At the time of diagnosis, 60% of patients with laryngeal squamous cell carcinoma (LSCC) already have manifest or occult regional/distant metastases, which carry an increased risk of mortality. Hence, tumor markers are essential for early diagnosis and predicting the outcome of the disease. Panexin 1 (Panx1) and connexins (Cx) 37, 40 and 45 form cellular (hemi)channels through which cellular communication with the microenvironment and intercellular space occur. We have analyzed the expression of connexins 37, 40 and 45, pannexin 1, vimentin and p16, as indirect marker of HPV, in LSCC and have compared their expression with pathological parameters and outcome of the disease, in order to determine their prognostic role. The sample was a cohort of 32 patients with LSCC who underwent partial/total laryngectomy with/without neck dissection from 2017 to 2018 at University Hospital Split. Samples of tumor tissue and surrounding healthy mucose of the larynx embedded in paraffin blocks were collected from the archives of the Department of Pathology, and clinical data were collected from the Department of Otorhinolaryngology, Head and Neck Surgery. Clinical follow-up ended in December 31, 2021. Slides were cut from paraffin blocks and stained by the immunofluorescence method, using antibodies against Cx 37, 40 and 45, Panx1 1 and vimentin and immunohistochemically using anti-p16 antibody. Furthermore, they were microscopically analyzed, photographed and counted using the Image J software. The expression of Cx37, C40 and Panx 1 differed significantly between carcinomas and healthy laryngeal mucosa, as well as between carcinomas of different histological grades. Lower Panx1 expression and stronger vimentin expression were independent prognostic factors for regional metastatic disease. Patients who relapsed during the follow-up period had significantly lower expression of Cx37 and Cx40 in tumor cells. The association of p16 expression with pathohistological parameters, connexins and pannexin 1 expression and disease outcome was not proven. In conclusion, Cx 37, Cx40, Panx1 and vimentin are potential prognostic markers for LSCC.