华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

口腔鳞癌细胞过表达MHC-Ⅰ类链相关蛋白A对自然杀伤细胞与细胞毒性T淋巴细胞杀伤活性影响的实验研究

李超1 石芳琼1 杨丹1 王洁2 翦新春1 蒋灿华1   

  1. 1.中南大学湘雅医院口腔颌面外科; 2.中南大学湘雅医学院免疫学系, 长沙410078
  • 收稿日期:2012-02-25 修回日期:2012-02-25 出版日期:2012-02-01 发布日期:2012-02-01
  • 通讯作者: 蒋灿华,Tel:0731-84327475
  • 作者简介:李超(1985—),女,湖南人,硕士
  • 基金资助:

    国家自然科学基金资助项目(30772437);湖南省科技计划一般基金资助项目(06sk3026,06sk3044)

Natural killer and cytotoxic T lymphocyte-mediated cytotoxicity enhanced by genetic overexpression of MHC class Ⅰ chain-related protein A in oral squamous cell carcinoma:An experimental study in vivo

Li Chao1, Shi Fangqiong1, Yang Dan1, Wang Jie2, Jian Xinchun1, Jiang Canhua1   

  1. 1. Dept. of Oral and Maxillofacial Surgery, Xiangya Hospital, Central-south University, Changsha 410078, China; 2. Dept. of Immunology, Xiangya Medical School, Central-south University, Changsha 410078, China
  • Received:2012-02-25 Revised:2012-02-25 Online:2012-02-01 Published:2012-02-01
  • Contact: Jiang Canhua,Tel:0731-84327475
  • About author:Li Chao(1985—),女,湖南人,硕士

摘要:

目的探讨口腔鳞癌细胞过表达MHC-Ⅰ类链相关蛋白A(MICA)对自然杀伤细胞(NK)与细胞毒性T淋巴细胞(CTL)杀伤活性的影响。方法通过绘制细胞生长曲线、检测细胞周期、进行平板集落形成率及裸鼠皮下成瘤等方法对稳定转染并过表达MICA的口腔鳞癌细胞株进行生物学特性鉴定。采用乳酸脱氢酶释放法及流式细胞术分析口腔鳞癌细胞过表达MICA对NK与CTL细胞杀伤活性及自然杀伤细胞2族成员D(NKG2D)受体表达的影响。结果口腔鳞癌细胞转染MICA基因后主要生物学特性未发生改变,但能显著增强NK与CTL细胞杀伤活性并上调其表面NKG2D的表达,与未转染及转染空白载体的细胞比较,差异有统计学意义(P<0.05)。结论MICA可作为口腔鳞癌免疫基因治疗的潜在靶标,值得进一步研究。

关键词: 口腔, 鳞状细胞癌, MHC-Ⅰ类链相关蛋白A, 杀伤活性, 基因治疗

Abstract:

Objective To investigate the effect on natural killer(NK) and cytotoxic T lymphocyte(CTL)-mediated cytotoxicity by genetic overexpression of MHC class Ⅰ chain-related protein A(MICA) in oral squamous cell carcinoma (OSCC). Methods The OSCC cells by genetic overexpression of MICA were detected to identify the biological features including cell growth curve, cell cycle distribution, plate clone forming rate and tumorigenicity in nude mice. The expression of natural killer group 2, member D(NKG2D) receptor and the cytotoxicity to target tumor cells of NK92 and CTL cells, which co-cultured with the transfected OSCC cells or the non-transfected or blank vector-transfected controls, were measured by flow cytometry and lactate dehydrogenase(LDH) release assay. Results There was no difference in biological features before and after MICA gene transfection to OSCC cells. Flow cytometry and LDH release assay showed that MICA-overexpressed OSCC cells enhanced the cytotoxicity to target tumor cells and up-regulated the expression of NKG2D on NK92 and CTL(P<0.05). Conclusion MICA may be considered as a promising immunotherapy target of OSCC.

Key words: oral, squamous cell carcinoma, MHC class Ⅰ chain-related protein A, cytotoxicity, gene therapy