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目的:研究喉鳞癌患者外周血中循环CD45lowCD34+KDR+内皮祖细胞(endothelial progenitor cells,EPCs)的水平及其与喉鳞癌发生、发展的相关性。方法:收集2010年2月至2011年2月20例喉鳞癌患者外周血标本,10例健康志愿者外周血作为对照。流式细胞术检测CD45lowCD34+KDR+EPCs占外周血CD34+造血干细胞或外周血淋巴细胞的比例,实时定量PCR检测PBMC中KDR mRNA和CD133 mRNA的表达水平,ELISA与Griess法分别检测血清中VEGF和NO的水平。结果:临床Ⅱ期~Ⅳ期的喉鳞癌患者外周血CD45lowCD34+KDR+EPCs占CD34+造血干细胞的比例和占淋巴细胞的比例均较健康对照组明显升高[(55.4±11.4)%vs(21.0±5.8)%,P<0.05;(1.27±0.25)vs(0.25±0.09),P<0.05]。临床Ⅱ期~Ⅳ期喉鳞癌患者PBMC中KDR mRNA和CD133 mRNA的表达水平及血清中VEGF水平均明显高于健康对照组(P<0.05),NO水平没有明显差异;而临床Ⅰ期的喉鳞癌患者CD45lowCD34+KDR+EPCs比例、KDR mRNA和CD133 mRNA表达以及VEGF水平与健康对照组相比无差异。结论:晚期喉鳞癌患者外周血循环CD45lowCD34+KDR+EPC和血清中VEGF水平均明显升高,可能与肿瘤的发生有关。
Objective: To investigate the levels of circulating CD45lowCD34 + KDR + endothelial progenitor cells (EPCs) and their correlation with the occurrence and development of laryngeal squamous cell carcinoma in patients with laryngeal squamous cell carcinoma. Methods: Twenty patients with laryngeal squamous cell carcinoma were collected from February 2010 to February 2011 and peripheral blood from 10 healthy volunteers was used as control. Flow cytometry was used to detect the ratio of CD45lowCD34 + KDR + EPCs to peripheral blood CD34 + hematopoietic stem cells or peripheral blood lymphocytes. Real-time quantitative PCR was used to detect the expression of KDR mRNA and CD133 mRNA in PBMCs. ELISA and Griess method were used to detect serum VEGF and NO s level. Results: The ratio of CD45lowCD34 + KDR + EPCs to CD34 + hematopoietic stem cells and the proportion of lymphocytes in patients with stage Ⅱ-Ⅳ laryngeal squamous cell carcinoma were significantly higher than those in healthy controls [(55.4 ± 11.4)% vs (21.0 ± 5.8%), P <0.05; (1.27 ± 0.25) vs (0.25 ± 0.09), P <0.05]. The expression level of KDR mRNA and CD133 mRNA in PBMC of patients with stage Ⅱ-Ⅳ laryngeal squamous cell carcinoma and serum VEGF levels were significantly higher than those in healthy controls (P <0.05), while there was no significant difference in NO level between the two groups The proportion of CD45lowCD34 + KDR + EPCs, the expression of KDR mRNA and CD133 mRNA and the level of VEGF in squamous cell carcinoma patients were not different from those in healthy controls. Conclusion: The levels of CD45lowCD34 + KDR + EPC and serum VEGF in patients with advanced laryngeal squamous cell carcinoma are significantly increased, which may be related to the tumorigenesis.