本文關鍵詞： FOXQ1 上皮間質轉化(EMT) 大腸癌 轉移　出處：《重慶醫(yī)科大學》2015年碩士論文　論文類型：學位論文

【摘要】：目的：探討FOXQ1 (Forkhead Box Q1,FOXQ1)基因對大腸癌細胞的上皮-間質轉化(Epithelial-Mesenchymal transition, EMT)的作用及其可能的機制。方法：構建針對人類FOXQ1基因的shRNA慢病毒沉默載體,轉染大腸癌細胞株SW480細胞,并篩選出沉默效果最好的干擾序列。后續(xù)實驗設干擾組(感染FOXQ1-shl序列)、陰性組(感染NC-shRNA序列)、對照組。顯微鏡觀察三組細胞間形態(tài)學差異；Transwell小室、細胞黏附試驗觀察三組細胞轉移能力及黏附能力的差異；qRT-PCR、W estern blot技術檢測E-cadherin、N-cadherin、Vimentin、MMP2基因和蛋白水平的差異。結果：FOXQ1-sh1慢病毒載體成功沉默了SW480細胞的FOXQ1的表達。干擾組與陰性組及對照組相比,細胞頂底極性及細胞間緊密連接增加；侵襲、遷移的細胞數目顯著減少(P0.05)；同種黏附能力增加,異種黏附能力降低(P0.05)；qRT-PCR、Western blot顯示E-cadhe rin表達顯著增高而N-cadherin、Vimentin、MMP2表達顯著降低(P0.05)。結論：沉默FOXQ1基因能夠促進大腸癌SW480細胞EMT的逆轉,降低大腸癌SW480細胞的轉移能力,其機制可能與E-cadherin的上調和N-cadherin、Vimentin、MMP2的下調有關。
[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.

【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.34

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本文編號：1538798