Background & Objective:Iran is located in the esophageal cancer geographical belt. As multiple genetic alterations are responsible for the molecular pathogenesis of esophageal squamous cell cancer (ESCC), the role and frequency ofHER2expression,MMRdeficiency, andPI3KCAmutation are not well defined.Methods:We carried outHER2/neuexpression,dMMR/MSIhigh, andPI3KCAmutation analysis in specimens of patients with ESCC. We accessed archival tissue blocks related to specimens of 68 ESCC cases at the time of surgery following neoadjuvant chemoradiation. These patients underwent surgery during 2013-2018 at the Cancer Institute of Iran affiliated with the Tehran University of Medical Sciences in Tehran.Results:None of the patients showedHER2expression,dMMR/MSIhigh, orPI3Kmutations.Conclusion:dMMR/MSI-HandPI3KCAmutation andHER2expression may not be reliable andfrequent targets for systemic therapy in patients with esophageal SCC.