α6整合素通过激活TGFβ/Smad2通路促进结直肠癌迁移与侵袭
| 作者: |
1,2,3祝子雯,
4詹娜,
1,2,3董卫国
1 武汉大学人民医院消化内科，武汉 430060 2 武汉大学人民医院消化系统疾病湖北省重点实验室，武汉 430060 3 武汉大学人民医院中心实验室，武汉 430060 4 武汉大学人民医院病理科，武汉 430060 |
| 通讯: |
董卫国
Email: dongweiguo@whu.edu.cn |
| DOI: | 10.3978/j.issn.2095-6959.2018.04.004 |
摘要
目的：探讨α6整合素(integrin α6，ITGA6)在结直肠癌(colorectal cancer，CRC)中的表达及其对迁移与侵袭的影响。方法：使用生物信息分析ITGA6在CRC中的表达，Western印迹法检测其在CRC组织及细胞中的表达；使用瞬时转染方法建立ITGA6过表达SW480及HCT116细胞及对照细胞株；Traswell迁移及侵袭实验检测ITGA6对CRC细胞迁移及侵袭能力的影响。于倒置显微镜下观察CRC细胞形态的改变；Western印迹法及免疫荧光检测ITGA6过表达细胞及对照细胞中上皮-钙黏素(E-cadherin)，神经-钙黏素(N-cadherin)及波形蛋白(vimentin)的表达；基因集合富集分析(gene set enrichment analysis，GSEA)与ITGA6相关的通路并通过Western印迹法验证。结果：ITGA6在CRC组织和细胞中的表达明显高于对照组织及肠上皮永生化细胞；过表达ITGA6后，SW480和HCT116细胞从圆形、短梭形变得更为细长，整个细胞形态向长梭形转变。Transwell结果显示过表达ITGA6可促进CRC细胞迁移及侵袭能力；Western印迹法及免疫荧光显示过表达ITGA6后，E-cadherin表达下调，N-cadherin及vimentin的表达上调；GSEA发现ITGA6的过表达可上调TGFβ通路表达，Western印迹法检测显示过表达ITGA6可激活TGFβ/Smad2通路。结论：ITGA6在CRC中高表达，并可通过激活TGFβ/Smad2通路促进CRC细胞上皮-间质转化和肿瘤转移。
关键词：
结直肠癌；α6整合素；EMT；TGFβ/Smad2通路
Effect of integrin α6 on migration and invasion ability of colorectal cancer via TGFβ/Smad2 pathway
CorrespondingAuthor: DONG Weiguo Email: dongweiguo@whu.edu.cn
DOI: 10.3978/j.issn.2095-6959.2018.04.004
Abstract
Objective: To investigate the expression of integrin α6 (ITGA6) in colorectal cancer and explore the effect of ITGA6 on migration and invasion ability. Methods: Bioinformatics was used to analysis the expression of ITGA6 in colorectal cancer, Western blot was chosen to detect ITGA6 expression in colorectal cancer specimens and cell lines, transient transfection of ITGA6 and control plasmid was selected to construct SW480, HCT116 ITGA6 overexpressed cells and control cells, Transwell assays were used to detect colorectal cancer (CRC) cell migration and invasion ability, inverted microscope was used to analysis the morphology of cultured cell, the effect of ITGA6 overexpression on epithelial-mesenchymal transition (EMT) markers (E-cadherin, N-cadherin, vimentin) was obtained by Western blot and immunofluorescence assays. Gene set enrichment analysis (GSEA) was selected to explore ITGA6 related pathway and Western blot was further used to verify. Results: ITGA6 expression level was significantly higher in CRC tissues and CRC cells compared with paired normal mucosa and normal mucosa cell, after ITGA6 overexpression, morphology of SW480 and HCT116 cells changed from spheroid to spindle; Transwell assays showed ITGA6 overexpression enhanced migration and invasion ability; Western blot and immunofluorescence results showed ITGA6 overexpression reduced epithelial marker E-cadherin and upregulated mesenchymal markers N-cadherin, vimentin, GSEA analysis showed ITGA6 was positively related with TGFβ pathway, Western blot results confirmed ITGA6 could activate TGFβ/Smad2 pathway. Conclusion: ITGA6 was overexpresses in colorectal cancer and modulates TGFβ/SMAD2 pathway to induce EMT and promote metastasis.
Keywords：
colorectal cancer; integrin α6; epithelial-mesenchymal transition; TGFβ/Smad2 pathway