摘要
目的 :探讨Numb基因对结肠癌细胞迁移和侵袭的影响及其可能的作用机制。方法:应用蛋白质印迹法检测结肠上皮细胞HCEpi及结肠癌SW480、SW620和HCT-8细胞中Numb蛋白的表达。将含Numb过表达重组载体的慢病毒感染Numb表达水平较低的HCT-8细胞(HCT-8-Numb组),应用蛋白质印迹法检测空白对照组(HCT-8细胞未进行感染)、阴性对照组(HCT-8细胞感染含空载体的慢病毒)和HCT-8-Numb组HCT-8细胞中Numb蛋白的表达水平,并应用蛋白质印迹法、细胞划痕实验和Transwell法检测空白对照组、阴性对照组、HCT-8-Numb组和Ras相关C3肉毒素底物1(Ras-related C3 botulinum toxin substrate 1,Rac1)抑制剂NSC23766处理组HCT-8细胞中活化的Rac1蛋白的表达水平以及细胞迁移和侵袭能力。结果:结肠上皮细胞HCEpi中Numb蛋白的表达水平明显高于结肠癌SW480、SW620和HCT-8细胞(P值均<0.05),且HCT-8细胞中Numb蛋白的表达水平最低。HCT-8-Numb组细胞中Numb蛋白的表达水平明显高于空白对照组和阴性对照组(P值均<0.05)。HCT-8-Numb组细胞中活化的Rac1蛋白的表达水平明显低于空白对照组和阴性对照组(P值均<0.05),而与NSC23766处理组细胞中活化的Rac1蛋白的表达水平相比差异无统计学意义(P>0.05)。HCT-8-Numb组细胞的迁移和侵袭能力明显低于空白对照组和阴性对照组(P值均<0.05),而与NSC23766处理组细胞的迁移和侵袭能力相比差异无统计学意义(P>0.05)。结论 :过表达Numb蛋白可抑制结肠癌HCT-8细胞的迁移和侵袭,这一作用可能与抑制Rac1蛋白的活化有关。
Objective: To investigate the effects of Numb on invasion and migration of colorectal cancer cells, and to explore its possible mechanism.Methods: The expression levels of Numb in colonic epithelial cell line HCEpi and colorectal cancer cell lines SW480, SW620 and HCT-8 were detected by Western blotting. A recombinant lentiviral vector for Numb overexpression (LV-GV218-Numb-EGFP) was infected into HCT-8 cells with low expression of Numb (named as HCT-8-Numb group). The expression levels of Numb protein in the blank control (HCT-8 cells without any infection), negative control (HCT-8 cells infected with lentivirus containing an empty vector, LV-GV218-EGFP) and HCT-8-Numb groups were detected by Western blotting. The expression levels of the activated Ras-related C3 botulinum toxin substrate 1 (Rat1) and the abilities of migration and invasion of colonic epithelial cells and colorectal cancer cells in the blank control, negative control, HCT-8- Numb and NSC23766 (Racl inhibitor) treatment groups were detected by Western blotting, wound scratch assay and Transwell invasion assay, respectively.Results: The expression level of Numb protein in colonic epithelial cell line HCEpi was higher than those in colonic cancer cell lines SW480, SW620 and HCT-8 (P 〈 0.05). The Numb expression level in HCT-8 cells was lowest. The expression level of Numb protein in HCT- 8-Numb group was higher than those in the blank control and the negative control groups (both P 〈 0.05). The expression level of activated Racl protein in HCT-8-Numb group was significantly lower than those in the blank control and the negative control groups (both P 〈 0.05), but there was no significant difference between NSC23766 treatment group and HCT- 8-Numb group (P 〉 0.05). The abilities of migration and invasion in HCT-8-Numb group were decreased as compared with those in the blank control and the negative control groups (all P 〈 0.05), but there was no significant difference between NSC23766 treatment group and HCT-8-Numb group (P 〉 0.05).Conclusion: Numb overexpression can inhibit the invasion and migration of colonic cancer HCT-8 cells. This effect may be related to inhibiting the activation of Racl.
出处
《肿瘤》
CAS
CSCD
北大核心
2015年第8期842-848,共7页
Tumor