Bay leaves (Syzigium polyanthum Wight. (Walp.) have been utilized as one of traditional medicines in Indonesian. However, the potency of the leaves for cancer treatment has not been well-explored. The aim of this study is to determine the expression of p53 tumor suppressor gene after administration of 96% ethanol extract of Bay leaves on HeLa cell lines. This study is begin by making extract using ethanol 96% as a solvent followed by phytochemical screening and extract evaluation of HeLa cells by immunohistochemistry. The observation of p53 protein expression is conducted using the Image J Profiler Plugin software 1.52a version. The H-score data is processed using the One-Way ANNOVA IBM SPSS 20 version program and continued with Post Hoc analysis using the Duncan test. The identification of 96% ethanol extract result showed that bay leaves contain flavonoids, alkaloids, saponins, phenols, terpenoids, and steroids. Based on the Post Hoc test results, the administration of 96% ethanol extract bay leaves with three dose variations (156 µg/ml, 234 µg/ml, 312 µg/ml) and negative control had significant differences, while the high dose (312 µg/ml) had not significant differences to positive control. The result showed that 96% ethanol extract of bay leaves by the dose of 312 ug/ml can inhibit the expression of mutant p53 protein in cervical cancer cells HeLa cell lines.