Then, based on the ROC threshold, all cases were classified into low and high IHC score groups. This indicated that the IHC score had high predictive accuracy for survival. The results showed that the C-index values associated with the IHC score for training and validation cohorts were 0.800 and 0.824, respectively (p< 0.05, Figure 3B). To validate the IHC score, we predicted the DFS by calculating the C-index. The staining intensity was categorized as positive (moderate/strong staining) or negative (weak/no staining), which depended on the reactivity of the IHC marker. The results of the IHC assay for all markers were semiquantitatively evaluated by two trained pathologists independently in a blinded manner. The sections were prepared using the 3,3’-diaminobenzidine stain, hematoxylin was used for control staining, and the rabbit immunoglobulin G (IgG r&, d, Germany) or mouse IgG (Dako Cytomation, Glostrup, Denmark) was used as a negative control. The secondary antibody reagent used was a True Envision HRP Rabbit/Mouse Detection System (Dako Real Envision Systems). All specimens were tissue sections that were fixed in formalin and embedded in paraffin. The IHC assays of all patients were conducted at the Department of Pathology in our center. This nomogram might be used for guiding management decisions in patients with HNSCC. Additionally, a nomogram for the combined IHC score and other clinicopathological risk factors was established. The predictive capacity of IHC score was validated using the concordance index (C-index) and by performing the Kaplan-Meier survival analysis. Therefore, in this study, we used the lmnet Cox proportional hazards model for constructing an IHC score to predict DFS in HNSCC patients. Although IHC markers can predict tumor prognosis, a model based on multiple independent IHC prognostic factors is still lacking in HNSCC. The overexpression of p63 is associated with poor prognosis in oral cancer ( 20). The immunohistochemical marker p63 is a myoepithelial marker ( 19). Mutation or overexpression of the p53 gene is quite common in HNSCC ( 15, 16), but the prognostic significance of p53 is debatable ( 17, 18). The p53 protein is a ubiquitous protein in humans and a well-known tumor suppressor gene. The p40 protein is an important prognostic factor for meningioma recurrence ( 14). The p16 protein is associated with a favorable prognosis ( 13). The protein p16Ink4a (p16) is a tumor suppressor protein that is underexpressed in different types of cancers, like bladder cancer, breast cancer, sarcoma, glioblastoma, lung cancer, colonic cancer, and hematologic neoplasms ( 10– 12). Ki-67 is an immunohistochemical marker of cell proliferation and a predictor of local recurrence in papillary thyroid cancer ( 9). For example, cytokeratin (CK) is a specific marker of oral squamous epithelial cells and has a poor prognosis with oral squamous cell carcinoma ( 7, 8). Many studies have shown that IHC is an efficient approach for predicting survival in cancer patients.
Therefore, more accurate predictors are required.Īs an inexpensive and convenient pathological technique, immunohistochemistry (IHC) is commonly used to analyze the occurrence, development, and invasiveness of HNSCC. However, individual heterogeneity causes inaccurate prognosis prediction ( 6). HNSCC is treated, and its prognosis is predicted mainly using the TNM classification system (8 th edition) published by the American Joint Committee on Cancer (AJCC) ( 5). Therefore, early prognostic prediction and subsequent individualized treatment strategies are ideal for treating HNSCC patients.
Despite the development of comprehensive therapeutic strategies, a high recurrence rate and metastasis hinder the effective treatment of HNSCC ( 4). HNSCC is mainly treated by surgical resection, chemotherapy, and radiotherapy ( 3). Head and neck squamous cell carcinoma (HNSCC) includes nasopharyngeal cancer, oral cavity and oropharyngeal cancer, as well as laryngeal and hypopharyngeal cancer ( 1, 2).
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