In this paper, the forming limit curve (FLC) of AA5754-O was predicted numerically through simulating the M-K approach with the consideration of initial groove orientation and investigated experimentally through the biaxial tensile tests. Firstly, the uniaxial tensile test and the equi-biaxial tensile test were performed to determine the mechanical properties and the anisotropic coefficients of Yld2000-2d yield criterion. Secondly, the numerical M-K approach considering the initial groove orientation and Yld2000-2d yield function was simulated to study the effects of the initial groove orientation and the imperfection coefficient on the predicted limit major strain. Finally, the shape of the cruciform specimen thinned at the center region was optimized for reducing the shear stress, and the optimized cruciform specimen was stretched under different loading paths to determine the experimental limit strains. Through comparing the predicted limit strain at uniaxial tensile stress state and the experimental one, the imperfection coefficient was confirmed, and a good agreement between the numerical result and experimental data was presented. This work showed that experimentally determining the limit strains through biaxial tensile test was achievable, and the proposed numerical M-K approach was effective for predicting the limit strains.