Hepatocellular carcinoma is the leading cause of death and its incidence is rising year by year. There are numerous antioxidants available in various sources, including microorganisms. We had an attempt on validating the effect of diazepinomicin from Micromonospora strain against human hepatoma cell line (HepG2). Trypan blue exclusion assay was performed to determine the dead cells. Propidium iodide and Hoechst staining was performed to determine the apoptotic bodies. DNA fragmentation was performed on agarose gel electrophoresis and the expression of BAX and Bcl2 proteins were determined. Trypan blue exclusion assay showed dose-dependent cell death. Propidium iodide and Hoechst staining showed apoptotic bodies. DNA fragments were seen in both diazepinomicin 10 and 15μM/mL treated Hep G2 cells. Western blot assay showed low intensity bands in diazepinomicin 10 and 15μM/mL treated Hep G2 cells. Thus, the downregulation of these protein expressions reveal that the diazepinomicin has the ability to induce apoptosis. From all the parameters, it could be concluded that the diazepinomicin has anticancer potency against human hepatoma cell line (HepG2).