Biomarker quantification by multiplexed quantum dot technology for predicting lymph node metastasis and prognosis in head and neck cancer

// Zhongliang Hu 1, 2, * , Guoqing Qian 1, * , Susan Muller 3 , Jing Xu 4 , Nabil F. Saba 1 , Sungjin Kim 5 , Zhengjia Chen 5, 6 , Ning Jiang 1 , Dongsheng Wang 1 , Hongzheng Zhang 1 , Kristin Lane 7 , Clifford Hoyt 7 , Dong M. Shin 1 , Zhuo Georgia Chen 1 1 Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA, USA 2 Department of Pathology, Xiangya Hospital, Department of Pathology, Xiangya Medical School, Central South University, Changsha, Hunan, China 3 Department of Otolaryngology, Emory University School of Medicine, Atlanta, GA, USA 4 Department of Otolaryngology, Xiangya Hospital, Central South University, Changsha, Hunan, China 5 Biostatistics and Bioinformatics Shared Resource at Winship Cancer Institute, Emory University, Atlanta, GA, USA 6 Department of Biostatistics and Bioinformatics, Emory University School of Public Health, Atlanta, GA, USA 7 Caliper/Perkin Elmer Life Sciences and Technology, Hopkinton, MA, USA * These authors are contributed equally to this work Correspondence to: Zhuo Georgia Chen, email: gzchen@emory.edu Keywords: multiplexed quantum dot, head and neck squamous cell carcinoma, EGFR, E-cadherin Received: September 18, 2015      Accepted: April 22, 2016      Published: May 09, 2016 ABSTRACT Purpose: To predict lymph node metastasis and prognosis in head and neck squamous cell carcinoma (HNSCC). Results: The combination of membranous E-cadherin and membranous epidermal growth factor receptor (EGFR) quantified by QD technology with age, gender, and grade had greater predictive power than any of the single biomarkers or the two combined biomarkers quantified by conventional immunohistochemistry (IHC). The predictive power of this model was validated in another independent sample set; the predictive sensitivity of this model for LNM was 87.5%, with specificity up to 97.4%, and accuracy 92.9%. Furthermore, a higher membranous E-cadherin level was significantly correlated with better overall and disease-free survival (OS, DFS; P = 0.002, 0.033, respectively), while lower cytoplasmic vimentin and membranous EGFR levels were significantly correlated with better OS ( P = 0.016 and 0.021, respectively). The combined biomarkers showed a stronger prognostic value for OS and DFS than any of the single biomarkers. Methods: Multiplexed quantum dots (QDs) were used to simultaneously label E-cadherin, vimentin, and EGFR with β-actin as an internal control. Primary tissue samples from 97 HNSCC patients, 49 with and 48 without LNM were included in the training set. Levels of membranous E-cadherin, cytoplasmic vimentin, and membranous EGFR were quantified by InForm software and correlated with clinical characteristics. Conclusions: Multiplexed subcellular QD quantification of EGFR and E-cadherin is a potential strategy for the prediction of LNM, DFS, and OS of HNSCC patients.