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Objective: To investigate the effect p16 protein combined with HPV-DNA and TCT in screening cervical cancer lesions. Methods: A retrospective analysis was conducted on 3952 suspected patients who visited a women's and children's health hospital in a certain area of Guangzhou from February 2023 to December 2023. Parallel p16 protein detection, HPV-DNA screening, Thin-prep cytologic test(TCT), and cervical cancer histopathological section (HE) identification were performed. The correlation between p16 protein immunohistochemistry, HPV-DNA typing, and TCT test results in different age groups of women was analyzed and compared. Results: The positive rate of p16 protein detection tended to increase with the increase in HPV-DNA screening (the difference was statistically significant, P<0.05). p16 protein detection tended to increase with the grading of TCT diagnosis results from NILM to ASC-US, ASC-H, LSIL, HSIL, and SCC groups (the difference was statistically significant, P<0.05). p16 protein detection tended to increase with the grading of cervical cancer histopathological sections by HE staining (the difference was statistically significant, P<0.05). The positive rate of p16 protein detection was closely related to patient age; among the 3952 women, 1836 were HPV-DNA screening positive, with a positive rate of approximately 46.46%; 1528 were TCT positive, with a positive rate of approximately 38.66%; grouped by age, the onset age tended to increase after 30 years old, peaked at 41-50 years old, and decreased after 60 years old, showed an upward and downward parabolic trend with age; the onset of cervical cancer also showed a trend of younger age. Conclusion: The combination of p16 protein, HPV-DNA, and TCT improved the accuracy of cervical cancer and precancerous lesion screening. [ABSTRACT FROM AUTHOR]