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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si Grb7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。
Objective: To investigate the effect of growth factor receptor-bound 7 (Grb7) on the growth of oral squamous cell carcinoma and xenograft in nude mice. Methods: hNOK and Cal27 cells were cultured and the expression of Grb7 protein was detected by real-time PCR and Western blot. Grb7 was silenced by siRNA and transfected into Cal27 cells for 48 h. Cell proliferation was measured by MTT assay. Cell cycle and apoptosis rate were detected by flow cytometry. Western blotting was used to detect the protein expressions of Cyclin D1, Rb, E2F1, Caspase3, Bax and Bcl- The effect of silencing Grb7 on ERK1 / 2 and FOXM1 protein expression was also examined. Cal27 cells were transfected into si Grb7 and inoculated into nude mice to establish oral squamous cell carcinoma xenograft model and to measure the size and weight of the xenografts. Data were analyzed statistically using SPSS 11.0 software package. RESULTS: Grb7 was overexpressed in Cal27 cells. Silencing of Grb7 significantly inhibited cell proliferation, blocked G1 / S phase transition and promoted apoptosis. Silencing Grb7 downregulated the expression of Cyclin D1, Rb and Bcl-2, up-regulated the expression of E2F1, c-caspase3 and Bax. Knockdown of Grb7 significantly inhibited the expression of p-ERK1 / 2 and FOXM1. In addition, the size and weight loss of silencing Grb7 xenografts in nude mice was reduced. Conclusion: Grb7 inhibits the proliferation of Cal27 cells via ERK / FOXM1 pathway, promotes apoptosis and reduces the growth of xenografts.