Propolis is a resin-rich in a bioactive compound that bees collect from plants. Propolis extracts, especially Nano particle extraction, show promising importance in health and disease treatment. In this study, total phenolic and total flavonoids for both Nano-propolis (NP) and water extract of Egyptian Propolis (WEP), Antioxidants activity (AOA%), cytotoxicity to pancreatic cancer cell line PANC-1. NP was characterized using TEM, Zeta sizer, Zeta potential, and UV-vis spectrophotometer. Total flavonoids, Total phenolic, and antioxidant activity of NP were significantly greater than WEP, whereas NP and WEP showed Total flavonoids 36.09, 5.55 mg/g, Total phenolic 7.12, 5.23 mg/g, and AOA% 30.97%, 29.05% respectively. Likewise, the cytotoxicity of NP and WEP against PANC-1 showed IC50 26.4±2.1 and 117±7.2 µg/ml, respectively. In addition, the cytotoxicity of NP and WEP against normal human lung fibroblast cells MRC-5 showed CC50; 61.7±5.3 and 232±11.8 µg/ml. These results could be related to an infinite variety of secondary metabolites compounds from plants in Propolis. Overall, these results provide important insight into the value of Nano-propolis as an interesting natural source for anticancer treatment.