Purpose. To evaluate signal intensity (SI) differences between 3.0 T and 1.5 T on T2-weighted (T2w), diffusion-weighted imaging (DWI), and apparent diffusion coefficient (ADC) in rectal cancer pre-, during, and postneoadjuvant chemoradiotherapy (CRT). Materials and Methods. 22 patients with locally advanced rectal cancer were prospectively enrolled. All patients underwent T2w, DWI, and ADC pre-, during, and post-CRT on both 3.0 T MRI and 1.5 T MRI. A radiologist drew regions of interest (ROIs) of the tumor and obturator internus muscle on the selected slice to evaluate SI and relative SI (rSI). Additionally, a subanalysis evaluating the SI before and after-CRT (∆SI pre-post) in complete responder patients (CR) and nonresponder patients (NR) on T2w, DWI, and ADC was performed. Results. Significant differences were observed for T2w and DWI on 3.0 T MRI compared to 1.5 T MRI pre-, during, and post-CRT (all
P
<
0.001
), whereas no significant differences were reported for ADC among all controls (all
P
>
0.05
). rSI showed no significant differences in all the examinations for all sequences (all
P
>
0.05
). ∆SI showed significant differences between 3.0 T and 1.5 T MRI for DWI-∆SI in CR and NR (
188.39
±
166.90
vs.
30.45
±
21.73
and
169.70
±
121.87
vs.
22.00
±
31.29
, respectively, all
P
0.02) and ADC-∆SI for CR (
−
0.58
±
0.27
vs.
−
0.21
±
0.24
P
value 0.02), while no significant differences were observed for ADC-∆SI in NR and both CR and NR for T2w-∆SI. Conclusion. T2w-SI and DWI-SI showed significant differences for 3.0 T compared to 1.5 T in all three controls, while ADCSI showed no significant differences in all three controls on both field strengths. rSI was comparable for 3.0 T and 1.5 T MRI in rectal cancer patients; therefore, rectal cancer patients can be assessed both at 3.0 T MRI and 1.5 T MRI. However, a significant DWI-∆SI and ADC-∆SI on 3.0 T in CR might be interpreted as a better visual assessment in discriminating response to therapy compared to 1.5 T. Further investigations should be performed to confirm future possible clinical application.