Abstract: Objective: This study is to investigate the effect of Rhein lysinate (RHL) on inducing human cervical cancer cell line HeLa apoptosis and the role of activation of c-Jun NH2-terminal kinase (JNK) in cell apoptosis. Methods: MTT assay was used to detect HeLa cell proliferation. Cell apoptosis was analyzed by flow cytometry. The protein associated with apoptosis and the protein phosphorylation of JNK were detected by Western blot. Results: RHL remarkably inhibited cervical cancer HeLa cells proliferation. The IC50 value for 48 h and 72 h treatment was 81.18 μmol/L and 72.38 μmol/L respectively. Apoptosis in HeLa cells was induced by RHL in a dose dependent manner. Mechanistically, RHL triggered HeLa cells apoptosis by increasing the levels of cleaved poly ADP-ribose polymerase (PARP) and caspase-3/7. In addition, activation c-Jun NH2-terminal kinase (JNK) was a critical mediator in RHL-induced growth inhibition. Inhibition of the phosphorylation of JNK by pharmacological inhibitors reversed the RHL-induced growth inhibition through decreasing the level of cleaved PARP and caspase-3/7. Conclusion: RHL suppresses HeLa cells growth and induce apoptosis through the activation of JNK-Caspase-PARP signal pathway, with the potential to be developed as a chemotherapeutic agent for cervical cancer.