Fluorescence molecular tomography (FMT) as a noninvasive imaging modality has been widely used for biomedical preclinical applications. However, FMT reconstruction suffers from severe ill-posedness, especially when a limited number of projections are used. In order to improve the quality of FMT reconstruction results, a discrete cosine transform (DCT) based reweighted L1-norm regularization algorithm is proposed. In each iteration of the reconstruction process, different reweighted regularization parameters are adaptively assigned according to the values of DCT coefficients to suppress the reconstruction noise. In addition, the permission region of the reconstructed fluorophores is adaptively constructed to increase the convergence speed. In order to evaluate the performance of the proposed algorithm, physical phantom and in vivo mouse experiments with a limited number of projections are carried out. For comparison, different L1-norm regularization strategies are employed. By quantifying the signal-to-noise ratio (SNR) of the reconstruction results in the phantom and in vivo mouse experiments with four projections, the proposed DCT-based reweighted L1-norm regularization shows higher SNR than other L1-norm regularizations employed in this work.