p75^NTRICD激活NF-κB对肿瘤细胞失巢凋亡抵抗的影响  

Influence of p75^(NTR) ICD on anoikis resistance of tumor cells through activating NF-κB

作　　者：鲍欣[1] 石剑波[1] 谢芙蓉[1] 徐骎[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院·口腔医学院,口腔颌面-头颈肿瘤科,上海市口腔医学重点实验室,上海200011

出　　处：《中国口腔颌面外科杂志》2017年第4期305-309,共5页China Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81272977)

摘　　要：目的 :通过构建肿瘤细胞失巢培养模型,探讨p75NTR ICD对肿瘤细胞失巢凋亡抵抗能力的影响及其相关信号通路的激活情况。方法:采用免疫组织化学方法观察p75NTR ICD在口腔鳞状细胞癌组织中的表达,Western免疫印迹方法检测高转移和低转移潜能口腔鳞癌细胞系中p75NTR ICD的表达。通过poly-HEMA(聚甲基丙烯酸羟乙基酯)包被培养皿,建立肿瘤细胞失巢培养模型。通过质粒转染方法,在细胞中过表达p75NTR ICD。采用流式细胞术检测肿瘤细胞在失巢培养条件下的凋亡水平,通过激光共聚焦显微镜及Western免疫印迹实验检测肿瘤细胞失巢培养模型中相关信号通路的激活情况。采用SPSS.22软件包对数据进行统计学分析。结果:发生淋巴结转移的口腔鳞状细胞癌原发灶组织标本中,p75NTR ICD定位于胞质中;未发生淋巴结转移的口腔鳞状细胞癌原发灶组织标本中,p75NTR ICD定位于胞膜上。高转移潜能HN-12细胞系中p75NTR ICD高表达,低转移潜能HN-4细胞中p75NTR ICD低表达,外源性过表达p75NTR ICD可降低HN-4细胞的失巢凋亡水平。与HN-4细胞相比,HN-12失巢培养后形成更大而圆的小球。HN-12细胞失巢凋亡水平低于HN-4细胞,高转移潜能HN-12细胞失巢培养后激活NF-κB,低转移潜能HN-4细胞失巢培养,NF-κB未能被激活。结论:p75NTR ICD在肿瘤细胞中的定位与肿瘤是否发生淋巴结转移相关。p75NTR ICD高表达肿瘤细胞具有失巢凋亡抵抗能力,p75NTR ICD通过激活NF-κB信号通路促进肿瘤细胞失巢凋亡抵抗。PURPOSE： To investigate the influence of p75^NIR ICD on anoikis resistance ability of tumor cells based on generation of an anoikis cell model and discuss the activation of related signaling pathways. METHODS： Immunohistochemistry was used to detect p75^NIR ICD expression in lymph node metastatic and non-lymph node metastatic, primary specimens of oral squamous cell carcinoma （OSCC）. Western blot was used to detect the expression of p75^NIR ICD in a pair of low- and high- metastatic potential OSCC cell lines. Poly-HEMA was used to pre-coat the cell culture dish in order to generate anoikis cell model. Over-expression of p75^NIR ICD was conducted in tumor cells with plasmid transfection. Flow cytometry was used to detect apoptosis level of tumor cells after suspension culture. Laser scanning confocal microscopy analysis and Western blot were performed to detect the activation of related signal pathways of tumor cells in anoikis cell model. SPSS 22.0 software package was used for statistical analysis. RESULTS： The positive staining of p75^NIR ICD in lymph node metastatic OSCC cells was located in the cytoplasm, while on the cytomembrane in non- lymph node metastatic OSCC. The expression of p75^NIR ICD was much higher in HN-12, which was more metastatic as well, compared with HN-4. Larger sphere was formed in HN-12 anoikis cell model than in HN-4, lower apoptosis level of HN-12 in anoikis cell model was observed than in HN-4. NF-κB signaling was activated in HN-12 anoikis cell model, while not in HN-α. CONCLUSIONS： Cellular localization of p75^NIR ICD in tumor cells was related to cervical lymph node metastasis. Over expression of p75^NIR ICD in tumor cells shows higher anoikis resistance; p75^NIR ICD promotes anoikis resistance through activating NF-κB signaling.

关 键 词：口腔鳞状细胞癌 失巢凋亡 P75^NTR ICD NF-ΚB 

分 类 号：R739.8[医药卫生—肿瘤]