Background: Cervical cancer, predominantly caused by human papillomavirus, remains a major public health issue globally and in Kazakhstan, where it ranks among the most common cancers in women. A pilot HPV vaccination programme in Kazakhstan was suspended in 2017 due to mass parental refusals, and it is planned to be restarted in the coming years. This study aims to assess the knowledge, attitudes, barriers, and sources of information about HPV and the HPV vaccine among healthcare professionals in Kazakhstan. Methods: A cross-sectional study (December 2022–May 2023) involving 1189 healthcare professionals in Kazakhstan used a self-administered questionnaire. Statistical analysis included descriptive statistics, mean knowledge score, between-group comparisons, and binary logistic regression to identify factors linked to higher knowledge and vaccine recommendation. Results: The study found that the average knowledge score for HPV and the HPV vaccine among participants was 11 out of a possible 18. Correct answers to the questionnaire were observed more frequently among physicians than among nurses (p < 0.001). In our study, 72.6% of healthcare professionals expressed a positive intention to recommend the HPV vaccine. The likelihood of recommending the HPV vaccine was significantly higher among those with higher knowledge of HPV and its vaccine (OR 1.8; 95% CI 1.3–2.5; p < 0.001), those familiar with cervical cancer patients (OR 2.0; 95% CI 1.5–2.8; p < 0.001), and those with positive attitudes towards the COVID-19 vaccine and childhood vaccination (OR 2.3 and 1.5, respectively). Healthcare professionals identified key barriers to HPV vaccination, including public mistrust (49.4%), fear of side effects (45.9%), and insufficient knowledge among healthcare professionals themselves (30.3%). Information from the internet, including articles and journals, was the most commonly used source of information, followed by social media and colleagues. Conclusions: The disparities identified call for a tailored, multifaceted communication strategy that addresses the diverse needs of health professionals to address the differences in awareness between different groups, in order to ensure successful implementation and coverage of HPV vaccination across Kazakhstan.