敲除KDM4B通过减少上皮间质转化抑制结肠癌转移与侵袭

KDM4B knockout inhibited metastasis and invasion of colorectal carcinoma through reducing epithelial - mesenchymal transition

目的 构建KDM4B敲除的人结肠癌HCT116和SW260细胞,研究KDM4B对结肠癌细胞迁移与侵袭的影响并探讨其机制。方法 利用免疫组化染色,检测临床组织样本KDM4B的表达量;利用CRISPR/Cas9技术,通过慢病毒侵染HCT116与SW260细胞,构建KDM4B敲除细胞系;利用细胞划痕实验及transwell小室实验评价KDM4B敲除细胞与野生型细胞迁移及侵袭能力的差异;利用Western blotting检测EMT标志蛋白E-cadherin、 N-cadherin和Vimentin的表达情况。结果 免疫组化表明结肠癌组织中KDM4B显著高表达;通过Western blotting成功验证了KDM4B敲除结肠癌细胞系;敲除KDM4B显著抑制HCT116及SW260细胞的迁移与侵袭;敲除KDM4B显著上调HCT116及SW260细胞中E-cadherin的表达,显著下调N-cadherin和Vimentin的表达。结论 KDM4B能够通过介导EMT促进结肠癌细胞HCT116和SW260的迁移与侵袭,将其敲除可以显著减弱这种作用。

Objective To construct KDM4B knockout human colon cancer HCT116 and SW260 cells, study the effect of KDM4B on migration and invasion of colon cancer cells, and explore its mechanism. Methods Immunohistochemical staining was used to detect the expression of KDM4B in clinical tissue samples;Using CRISPR/Cas9 technology, HCT116 and SW260 cells were infected by lentivirus to construct KDM4B knockout cell lines;The difference of migration and invasion between KDM4B knockout cells and wild type cells was evaluated by cell scratch test and transwell chamber test;Western blotting was used to detect the expression of E-cadherin, N-cadherin and Vimentin. Results Immunohistochemistry showed that KDM4B was highly expressed in colon cancer tissues;KDM4B knockout colon cancer cell line was successfully verified by Western blotting;Knockout of KDM4B significantly inhibited the migration and invasion of HCT116 and SW260 cells;Knockout of KDM4B significantly increased the expression of E-cadherin in HCT116 and SW260 cells, and significantly decreased the expression of N-cadherin and Vimentin. Conclusion KDM4B can promote the migration and invasion of colon cancer cells HCT116 and SW260 by mediating EMT, and its knockout can significantly reduce this effect.