【作者】 张敏；

【导师】 王雪峰；

【作者基本信息】 辽宁医学院 ， 耳鼻咽喉科学， 2014， 硕士

【摘要】 目的探讨树突状细胞（DCs）在体外诱导、激活的T淋巴细胞与西妥昔单抗对头颈部鳞状细胞癌（HNSCC）杀伤作用，进一步指导临床治疗，提高HNSCC患者的生存率以及生活质量。方法用细胞因子rhIL-4、rhGM-CSF与rhTNF-在体外诱导扩增人外周血来源的单个核来源的DCs细胞，将DCs诱导的T淋巴细胞与药物西妥昔单抗联合作用于HNSCC，HNSCC选用UV-SCC-10A细胞株，用MTT法检测两者对UV-SCC-10A杀伤率的影响，流式细胞仪检测其凋亡率的影响，并用Western blot法检测各组EGFR与pEGFR的表达。各组数据用SPSS17.0软件进行统计学分析。结果DCs在体外由诱导培养成功，倒置显微镜下观察DCs的形态典型。流式细胞术检测结果证实其表型CD80，CD86表达率在50％左右，符合报道；MTT结果显示，随着西妥昔单抗药物浓度逐渐增大，其对UV-SCC-10A的抑制作用逐渐增强，并且T淋巴细胞能明显增强西妥昔单抗对UV-SCC-10A的杀伤率，其差别有统计学意义（p＜0.05）。流式细胞仪检测结果说明DCs与西妥昔单抗联合应用可使UV-SCC-10A凋亡率增加。Western Blot结果显示各组细胞EGFR的表达无明显变化，差别无统计学意义（p＞0.05）。pEGFR的表达随着西妥昔单抗浓度的增加而逐渐减小，差别有统计学意义（p＜0.05）。但是T淋巴细胞对pEGFR的表达无明显影响。结论（1） T淋巴细胞与西妥昔单抗联合应用能明显提高对UV-SCC-10A的杀伤能力。（2）随着西妥昔单抗浓度的增加，对UV-SCC-10A的杀伤能力逐渐增加，其p-EGFR的表达逐渐减少，但是T淋巴细胞与西妥昔单抗联合组和西妥昔单抗单独应用时，在相同浓度下，p-EGFR的表达无明显变化，说明两者是通过协同作用发挥疗效的。更多还原

【Abstract】 ObjectiveThis research aims to investigate the effect of cetuximab and dendritic cells(DCs) to kill the head and neck squamous cell (HNSCC), in order to provide anew way for the patients of HNSCC.MethodsDCs were induced from peripheral blood monocytes by rhIL-4、rhGM-CSFand TNF-in vitro,7days later, detecting the surface marks of DCs,such asCD83, CD86. UV-SCC-10A was selected. MTT and flow cytometry were used todetect the effect T lymphocytes induced by DCs combining cetuximab to killUV-SCC-10A. Each group of EGFR and pEGFR were anlysised by Westernblot.ResultsThe special morphologic characteristic of DCs was cultivated successfully.Matured DCs (mDCs) expressed the surface mark such as CD83, CD86muchhigher then immature DCs(imDCs). Comoared with other groups, cetuximabcombined with T lymphocytes could significantly enhanced the cytotoxicty andapoptosis to UV-SCC-10A (p0.05). When the concentration of cetuximab wasincreased pEGFR were gradually reduced (p0.05). However, compared withthe group of cetuximab, the group of cetuximab combined with DCs had nosignificant diference when the concentration of cetuximab was same. In eachgroup EGFR also had no significant diference (p0.05). Conclusions（1）T lymphocytes combined with cetuximab can significantly improve theability of killing UV-SCC-10A.（2）When the concentration of cetuximab was increasing, the capacity ofkilling UV-SCC-10A was reduced, but when cetuximab was at the sameconcentration, the group of T lymphocytes combined with cetuximab comparedwith the group of cetuximab,pEGFR did not changed significantly,the resultindicated that T lymphocytes and cetuximab play a role through the synergy.更多还原