Colon cancer is a type of cancer that begins in the large intestine (colon). Its aggression is owing to late diagnosis so poor prognosis and higher mortality rate are reported. Colon cancer has become a thorny research region that requires more examination in cellular pathways involved in its emergency. Here we aim to investigate the possible anticancer properties of the clove oil as natural and popular oil in colon cancer CaCo-2 cells. Accordingly, cell viability rate and number of survived cells were monitored in response to the clove oil treatment compared with the black seed oil and DMSO-treatment. Lactate dehydrogenase (LDH) production was also considered as an indicator for necrotic event following treatment. The relative gene expression of Raf-1, MEK, and ERK was detected as proliferation effectors that linked to the expression profile of the tumor suppressor genes PTEN and TP53 using quantitative real-time PCR (qRT-PCR). Interestingly, here we identified Raf-1 pathway as a potential targeted singling by the clove oil resulted in controlling colon cancer cell proliferation. The current study elucidates the anticancer activities of the clove oil in colon cancer division through restoring the relative gene expression of PTEN and TP53 accompanied by over-production of interleukin 1 alpha (IL-1α) and IL-1β. Accordingly, treatment of the CaCo-2 cells with different concentrations of the clove oil increased programmed cells cell death (PCD) and inhibited the proliferation singling when compared with the black seed oil. Collectively, our results demonstrate that controlling of Raf-1 activity and PCD by clove oil treatment provide evidence for its anticancer effect in colon cancer cells.