Background Apparent diffusion coefficient (ADC) values achieve promising results in treatment response prediction in patients with several types of cancers. Purpose To determine whether ADC values predict neoadjuvant chemoradiation treatment (nCRT) response in patients with locally advanced rectal cancer (LARC). Material and Methods Forty-four patients with LARC who underwent magnetic resonance imaging scans before and after nCRT followed by delayed surgery were enrolled retrospectively. The sample was distributed as follows: responders (R), n = 8; and non-responders (Non-R), n = 36. Three markers of treatment response were considered: post-nCRT measures; ΔADC; and Δ%ADC. Statistical analysis included a Wilcoxon test, a Mann–Whitney U test, and a receiver operating characteristic (ROC) analysis in order to evaluate the diagnostic accuracy for each ADC value marker to differentiate between R and Non-R. Results Both minimum and mean ADC values were significantly higher after nCRT in the R group, while non-significant differences between basal and control ADC values were found in the non-R group. In addition, ΔADC and Δ%ADC exhibited increased values after nCRT in R when compared with non-R. ROC analysis revealed the following diagnostic performance parameters: post-nCRT: ADCmin = 1.05 × 10−3 mm2/s (sensitivity 61.1% and specificity 66.7%), ADCmean = 1.50 × 10−3 mm2/s (sensitivity 72.2% and specificity 83.3%), ΔADC: ADCmin = 0.35 (sensitivity 66.7% and specificity 83.3%), ADCmean = 0.50 (sensitivity 72% and specificity 83%); and Δ%ADC: ADCmin = 44% (sensitivity 66.7% and specificity 83.3%) and ADCmean = 60% (sensitivity 83% and specificity 99%). Conclusion Our findings suggest that post-treatment rectal tumor ADC values, as well changes between pre- and post-treatment values, may be biomarkers for predicting treatment response in patients with LARC who underwent nCRT.