中国口腔颌面外科杂志 ›› 2017, Vol. 15 ›› Issue (4): 305-309.

• 论著 • 上一篇    下一篇

p75NTR ICD激活NF-κB对肿瘤细胞失巢凋亡抵抗的影响

鲍欣, 石剑波, 谢芙蓉, 徐骎   

  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
  • 出版日期:2017-08-10 发布日期:2017-08-14
  • 通讯作者: 徐骎,E-mail:xuqin_2004@hotmail.com
  • 作者简介:鲍欣(1991-),女,硕士,E-mail:baoxin396@sina.com

Influence of p75NTR ICD on anoikis resistance of tumor cells through activating NF-κB

BAO Xin, SHI Jian-bo, XIE Fu-rong, XU Qin.   

  1. Department of Oromaxillofacial Head Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Stomatology. Shanghai 200011, China
  • Online:2017-08-10 Published:2017-08-14

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。

关键词: 口腔鳞状细胞癌, 失巢凋亡, p75NTR ICD, NF-κ, B

Abstract: PURPOSE: To investigate the influence of p75, ICD on anoikis resistance ability of tumor cells based on generation of an anoikis cell model, discuss the activation of related signaling pathways. METHODS: Immunohistochemistry was used to detect p75, ICD expression in lymph node metastatic, non-lymph node metastatic, primary specimens of oral squamous cell carcinoma. Western blot was used to detect the expression of p75, ICD in a pair of low-, 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, 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, Western blot were performed to detect the activation of related signal pathways of tumor cells in anoikis cell model. SPSS22.0software package was used for statistical analysis. RESULTS: The positive staining of p75NTR 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, ICD was much higher in HN-12, which was more metastatic as well, compared with HN-4. Larger sphere was formed in HN-12anoikis cell model than in HN-4, lower apoptosis level of HN-12in anoikis cell model was observed than in HN-4. NF-κB signaling was activated in HN-12anoikis cell model, while not in HN-4. CONCLUSIONS: Cellular localization of p75, ICD in tumor cells was related to cervical lymph node metastasis. Over expression of p75NTR ICD in tumor cells shows higher anoikis resistance; p75, ICD promotes anoikis resistance through activating NF-κB signaling

Key words: Oral squamous cell carcinoma, Anoikis, p75NTR ICD, NF-κB

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