A novel game-theoretic framework for landmark-based image segmentation is presented. Landmark detection is formulated as a game, in which landmarks are players, landmark candidate points are strategies, and likelihoods that candidate points represent landmarks are payoffs, determined according to the similarity of image intensities and spatial relationships between the candidate points in the target image and their corresponding landmarks in images from the training set. The solution of the formulated game-theoretic problem is the equilibrium of candidate points that represent landmarks in the target image and is obtained by a novel iterative scheme that solves the segmentation problem in polynomial time. The object boundaries are finally extracted by applying dynamic programming to the optimal path searching problem between the obtained adjacent landmarks. The performance of the proposed framework was evaluated for segmentation of lung fields from chest radiographs and heart ventricles from cardiac magnetic resonance cross sections. The comparison to other landmark-based segmentation techniques shows that the results obtained by the proposed game-theoretic framework are highly accurate and precise in terms of mean boundary distance and area overlap. Moreover, the framework overcomes several shortcomings of the existing techniques, such as sensitivity to initialization and convergence to local optima.