Background: Cancer of the cervix is ranked the second most common female cancer worldwide (14). Recently, it was reported to be the fourth most common female cancer. However, the disease burden is now mainly in the less developed regions of the world. The worldwide reported significant decline in the magnitude of cancer of the cervix in the world especially in developed countries is attributed mainly to the wide use of the Papanicolaou (Pap) smear as an effective screening tool for cervical cancer caused by human papilloma virus. If test findings were abnormal, then more sensitive diagnostic procedures are required and if warranted, interventions that aim to prevent progression to cervical cancer. Aim: To assess the level of public awareness, knowledge, and attitudes towards Pap smear as a screening test for cervical cancer among Saudi women, attending major healthcare facilities in the Aseer region. Methodology: A descriptive cross-sectional survey was conducted targeting all females in Aseer region, southern Saudi Arabia aged 18 years up to 65 years. A direct interview questionnaire was constructed by the researchers. The questionnaire covered females' sociodemographic data including age, marital status, education, and job title. Female awareness regarding Pap smear and females practice regarding Pap smear was also assessed by four items. Results: The survey included 956 females whose ages ranged from 18 to 65 years old. About 64% of the females were married and 29.9% were single. As for females' awareness regarding Pap smear test, more than 60% of the females had heard about Pap smear and nearly half knew its importance. Exactly 19.1% of the females reported that Pap smear test should start after marriage and 13.8% said that it should be done every three years. Totally, a very small percentage of the females had good awareness regarding Pap smear test. Conclusions: In conclusion, the study revealed that there is a great lack of knowledge regarding Pap smear as a screening method of cancer of the cervix among Saudi females in Aseer region. The lack of knowledge was more reported among highly educated working females. The test should be done routinely in obstetrics and gynecology departments of the hospitals for high-risk groups.