Cancer is one of the deadliest diseases around today, so there is a need to develop new treatments that are more effective and have fewer side effects. The use of suicide gene therapy may be an attractive strategy. As we have seen previously in  other publications of ldrB gene, ldrB gene transfection of colorectal, breast and cervical cancer cells induces a clear decrease in proliferation rate and death in all cell lines. To elucidate the mechanism of action of the ldrB gene, morphological studies using scanning electron microscopy as well as cell cycle and proteomic analysis were carried out. Our results showed that ldrB gene expression induced cell cycle arrest in G0/G1 phase for both HCT-116 and MCF-7 while we observed S phase arrest for the HeLa cell line. Scanning electron microscopy showed that LdrB toxin generated a decrease in cell density and the appearance of large pores in the membrane that can lead to cell death, as well as the appearance of several bubbles and membrane rupture. Finally, western blot assay was performed, showing modification in the expression pattern of proteins involved in the apoptosis and pyroptosis pathways in the transfected cells with respect to the control. Taken together, our results provide evidence that the cytotoxic effect of the ldrB gene is mediated by different cell death pathways supporting its potential as an anticancer tool for suicide gene therapy.