摘要
目的:检测Bmi1(B lymphoma Mo-MLV insertion region 1)在舌鳞癌细胞与正常舌黏膜中的表达差异,探讨Bmi1基因沉默后对口腔舌鳞癌细胞生物学功能的影响。方法:实时定量逆转录聚合酶链反应(q RT-PCR)和蛋白免疫印迹(Western blot)检测Bmi1蛋白和m RNA在舌鳞癌细胞系和正常舌黏膜组织中的表达情况;采用免疫荧光染色检测Bmi1在舌鳞癌细胞中的分布情况;通过短发夹sh RNA有效抑制Bmi1表达后,MTT实验检测其对舌鳞癌细胞增殖能力的影响;划痕实验检测其对舌鳞癌细胞迁移能力的影响;Transwell实验检测其对舌鳞癌细胞侵袭的影响;克隆形成实验检测其对舌鳞癌细胞克隆形成率的影响;构建移植瘤实验观察其对移植瘤生长的影响。结果:Bmi1在舌鳞癌细胞中高表达,而在正常舌黏膜中低表达(P<0.05);Bmi1基因沉默后显著抑制舌鳞癌细胞的增殖、侵袭、迁移能力以及抑制体内移植瘤的生长(P<0.05),这与Bmi1对下游靶基因P16、P14和E-cadherin的调控有关。结论 :Bmi1与舌鳞癌多种恶性生物学功能的调控密切相关,可能是舌鳞癌潜在的治疗靶点。
Objective：To investigate both m RNA and protein levels of B lymphoma Mo-MLV insertion region 1（Bmi1） in a panel of tongue squamous cell carcinoma（TSCC） cell lines as compared with normal tongue mucosa, and then to study biological roles of Bmi1 in tongue tumorigenesis by loss-of-function assays using small interference RNA. Methods： RNA and protein expressions of Bmi1 in TSCC cell lines and normal tongue mucosa were detected by q RT-PCR and Western blot. Cellular immunofluorescence was performed to further characterize the subcellular distribution of Bmi1 in tongue cancer cell lines. Cell migration, invasion,proliferation and colony formation were assessed by wound-healing, Transwell, MTT and colony-forming experiments. To further reinforce the notion that Bmi1 is critical for tongue cancer growth in vivo, genetic approach was further utilized to inhibit Bmi1 in a tongue cancer xenograft model. Results： Bmi1 m RNA and protein levels in TSCC cell lines were significantly higher than that in normal tongue mucosa as assessed by real-time RT-PCR and Western blot assays（P 〈 0.05）. Short-hairpin RNA-mediated Bmi1 knockdown inhibited cell proliferation, migration and invasion, reduced colony formation, presumably by modulation of p16, p14 and E-cadherin. Short-hairpin RNA-mediated Bmi1 knockdown significantly impaired tumor growth in a tongue cancer xenograft model. Conclusion：Bmi1 may serve as a key driver with multiple biological functions during tongue cancer progression and a novel therapeutic target against tongue cancers.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2016年第1期39-45,50,共8页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省自然科学基金(BK20151561)
