MiR-200c调节人结肠癌SW480细胞迁移与侵袭的分子机制研究  

Molecular mechanisms of miR-200c regulating invasion and migration of human colon cancer SW480 cells

作　　者：何平[1] 宋辉[1] 梁杰雄[1] 邵天松[1] 张钊[1] 李洋[1] He Ping;Song Hui;Liang Jiexiong;Shao Tiansong;Zhang Zhao;Li Yang(Department of General Surgery, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China)

机构地区：[1]首都医科大学附属北京安贞医院普外科

出　　处：《实用肿瘤杂志》2019年第3期210-214,共5页Journal of Practical Oncology

基　　金：国家自然科学基金(30972886)

摘　　要：目的探究miR-200c对人结肠癌SW480细胞侵袭和迁移的影响及其相关机制。方法将SW480细胞分为类似物转染组(转染miR-200c类似物)、抑制物转染组(转染miR-200c核酸抑制剂)、类似物阴性对照组(转染miR-200c类似物的无义序列)、抑制物阴性对照组[加入miR-200c核酸抑制剂的载体(不含miR-200c核酸抑制剂)]以及空白组(不进行任何处理)。实时定量聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)检测12 h和24 h转染效率。细胞划痕实验和transwell侵袭实验检测细胞迁移和侵袭。Western blot法检测转染后β-catenin和E-cadherin的表达。结果细胞划痕实验显示,类似物转染组12 h和24 h后伤痕宽度均高于类似物阴性对照组(均P<0.05),抑制物转染组均低于抑制物阴性对照组(均P<0.05)。Transwell实验显示,与类似物阴性对照组和抑制物阴性对照组比较,类似物转染组24 h和48 h通过8μm孔径的细胞数减少,抑制物转染组增多(均P<0.05)。类似物转染组β-catenin和E-cadherin的表达均升高,抑制物转染组各蛋白表达均下降(均P<0.05)。结论 miR-200c可能通过Wnt/β-catenin信号通路抑制SW480细胞的侵袭和迁移。Objective To explore the effect of miR-200 c on the invasion and migration of human colon cancer SW480 cells. Methods SW480 cells were divided into the analogue transfection group(SW480 cells transfected with miR-200 c mimics), inhibitor transfection group(SW480 cells transfected with miR-200 c nucleic acid inhibitor), analogue negative control group(SW480 cells transfected with miR-200 c mimics nonsense sequence), inhibitor negative control group(SW480 cells added with miR-200 c nucleic acid inhibitor carrier, without miR-200 c nucleic acid inhibitors), and blank group(SW480 cells without any treatment). Real-time polymerase chain reaction(RT-PCR) was used to detect the transfection efficiency at 12 h and 24 h. Scratch experiments and transwell assays were used to detect cell migration and invasion. Western blot was used to detect the expression of β-catenin and E-cadherin after transfection. Results Scratch experiments showed that the wound widths at 12 h and 24 h of the analogue transfection group were higher than those in the analogue negative control group(both P<0.05);and those of the inhibitor transfection group were lower than those of the inhibitor negative control group(both P<0.05). Transwell assays showed that, compared with the analogue negative control and inhibitor negative control groups, the number of cells passing through the 8 μm pore was significantly decreased at 24 h and 48 h in the analogue transfection group, and was significantly increased in the inhibitor transfection group(all P<0.05). The expressions of β-catenin and E-cadherin were increased in the analogue transfection group, and the expression of each protein in the inhibitor transfection group was decreased significantly(all P<0.05). Conclusion miR-200 c can inhibit the invasion and migration of SW480 cells through Wnt/β-catenin signaling pathway.

关 键 词：结肠肿瘤/病理学 微RNAs 细胞运动 肿瘤侵润 Wnt蛋白质类/代谢 β连环素/代谢 信号传导 肿瘤细胞 培养的 

分 类 号：R735.35[医药卫生—肿瘤] R730.23[医药卫生—临床医学]