上海口腔医学 ›› 2016, Vol. 25 ›› Issue (6): 688-693.

• 论著 • 上一篇    下一篇

沉默Grb7对口腔鳞癌细胞Cal27生物学行为的影响

刘秉尧, 曹罡, 董震, 陈伟, 徐金科, 张森林, 翁志强   

  1. 南京军区南京总医院 口腔科,江苏 南京 210002
  • 收稿日期:2016-03-29 出版日期:2016-12-25 发布日期:2016-12-29
  • 通讯作者: 翁志强,E-mail:Wengzhiqiang9898@sina.com
  • 作者简介:刘秉尧(1982-),男,硕士,主治医师,E-mail: LBY820122@126.com

Knockdown of Grb7 inhibits growth of oral squamous cell carcinoma, cell proliferation and promoted apoptosis through ERK/FOXM1 pathway

LIU Bing-yao, CAO Gang, DONG Zhen, CHEN Wei, XU Jin-ke, ZHANG Sen-lin, WENG Zhi-qiang   

  1. Department of Stomatology, Nanjing General Hospital of Nanjing Military Command. Nanjing 210002, Jiangsu Province, China
  • Received:2016-03-29 Online:2016-12-25 Published:2016-12-29
  • Contact: 英文通讯作者
  • About author:英文作者简介
  • Supported by:
    英文基金

摘要: 目的研究生长因子受体结合蛋白7(growth factor receptor-bound 7,Grb7)对口腔鳞癌细胞生长及裸鼠移植瘤的作用。方法培养hNOK和Cal27细胞,实时定量PCR和Western印迹法检测Grb7的蛋白表达。采用siRNA沉默Grb7并转染至Cal27细胞48 h,MTT法检测细胞增殖,流失细胞仪检测细胞周期和凋亡率,Western印迹法检测Cyclin D1、Rb、E2F1、Caspase3、Bax和Bcl-2的蛋白表达;同时检测沉默Grb7对ERK1/2和FOXM1蛋白表达的影响。Cal27细胞经转染siGrb7后接种于裸鼠,建立口腔鳞癌移植瘤模型,测量移植瘤体积及重量。采用SPSS 11.0软件包对数据进行统计学分析。结果Grb7在Cal27细胞中高表达,沉默Grb7显著抑制细胞增殖、阻滞G1/S期转换、促进细胞凋亡。沉默Grb7下调Cyclin D1、Rb和Bcl-2表达,上调E2F1、c-caspase3和Bax表达。敲减Grb7显著抑制p-ERK1/2和FOXM1的表达。此外,沉默Grb7裸鼠移植瘤的体积及重量减少。结论Grb7通过ERK/FOXM1通路抑制Cal27细胞增殖、促进凋亡并减少移植瘤增长。

关键词: 口腔鳞癌, 生长因子受体结合蛋白7, 细胞增殖, 细胞凋亡, 移植瘤

Abstract: PURPOSE: To investigate the effect of growth factor receptor-bound 7 (Grb7) on oral squamous cell carcinoma growth and tumor xenografts. METHODS: Cal27 and hNOK cells were cultivated, real-time PCR and Western blotting were used to detect the expression of Grb7 in hNOK and Cal27. Cal27 was transfected with Grb7 siRNA for 48 h, cell proliferation was assayed using MTT. Flow cytometry was used to determine the cell cycle and apoptosis. Western blot was used to evaluate the expression of caspase3, Bax, bcl-2, Cyclin D1, Rb, E2F1, ERK and FOXM1. Grb7 siRNA and negative control were designed and injected subcutaneously into the mice, tumor volume and weight were measured. Statistical analysis was performed using SPSS 11.0 software package. RESULTS: Grb7 was highly expressed in Cal27 compared with hNOK. Depletion of Grb7 significantly inhibited cell proliferation, blocked G1/S phase transition, promoted cell apoptosis. Knockdown of Grb7 suppressed the expression of Cyclin D1 and Rb, upregulated E2F1 expression. Moreover, c-caspase 3 and Bax was also reduced after inhibition of Grb7. ERK/FOXM1 signaling pathway was also inhibited by Grb7. In addition, the volume and weight of tumor xenografts were reduced by siGrb7. CONCLUSIONS: Depletion of Grb7 inhibits cell proliferation, promotes apoptosis and reduces tumor xenografts through ERK/FOXM1 pathway.

Key words: Oral squamous cell carcinoma, Growth factor receptor-bound 7 (Grb7), Cell proliferation, Cell apoptosis, Tumor xenografts

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