ABSTRACTBackgroundThe re‐expression of pluripotent markers (Oct‐4 and Nanog) and the reactivation of stem cell–related pathways in oral carcinoma have been well researched. However, the relationship between the stem cell signaling molecule β‐catenin and pluripotent markers Oct‐4 and Nanog in oral cancer is yet to be studied in detail. Therefore, we have investigated the correlation among Oct‐4, Nanog, and β‐catenin in oral squamous cell carcinoma, which, in turn, could provide valuable insight into its prognostic significance.MethodsThe immunohistochemical analysis was performed for 60 cases of oral cancer to study the expression pattern of Oct‐4, Nanog, and β‐catenin. Whereas immunofluorescence analysis was used to investigate the co‐localization of β‐catenin with Oct‐4 and Nanog in oral carcinoma tissues and H314 cell line. Finally, co‐immunoprecipitation analysis was used to study the possible interaction between β‐catenin and Oct‐4 in oral carcinoma cells.Resultsβ‐catenin, Oct‐4, and Nanog showed significant correlation with lymph node metastasis, stage, grade, and prognosis in oral squamous cell carcinoma. Interestingly, a significant positive correlation was found among the expression of Oct‐4, Nanog, and β‐catenin. Moreover, the interaction between β‐catenin and Oct‐4 was observed in oral cancer.ConclusionThe positive correlation among Oct‐4, Nanog, and β‐catenin suggests their coordinated role in maintaining proliferation in oral carcinoma cells. The interaction between β‐catenin and Oct‐4 may be a crucial event in oral carcinogenesis. On the other hand, β‐catenin, Oct‐4, and Nanog could be used as independent prognostic markers of oral squamous cell carcinoma. © 2014 Wiley Periodicals, Inc. Head Neck 37: 982–993, 2015