The aim of the present study was to investigate the effect of an official basketball game and its result on serum testosterone and cortisol responses as well as a possible difference in these hormones between fixed and reserved players. For this purpose, two teams were purposefully selected from semi-final games of Tehran first clubs. The blood samples of 22 female players (mean age: 20 years, mean testosterone at rest: 0.507 ng/ml, mean cortisol at rest: 96.143 ng/ml) were gathered two hours before the game and 15 minutes after the game. The time each player actively participated in the game was recorded for each team separately and the collected data were analyzed by dependent and independent t tests. The results showed a significant increase in testosterone and cortisol of fixed players of both teams who actively played in 70% of the game (p<0.05). There was no relationship between the changes of testosterone and cortisol during the game and win and loss (p>0.05). In conclusion, it can be mentioned that an official basketball game with mental and physical challenges can dramatically increase testosterone hormones with strong anabolic effects as well as serum cortisol with catabolic effects in female players. Considering the physiological changes after basketball games can help to achieve suitable results and to improve players' health.