Dual-energy spectral computed tomography (DESCT) is based on fast switching between high and low voltages from view to view to obtain dual-energy imaging data, and it can generate monochromatic image sets, iodine-based material decomposition images and spectral CT curves. Quantitative spectral CT parameters may be valuable for reflecting Ki-67 expression and EGFR mutation status in non-small-cell lung cancer (NSCLC). We investigated the associations among the quantitative parameters generated in DESCT and Ki-67 expression and EGFR mutation in NSCLC. We studied sixty-five NSCLC patients with preoperative DESCT scans, and their specimens underwent Ki-67 and EGFR evaluations. Statistical analyses were performed to identify the spectral CT parameters for the diagnosis of Ki-67 expression and EGFR mutation status. We found that tumour grade and the slope of the spectral CT curve in the venous phase were the independent factors influencing the Ki-67 expression level, and the area under the curve (AUC) of the slope of the spectral CT curve in the venous phase in the receiver operating characteristic analysis for distinguishing different Ki-67 expression levels was 0.901. Smoking status and the normalized iodine concentration in the venous phase were independent factors influencing EGFR mutation, and the AUC of the two-factor combination for predicting the presence of EGFR mutation was 0.807. These results show that spectral CT parameters may be useful for predicting Ki-67 expression and the presence of EGFR mutation in NSCLC.