Pulsed electromagnetic field (PEMF) therapy involves the use of magnetic waveform energy for targeted treatment delivery. This technique has shown promising results in the treatment of various cancers. Currently, treatment of bladder cancer is highly invasive, involving intravesical chemotherapy or radical cystectomy. The potential therapeutic effects of PEMF therapy on bladder cancer are a relatively new and understudied area; therefore, the goal of this investigation was to gain mechanistic insight by examining the effects of PEMF therapy on a bladder cancer cell line in vitro. Cells from the bladder cancer cell line HT-1197 were cultured and incubated with (treatment group) or without (control group) PEMF therapy for one hour each day for five days. Cell counts were compared using Incucyte data to determine proliferation rates. At days 1 and 5, total RNA was isolated from cells, and following quantity and quality checks, gene expression was compared between the two groups. Proliferation rates from cell line HT-1197 were compared to prior published results on the bladder cancer cell line HT-1376. HT-1197 cells treated with PEMF therapy had slower proliferation rates compared to controls ( < 0.05), but HT-1376 cells did not ( > 0.05). Principal component analysis showed complete separation of treated and untreated cells, with PEMF treatment accounting for 76% of the variation between the groups. Expression of numerous genes and cancer-related pathways was altered in the treated cells relative to the controls. Bladder cancer HT-1197 cells treated with PEMF therapy had slower proliferation and corresponding changes in gene expression. Several cancer-relevant pathways were differentially regulated following PEMF treatment. The conclusions are limited by the lack of a control healthy urothelial cell line in the experiments. Despite this shortcoming, our results suggest that PEMF therapy may be a promising avenue for further research in the treatment of bladder cancer.