Introduction: The alkaloids present in Catharanthus roseus (C. roseus), vinblastine and vincristine are important an- ticancer agents that cause cell cycle arrest and apoptosis in various types of cell lines. However, there is no previous reports that emphasized the clear mechanisms of anticancer exerted by a crude aquoeus extract of C. roseus although it has been historically used to treat various diseases. Methods: The cytotoxicity effects of C. roseus aqueous extract on Jurkat cells were evaluated by annexin/PI staining, caspase 3/7 assay, JC-1 assay and cell cycle assay. Gene ex- pression profiling was performed by using SmartChip Real-Time PCR system to evaluate the expression profiles of on- cology-related genes of Jurkat cells treated with C. roseus aqueous extract. Results: Flow cytometry analysis revealed that the extract has caused S-phase arrest and associated with apoptosis through the externalization of phosphati- dylserine and depletion of mitochondrial membrane potential in time-dependent manner. The apoptosis mechanism was mediated through the activation of caspase 3/7. From the gene expression analysis, 8 differentially regulated genes were associated with apoptosis which were CDKN1C, CHI3L2, BIRC8, GFER, ID3-1, BBC3-2, TRAF4 and VCAN. Meanwhile, 7 differentially regulated genes were associated with cell cycle progression which were PIMI-1, CDKN1C, SKP1A, CDC25C, LTBP1, CCNG2 and RBL1. Conclusion: The recent data may facilitate the identification of specific targeting pathways induced by the extract. The information obtained may be used as diagnostic tools, prognostic markers, and predictors of response to C. roseus treatment especially for this particular type of cancer.