mAb-EGFR功能化修饰的纳米金棒靶向光热作用对下咽癌FADU细胞株和293T细胞株的温度影响

An Analysis of the Diversification of Cell Temperature of Photothermal Effects of Gold Nanorods Coated with mAb-EGFR on FADU Human Pharyngeal Squamous Cell Lines and Non-tumor Cells 293 T Cell Line

目的：探讨上皮生长因子受体单克隆抗体（epidermal grow th factor receptor monoclonal antibody ， mAb－EGFR）功能化修饰的纳米金棒靶向光热作用对下咽癌FADU细胞株和非肿瘤细胞的温度影响。方法在CTAB体系下合成实验所需纳米金棒并完成上皮生长因子受体（epidermal growth factor receptor ，EGFR）单克隆抗体功能化修饰，用紫外分光光度计及电镜表征；以体外培养的下咽癌FADU 细胞和非肿瘤细胞293T 细胞系为实验材料，用western blot免疫印迹法定性比较两株细胞EGFR的表达及明确细胞内吞纳米金棒；Annexin－5／PI双染法荧光显微镜观察FADU细胞和293T 细胞凋亡情况；设定不同剂量纳米金浓度（15％、25％、35％），应用808 nm波长近红外激光照射6分钟，每隔一分钟检测细胞培养介质温度，并用X T T 法检测细胞存活率。结果纳米金棒浓度在15％～25％、近红外激光照射6分钟时，细胞温度为41～43℃，并且趋于稳定，对下咽癌细胞有明显的杀伤作用（P＜0．05），而对293T 细胞没有明显的杀伤作用，而纳米金棒浓度在35％时对下咽癌 FADU细胞和293T细胞都具有明显的杀伤作用（P＜0．05）。结论浓度为15％～25％ mAb－EGFR功能化修饰的纳米金棒在近红外激光照射6分钟时细胞温度升至41～43 ℃，对下咽癌细胞具有明显的杀伤作用，而对非肿瘤细胞无明显损伤。

Objective To study the cell temperature on photothermal effects of gold nanorods coated with mAb-EGFR targeted on FADU human pharyngeal squamous cell lines and non- tumor cells（293T cell line） .Meth‐ods CTAB gold nanorods were synthesized to complete the functional modifications of EGFR monoclonal antibody , with a UV spectrophotometer and electron microscopy characterization .The cultured FADU hypopharyngeal cells and non-tumor cells 293 T cell lines were expressed by western blot EGFR statutory comparison of two cell lines , cells to swallow the gold nanorods were observed .Cell apoptosis was observed by Annexin-V -FLUOS and pro‐pidium iodide .Then we set different doses of gold nanoparticles concentration （15% ,25 % ,and 35% ） .After 6 minutes＇near-infrared laser irradiation （λ=808 nm） ,the temperature was recorded once every minute ,and cell viability was detected by XTT assay .Results After six minutes＇near -infrared laser irradiation ,the cell tempera‐ture was 41 ℃ ～43 ℃ ,with 15% ～25 % of gold nanorods .The cell killing effects on hypopharyngeal were signifi‐cant .The 293T cells did not show any damaging effects （P〈0 .05） ,whereas the concentration of gold nanoparticles in 35% of hypopharyngeal FADU cells and 293T cells had significant cytotoxicity （P〈0 .05） .Conclusion There were significant cell killing effects with 15% ～25% gold nanorods coated with mAb-EGFR in the near -infrared laser irradiation six minutes （Cell temperatures of both cell lines were 41 ℃～43 ℃）on hypopharyngeal FADU cell but not on non-tumor cells .