摘要
目的:观察表皮生长因子受体单克隆抗体C225对人肺鳞癌细胞系(H-520)生长、凋亡和周期分布的影响。方法:流式细胞检测H-520细胞EGFR表达比例,将鼠抗人EGFR一抗和FITC标记的羊抗鼠二抗加入细胞悬液中,孵育、漂洗重悬细胞后流式检测。MTT法测定C225抑制H-520细胞生长最佳浓度和最佳时间,将不同浓度C225加入指数生长的细胞培养液中,继续培养一定时间(48h),检测细胞生长抑制率。流式细胞仪检测细胞凋亡以及细胞周期。结果:H-520细胞中EGFR表达比例高达82.25%。C225抑制H-520细胞生长的最佳浓度是40nmol/L,最佳作用时间是72h。细胞凋亡实验结果显示,H-520细胞自然凋亡率为5.56%±0.62%,C225可使其凋亡百分率上升到13.75%±0.83%(P<0.05)。细胞周期分布显示40nmol/L C225可使细胞阻滞于G0+G1期,S期细胞比例下降。结论:C225对H-520细胞生长抑制作用,可能与细胞G0+G1期阻滞后凋亡有关。
Objective：To investigate the effects of C225 （cetuximab）,a chimeric human-mouse anti-epithelial growth factor receptor monoclonal antibody,on the growth,apoptosis and cell cycle of human non-small cell lung cancer cell line H-520 and explore whether C225 treatment is appropriate for non-small cell lung cancer patients in clinics.Methods：With flow cytometric analysis,we first detected the expression level of EGFR on H-520 cells.To determine the appropriate concentration and time for C225 treatment,we treated H520 cells with C225 for 24,48 or 72 hours,respectively. In addition,H520 cells were treated with 40nmol/L C225 for 72 hours and thus harvested for flow cytometric analysis of the apoptosis percentage and cell cycle.Results： We found 82.25% of the H520 cells express EGFR.For the growth inhibition assay,40nmol/L and 72 hours were suitable for C225 treatment and used for the following apoptosis and cell cycle analysis.Moreover,C225 treatment significantly increased the apoptosis percentage of apoptotic H-520 cells （13.75%±0.83%） compared with the control group （5.56%±0.62%,P〈0.05）.The cell cycle analysis showed that C225 treatment led to cell cycle arrest in G0＋G1 phase,consequently,percentage of cells in S phase was decreased.Conclusion：C225 treatment significantly inhibits H520 cell growth which may be related with cell cycle arrest in G0＋G1 phase as well as the increased apoptosis,providing supportive evidence for clinical treatment of non-small cell lung cancer.
出处
《现代肿瘤医学》
CAS
2010年第9期1681-1683,共3页
Journal of Modern Oncology
