敲低错配修复基因MSH6抑制结直肠癌细胞上皮间充质转化及其增殖

目的:探究错配修复基因MSH6对结直肠癌细胞的增殖、迁移和侵袭的影响及可能发生的机制。方法:根据美国国家生物信息技术中心(NCBI)中MSH6的基因序列构建靶向敲低MSH6的序列shMSH6-1、shMSH6-2和shMSH6-3，采用细胞转染技术敲低结直肠癌细胞中MSH6的表达，通过实时荧光定量 PCR 检测敲低效率并进行慢病毒包装，应用Western blot在细胞系中筛选MSH6高表达细胞系进行后续实验，应用CCK8检测细胞增殖能力，克隆形成实验检测细胞集落形成能力，伤口愈合和Transwell法检测细胞迁移和侵袭能力，通过Western blot方法检测上皮间充质相关蛋白的表达变化。结果:MSH6在结直肠癌细胞系中表达上调，其中RKO、SW620、LOVO细胞系上调明显，敲低MSH6明显限制了结直肠肿瘤细胞的增殖、迁移和侵袭，同时导致E-cadherin 蛋白水平增加，N-cadherin和Vimentin蛋白表达下降。结论:MSH6在结直肠癌中表达上调，敲低MSH6可抑制结直肠癌细胞的增殖、迁移和侵袭，诱导细胞发生凋亡并能够逆转EMT的发生。

Knockdown of mismatch repair gene MSH6 inhibits epithelial mesenchymal transition and its proliferation in colorectal cancer cells

Objective:To investigate the effect of mismatch repair gene MSH6 on proliferation,migration and invasion of colorectal cancer cells and the possible mechanism of occurrence.Methods:According to the MSH6 gene sequence of national center for biological information technology,the sequences shMSH6-1,shMSH6-2 and shMSH6-3 were constructed.Cell transfection was used to silence the expression of MSH6 in colorectal cancer cells,and the transfection effect was verified using PCR and Western Blot.CCK8,clone formation,wound healing,angiogenesis and transwell assay were applied to explore the effects of MSH6 on the biological functions of tumor cells.The expression of epithelial mesenchymal-associated proteins was verified by Western blot method.Results:MSH6 expression was up-regulated in colorectal cancer cell lines,especially in RKO,SW620 and LOVO cell lines.Knockdown of MSH6 significantly restricted the proliferation,migration and invasion of colorectal tumor cells,and led to the increase of E-cadherin protein level and the decrease of N-cadherin and Vimentin protein expression.Conclusion:The expression of MSH6 is up-regulated in colorectal cancer.Knockdown of MSH6 inhibited the proliferation,migration and invasion of colorectal cancer cells,induced apoptosis and was able to reverse the development of EMT.