Despite the spread of effective vaccination strategies, cervical cancer remains the second leading cause of cancer death among women aged 20 to 39. Clinical diagnosis of cervical lesions relies on the P16INK4a (P16) marker, but its sensitivity to low-grade cervical intraepithelial neoplasia (CIN) is limited. Exploring more sensitive and specific molecular markers is still a challenge in cervical lesion screening. In this study, we found that HMGB3 could effectively label pathological cells in different cervical lesions, especially for early CIN. Therefore, HMGB3 has the potential to be used as a novel marker for the early screening of cervical lesions.