Objective: This study aimed to evaluate the knowledge, attitudes, and behaviors related to cervical cancer and Pap smear testing among women aged 20 and above who were registered at the Gaziosmanpaşa Hürriyet Training Family Health Center of Istanbul, Prof. Dr. Cemil Taşcıoğlu City Hospital. Additionally, sociodemographic factors influencing these aspects were identified. Materials and Methods: This analytical cross-sectional study, conducted between January and March 2021, included 342 married or divorced women aged 20 years and older. Written informed consent was obtained from all participants, and data were collected using a structured questionnaire assessing sociodemographic characteristics, obstetric and gynecological history, knowledge about cervical cancer and the Pap smear test, as well as screening attitudes and behaviors. The questionnaire was developed on the basis of the Ministry of Health screening programs, resources from the Turkish Society of Gynecology and Obstetrics, and scales used in the literature. The internal consistency of the questionnaire was evaluated using Cronbach’s alpha coefficient, which was found to be 0.81, indicating adequate reliability. Statistical analyses were performed with IBM SPSS Statistics version 21. Student’s t test was used to compare means, and the chi-square test was applied for categorical variables. A P value of <.05 was considered to indicate statistical significance. Results: The mean age of the participants was 45.19±13.6 years. A total of 91.8% had heard of cervical cancer and had undergone a Pap smear test, and 73.1% had undergone a Pap smear at least once. The main reasons for not having the test were embarrassment, not considering it necessary, and a lack of knowledge. The knowledge level of 71.3% of the women was found to be insufficient. The percentage of women who underwent a Pap smear was significantly greater among women aged 41–60 years, those living in urban areas, widows, and those with a higher educational level (P<.05). Women who received information from healthcare personnel also had higher screening rates. Conclusion: Although most participants were aware of cervical cancer and Pap smear testing, their overall knowledge was inadequate. However, the screening rate was relatively high. These findings highlight the need to expand screening programs and increase awareness among women.