摘要
目的探讨microRNA-375(miR-375)对结直肠癌(CRC)细胞侵袭迁移的影响及该影响与Notch2通路蛋白的关系。方法将表达miR-375的慢病毒转染入人CRC细胞HCT-116细胞中作为实验组,将空载慢病毒转染入HCT-116细胞中作为阴性对照组(NC组),未转染HCT-116细胞作为空白组,采用实时荧光定量PCR(qRT-PCR)方法检验转染效率,Transwell小室实验检测细胞侵袭和迁移能力,Western Blot方法检测Notch2通路蛋白的相对表达。再将Notch2小干扰RNA(siRNA)转染入HCT-116细胞中作为Notch2-siRNA组,将阴性对照转染入HCT-116细胞中作为Notch2阴性对照组,未转染HCT-116细胞作为空白组,采用Western Blot方法检测3组细胞Notch2蛋白表达量,采用Transwell小室实验进一步验证Notch2表达的改变对CRC细胞侵袭迁移的影响。结果 CRC细胞中miR-375表达明显低于人正常结直肠黏膜细胞(t=33.00,P<0.05)。使HCT-116细胞中的miR-375过表达后,与NC组和空白组相比,细胞的侵袭、迁移能力均下降(F=223.40～894.16,P<0.05),且Notch2通路蛋白相对表达量下降(F=100.63,P<0.05)。将Notch2下调后,与Notch2阴性对照组和空白组相比,Notch2-siRNA组细胞的侵袭迁移能力下降(F=144.31～197.54,P<0.05)。结论上调miR-375表达可以抑制CRC细胞的侵袭迁移,这一作用可以通过调控Notch2来实现。
Objective To investigate the effect of microRNA-375 (miR-375) on the invasion and migration of colorectal cancer (CRC) and its association with Notch2 pathway protein. Methods Human CRC HCT-116 cells transfected with the lentivirus with miR-375 expression were included as experimental group, HCT-116 cells transfected with the empty lentivirus were included as negative control group (NC group), and untransfected HCT-116 cells were included as blank group. Quantitative real-time PCR was used to measure transfection efficiency, Transwell chamber was used to evaluate cell invasion and migration, and Western Blot was used to measure the relative expression of Notch2 pathway protein. HCT-116 cells transfected with Notch2 small interfering RNA (siRNA) were included as Notch2-siRNA group, HCT-116 cells transfected with negative control were included as Notch2 negative control group, and untransfected HCT-116 cells were included as blank group. Western Blot was used to measure the protein expression of Notch2, and Transwell chamber was used to verify the effect of the change in Notch2 expression on the invasion and migration of CRC. Results CRC cells had significantly lower expression of miR-375 than normal human colorectal mucosal cells ( t =33.00, P <0.05). Compared with the NC group and the blank group, the HCT-116 cells with miR-375 overexpression had significant reductions in the invasion and migration of cells ( F =223.40-894.16, P <0.05) and the relative expression of Notch2 pathway protein ( F =100.63, P <0.05). Compared with the Notch2 negative control group and the blank group, the Notch2-siRNA group with downregulated Notch2 had significant reductions in the invasion and migration of cells ( F =144.31-197.54, P <0.05). Conclusion Upregulation of miR-375 expression can inhibit the invasion and migration of CRC cells, which can be achieved by the regulation of Notch2.
作者
黄巧
孟海宁
任静
罗路
齐宏
沈若武
HUANG Qiao;MENG Haining;REN Jing;LUO Lu;QI Hong;SHEN Ruowu(Medical Aspects of Specific Environments, Qingdao University, Qingdao 266021, China)
出处
《青岛大学学报（医学版）》
CAS
2019年第2期211-215,共5页
Journal of Qingdao University(Medical Sciences)
基金
山东省高等学校科技计划项目(L16LK04)
