and down-regulation of MMP2.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.34

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