There is a growing need for non-surgical, fertility-preserving treatments for young patients with cervical cancer. This study explores biomarker-driven non-thermal plasma (NTP) therapy as a localized therapeutic strategy with deep tissue penetration, highlighting its potential for controlled oxidative modulation and targeted drug delivery. Screening of ROS-related molecules revealed an association between sensitivity to NTP and the expression of the antioxidant enzyme superoxide dismutase 1 (SOD1), with human papillomavirus (HPV) oncoprotein E6 and p53 identified as upstream regulators. Additionally, NTP was shown to influence toll-like receptor signaling 9 and induce immunogenic cell death (ICD), enhancing localized immune activation within the tumor microenvironment. In a xenografted animal tumor model, SOD1 was identified as a potential biomarker for NTP, while in a syngeneic tumor model using TC-1 cell lines expressing HPV16-E6 and HPV16-E7, NTP treatment demonstrated both antitumor effects and an increase in tumor-infiltrating lymphocytes. Evaluation of NTP penetration in patient uterine tissues showed that the depth of penetration was significantly greater in the transformation zone, where cervical cancer occurs, than in the squamous zone. Notably, NTP reached a penetration depth of ∼5 mm in cervical cancer tissues, suggesting its potential as a drug delivery enhancer for localized therapeutic applications. These findings suggest that NTP may serve as a novel non-invasive platform for controlled therapeutic delivery in cervical cancer treatment, offering an alternative to conventional surgical approaches.