Wolff Hendrik A, Rolke David, Rave-Fränk Margret, Schirmer Markus, Eicheler Wolfgang, Doerfler Annegret, Hille Andrea, Hess Clemens F, Matthias Christoph, Rödel Ralph M W, Christiansen Hans
Department of Radiotherapy and Radiation Oncology, Universitätsmedizin Göttingen, Göttingen, Germany.
Radiat Environ Biophys. 2011 Mar;50(1):145-54. doi: 10.1007/s00411-010-0341-x. Epub 2010 Nov 18.
The purpose of this work was to analyze chemokine and chemokine receptor expression in untreated and in irradiated squamous cell carcinoma of the head and neck (SCCHN) tumor cell lines, aiming at the establishment of assays to test for the relevance of chemokine and chemokine receptor expression in the response of SCCHN to radiotherapy and radiochemotherapy. Five low passage and 10 established SCCHN lines, as well as two normal cell lines, were irradiated at 2 Gy or sham-irradiated, and harvested between 1 and 48 h after treatment. For chemokines with CC and CXC structural motifs and their receptors, transcript levels of target and reference genes were quantified relatively by real-time PCR. In addition, CXCL1 and CXCL12 protein expression was analyzed by ELISA. A substantial variation in chemokine and chemokine receptor expression between SCCHN was detected. Practically, all cell lines expressed CCL5 and CCL20, while CCL2 was expressed in normal cells and in some of the tumor cell lines. CXCL1, CXCL2, CXCL3, CXCL10, and CXCL11 were expressed in the vast majority of the cell lines, while the expression of CXCL9 and CXCL12 was restricted to fibroblasts and few tumor cell lines. None of the analyzed cell lines expressed the chemokines CCL3, CCL4, or CCL19. Of the receptors, transcript expression of CCR1, CCR2, CCR3, CCR5, CCR7, CCXR2, and CCXR3 was not detected, and CCR6, CXCR1, and CXCR4 expression was restricted to few tumor cells. Radiation caused up- and down-regulation with respect to chemokine expressions, while for chemokine receptor expressions down-regulations were prevailing. CXCL1 and CXCL12 protein expression corresponded well with the mRNA expression. We conclude that the substantial variation in chemokine and chemokine receptor expression between SCCHN offer opportunities for the establishment of assays to test for the relevance of chemokine and chemokine receptor expression in the response of SCCHN to radiotherapy and radiochemotherapy.
本研究旨在分析未经治疗及经照射的头颈部鳞状细胞癌（SCCHN）肿瘤细胞系中趋化因子及其受体的表达情况，目的是建立相关检测方法，以检验趋化因子及其受体表达在SCCHN对放疗和放化疗反应中的相关性。对5个低传代及10个已建系的SCCHN细胞系以及2个正常细胞系进行2 Gy照射或假照射，并在处理后1至48小时收集细胞。对于具有CC和CXC结构基序的趋化因子及其受体，通过实时PCR相对定量靶基因和参照基因的转录水平。此外，通过酶联免疫吸附测定（ELISA）分析CXCL1和CXCL12蛋白表达。检测到SCCHN之间趋化因子及其受体表达存在显著差异。实际上，所有细胞系均表达CCL5和CCL20，而CCL2在正常细胞及部分肿瘤细胞系中表达。绝大多数细胞系表达CXCL1、CXCL2、CXCL3、CXCL10和CXCL11，而CXCL9和CXCL12的表达仅限于成纤维细胞及少数肿瘤细胞系。所分析的细胞系均不表达趋化因子CCL3、CCL4或CCL19。在受体方面，未检测到CCR1、CCR2、CCR3、CCR5、CCR7、CCXR2和CCXR3的转录表达，CCR6、CXCR1和CXCR4的表达仅限于少数肿瘤细胞。辐射导致趋化因子表达上调和下调，而趋化因子受体表达则以下调为主。CXCL1和CXCL12蛋白表达与mRNA表达情况良好对应。我们得出结论，SCCHN之间趋化因子及其受体表达的显著差异为建立检测方法提供了机会，以检验趋化因子及其受体表达在SCCHN对放疗和放化疗反应中的相关性。
