Bmi1对舌鳞癌侧群细胞迁移、侵袭和增殖能力的调控作用及机制探讨

Bmi1 mediated migration, invasion and proliferation of stem cells of tongue squamous cell carcinoma

目的 :探讨Bmi1如何调控舌鳞癌侧群细胞的迁移、侵袭和增殖能力。方法 :首先采用Hoechst33342法,利用流式细胞仪分选舌鳞癌细胞株UM1(高转移株)中的侧群细胞(side population cell,SP)和非侧群细胞(non-side population cell,Non-SP)。Transwell实验检测沉默Bmi1后SP细胞的迁移、侵袭能力。球囊形成和平板克隆实验检测沉默Bmi1后SP细胞的球囊和克隆形成率;CCK8实验检测沉默Bmi1后SP细胞的增殖能力。Western免疫印迹检测沉默Bmi1后SP细胞中侵袭、转移相关基因(SOD2、Slug)及干细胞标志物(ABCG2、Nanog)的表达水平。采用SPSS17.0软件包对数据进行统计学处理。结果:转染Bmi1 si RNA使SP细胞中Bmi1表达水平下调后,SP细胞的迁移和侵袭能力显著下降;球囊形成率和克隆形成率也显著低于对照组;增殖能力受到抑制;SOD2、Slug和干细胞标志物ABCG2和Nanog的表达水平显著下降。结论 :沉默Bmi1可调控舌鳞癌侧群细胞的迁移、侵袭和增殖。

PURPOSE： To investigate the mechanism of Bmi1 mediating migration, invasion and proliferation of stem cells of tongue squamous cell carcinoma（TSCC）. METHODS： SP（side population cell, SP） and non-SP（non-side population cell） cells were sorted by FCM using Hoechst33342 method. Transwell assay was performed to detect migration and invasion of SP cells after Bmi1 knockdown. Sphere forming assay was conducted to detect sphere forming rate of SP cells after Bmi1 knockdown. Colony forming assay was used to detect colony forming rate of SP cells after Bmi1 knockdown.CCK8 assay was used to detect proliferation of SP cells after Bmi1 knockdown. Western blot assay was used to detect expression of invasion and metastasis related genes（SOD2 and Slug） and stem cell markers（ABCG2 and Nanog） in SP cells after Bmi1 knockdown. The data was analyzed using SPSS 17.0 software package. RESULTS： After knockdown of Bmi1 in SP cells, migration and invasion were significantly inhibited compared to control si RNA transfection. SP cells transfected with Bmi1 si RNA displayed significantly decreased sphere formation and colony formation compared to the cells transfected with control si RNA. The proliferation of SP cells was significantly inhibited after transfection with Bmi1 si RNA. The expression of SOD2, Slug and stem cell markers（ABCG2, Nanog） in SP cells were significantly decreased after transfection with Bmi1 si RNA. CONCLUSIONS： Bmi1 knockdown inhibited migration, invasion and proliferation of cancer stem cells in TSCC.