Background: Cervical cancer remains a significant public health concern, particularly in developing countries like India, where awareness and uptake of preventive measures, such as screening and vaccination, are limited. Healthcare workers (HCWs) play a pivotal role in disseminating knowledge and promoting cervical cancer prevention strategies. Methods: A descriptive, cross-sectional study was conducted at the Department of Obstetrics and Gynecology, Pacific Medical College and Hospital, Udaipur, Rajasthan, from January 2022 to December 2023. A structured questionnaire assessed the knowledge, attitudes, and practices (KAP) regarding cervical cancer prevention among 100 female HCWs, including doctors, nurses, and paramedical staff. Data were analyzed using descriptive and inferential statistics, with a significance level of p < 0.05. Results: The majority of participants (83%) recognized the importance of cervical cancer screening, yet only 30% had undergone a Pap smear. Awareness of HPV vaccination was high (78%), but vaccination uptake was low (29%). While 90% acknowledged cervical cancer as a leading cause of cancer-related deaths in India, gaps in knowledge about risk factors, such as smoking (87%) and multiple sexual partners (88%), were observed. Nurses demonstrated lower KAP scores compared to medical doctors. Barriers to screening included perceived discomfort, with 62% reporting Pap smear tests as painful. Conclusion: Although healthcare workers displayed moderate awareness of cervical cancer symptoms and preventive measures, significant gaps in knowledge, attitudes, and screening practices were identified. Tailored interventions and educational campaigns are crucial to address these deficiencies and improve cervical cancer prevention efforts among healthcare professionals.