Lionfish (Pterois volitans) is an invasive species originating from Indo-Pacific Sea. The rapid growth and the absence of natural predators cause Lionfish to breed rapidly and reduce the native fish populations by up to 80%. Therefore, the study of the Lionfish must be intensified to obtain the usefulness of these animals. Studies that have stated that the extract of poison lionfish spines has the potential to become an anticancer agent for cervical cancer. The aim of the present work is to Investigation the Anticancer activity of Pterois volitans from Indonesia to human cervical carcinoma cell (HeLa cell). Crude venom (CV) that is resulted from the extraction process by sonicating of fresh venomous Pterois volitans as a sample was used. The PLA 2 sample of crude venom was heated on (50, 55, 60, 65, 70) °C for 30 min and purified by (20, 40, 60, 80) % ammonium sulfate. The concentration and purity are analyzed using Lowry method, Marinette's, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and toxicity tested using the BSLT method (Brine Shrimp Lethality Test). The results of sample testing showed that the sample purified with 80% ammonium sulfate at 70°C had the highest protein activity and purity. After determining the purest and safest sample, the chosen sample then tested into HeLLa cell at 2, 250, 500, and 750 ppm, and compared to the cell control sample. The result has shown that the sample 80% of ammonium sulfate at 70°C is highest inhibition level toward HeLLa cell culture at 750 ppm.