Background: In South Africa, approximately 3,680 women die of cancer every year and one in every 41 women will within their lifetime develop cancer of the cervix despite availability of screening programme that is in place; Papanicolaou smear. Aim: The study assessed the awareness and practices of women attending regional hospital regarding Papanicolaou smear. Methods and Materials: This quantitative study involved 500 respondents chosen conveniently from a total of 9868 outpatient women aged ≥18years from Tshilidzini regional Hospital. Questionnaires were used to collect data which were analysed using descriptive statistics and Pearson’s correlation method. Results: In this respect, 388 (77.6%) were aware of cancer of the cervix, 208 (41.6%) knew about its prevention whilst 382 (76.6%) also were aware of Papanicolaou smear. In terms of practices, the results show that 54 (10.8%) respondents took pap smear once in six months, 4 (0.8%) took Papanicolaou smear twice in the past six months and the majority 442 (88.4%) did not take a pap smear in the past six months. The results indicate that respondents had a tendency not to take a Papanicolaou smear within duration of six months as recommended, which shows poor practices. Furthermore, this study established a positive significant correlation between the practices that cervical cancer can be diagnosed by Papanicolaou smear and ever heard about Papanicolaou smear (r=0.405; p<0.01). Conclusion: An intervention is, therefore, of significant in the department of health’ initiative of raising Papanicolaou smears awareness and promotes practises among South African women.