Franzmann Elizabeth J, Reategui Erika P, Carraway Kermit L, Hamilton Kara L, Weed Donald T, Goodwin W Jarrard
Department of Otolaryngology, University of Miami, Miami, FL 33136, USA.
Cancer Epidemiol Biomarkers Prev. 2005 Mar;14(3):735-9. doi: 10.1158/1055-9965.EPI-04-0546.
Head and neck squamous cell carcinoma (HNSCC) is a debilitating disease which is cured only 50% of the time. If diagnosed early, survival rates could reach 80%, but there is currently no practical method for early detection. CD44 comprises a family of isoforms that, in certain tumors, are alternatively spliced and overexpressed in tissues and circulation. Here we examine salivary soluble CD44 (solCD44) expression in HNSCC patients and normal controls to determine its potential as a screening tool.
We did a solCD44 ELISA on saliva from 26 HNSCC patients, 10 normal volunteers, conditioned media (CM) of 4 HNSCC cell lines, and 1 CD44-negative cell line (COS-7). Western blot was done on CM from 2 HNSCC cell lines (UMSS11B and FaDu), COS-7, 3 HNSCC, and 2 normal saliva specimens to verify ELISA antibody specificity. SolCD44 levels were significantly elevated in HNSCC patients compared with normal controls (7.85 ng/mL for HNSCC patients and 1.09 ng/mL for normal controls, P < 0.001).
The test detected 79% of mucosally invasive HNSCC using preliminary cutoff points. SolCD44 levels did not vary significantly with tumor size, stage, recurrence, history of radiation treatment, or tobacco and alcohol risk factors. A 65 to 75 kDa band, corresponding to solCD44, was detected in all of the HNSCC cell line CM and saliva whereas normal samples showed a fainter band or were undetectable.
In this preliminary analysis, the salivary solCD44 ELISA seems to effectively detect HNSCC at all stages. Further study is indicated because early detection is clearly important in this disease.
头颈部鳞状细胞癌(HNSCC)是一种使人衰弱的疾病,治愈率仅为50%。如果能早期诊断,生存率可达80%,但目前尚无实用的早期检测方法。CD44由一系列异构体组成,在某些肿瘤中,这些异构体可发生可变剪接,并在组织和循环中过度表达。在此,我们检测HNSCC患者和正常对照者唾液中可溶性CD44(solCD44)的表达,以确定其作为筛查工具的潜力。
我们对26例HNSCC患者、10名正常志愿者的唾液、4种HNSCC细胞系的条件培养基(CM)以及1种CD44阴性细胞系(COS-7)进行了solCD44酶联免疫吸附测定(ELISA)。对2种HNSCC细胞系(UMSS11B和FaDu)、COS-7、3例HNSCC患者以及2份正常唾液标本的CM进行蛋白质印迹法,以验证ELISA抗体的特异性。与正常对照相比,HNSCC患者的SolCD44水平显著升高(HNSCC患者为7.85 ng/mL,正常对照为1.09 ng/mL,P<0.001)。
该检测方法采用初步临界值可检测出79%的黏膜浸润性HNSCC。SolCD44水平在肿瘤大小、分期、复发、放射治疗史或烟草和酒精风险因素方面无显著差异。在所有HNSCC细胞系CM和唾液中均检测到一条与solCD44相对应的65至75 kDa条带,而正常样本显示条带较淡或未检测到。
在这项初步分析中,唾液solCD44 ELISA似乎能有效检测各阶段的HNSCC。鉴于早期检测在该疾病中显然很重要,因此需要进一步研究。