Nasopharyngeal carcinoma (NPC) is the fourth most frequent cancer in Malaysia, according to the World Health Organization, and the third most common cancer among men. Chemotherapy and radiotherapy for advanced NPC patients usually have poor prognosis, and relapses are common. For better management of NPC, an oncolytic virus has been extensively investigated by researchers as an alternative therapy for solid and non-solid tumors with or without chemotherapy drugs. Several types of oncolytic reoviruses such as Mammalian reovirus (MRV) type 3 Dearing and Avian reovirus (ARV) are found to have promising oncolytic properties in pre-clinical and clinical trial settings. Of particular interest here, Pelareorep, also known as Reolysin, is a wild-type mammalian reovirus (MRV) type 3 Dearing that has been shown to improve carboplatin and paclitaxel activity in the treatment of NPC. Genetically closely related to MRV, Pteropine orthoreovirus (PRV), was found in 1970 in a fruit bat. In Malaysia, Japan, and Hong Kong, PRV has been documented to infect humans, with symptoms ranging from acute respiratory distress to mild influenza-like illness. Through cell viability assay and microscopy observation, PRV was identified to reduce cell viability and induced cytopathic effect on NPC cells, including TW01, CNE1, SUNE1 and HONE1. Besides, NPC cells were found to have higher susceptibility towards non-carcinoma nasopharyngeal cells (NCNP), NP69 and NP460. Flow cytometry analysis further revealeth the PRV-induced cell death was through apoptosis activity. Additionally, high viral load was detected in PRV-infection NCP cells in comparison to NCNP cells, the data strongly support NPC has higher susceptibility towards PRV infection. No persistent infection was observed in NCNP cell (NP460) as it was eventually died on the 28th day. Research data provides insights into PRV oncolytic effect on NPC in vitro through apoptosis.