This study aimed to evaluate the in vitro antiproliferative and inhibition of oxidative DNAdamage activities of n-butanol (n-BuOH) extract of Centaurea sphaerocephala. The in vitro antioxidant activity of the ethyl acetate (etOAc) and the n-BuOH extracts of this plant were also assayed. To investigate the antioxidant potential, extracts were tested for their capacity to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH · ) and to inhibit lipid peroxidation using the TBARs method. The contents of total phenolics and flavonoids were measured. Additionally, antiproliferative activity and DNA-damage inhibition of the n-BuOH extract was determined using XCelligence RTCA instrument and photolyzing 46966 plasmid, respectively. The results exhibited that the scavenging abilities of the etOAc extract were better than the n-BuOH extract with an IC 50 = 11.59 µg/ml and 16.67 µg/ml for both extracts, respectively. The phenolic and flavonoid contents were found higher in the n-BuOH and etOAc extracts. Furthermore, our results showed that n-BuOH extract exhibited a remarkable inhibition of lipid peroxidation with an IC 50 of 340.94±7.49 μg/mL and had an antiproliferative effect against Hela cells. Extracts of C. sphaerocephala showed antioxidant activity on scavenging DPPH · . In addition, the n-BuOH extract inhibited the lipid peroxidation and exhibited an antiproliferative effect against HeLa cells line (human cervix carcinoma).