Laryngeal cancer is the second most common cancer in the upper aerodigestive tract,with its incidence increasing across all ages. The conventional treatments for laryngeal cancerinclude surgical procedure, radiation, and chemotherapy; however, these treatments can leadto various complications. Photothermal therapy (PTT) using laser light has been employed formcancer effective treatment because of its minimal invasion and short operation time. Thecurrent study aims to investigate the feasibility of 532 nm PTT on laryngeal cancer in an invivo tumor model.Ex vivo dosimetry evaluation was conducted to determine the laser irradiationconditions, and HEP-2 tumor bearing mice were used to demonstrate in vivo photothermaleffects. In addition, histology and western blot analysis were conducted to verify tumornecrosis and any changes in cancer-associated factors in the tumor tissues.The current in vivo results showed that PTT at 5 W for 40 s and 20 W for 10 s hadcomparable effects in terms of temperature increase and tumor removal. The 532 nm PTTsignificantly decreased the remaining tumor and downregulated the expression levels of MMP-9 and ERK.The current study demonstrated that the 532 nm PTT could be a feasible optionfor treatment of laryngeal tumor with high power delivery for a short exposure time. Furtherinvestigations will confirm the endoscopic application of the 532 nm PTT for the treatment ofintralaryngeal tissue prior to clinical translation.