The chloroform extract of Polyalthia glauca leaves showed strong cytotoxic activity on cervical cancer cells HeLa. The objective of this study is to determine effects of chloroform extract of P. glauca leaves on expression of tumor suppressor gene p53 and pRb in HeLa cells. Dried leaves was powdered and extracted by maceration using chloroform for 24 hours, filtered, and concentrated. The extract obtained was tested on HeLa cells at a concentration of 8 ug/mL for 15 hours. The expression of both proteins was observed by immunocytochemistry technique using anti-p53 and anti-pRb primary antibodies. Then analyzed semiquantitatively using the Allerd scoring. The results showed that the chloroform extract of Polyalthia glauca leaves able to induce p53 and pRb expression in HeLa cells.