2,3,5-Triiodobenzoic acid (TIBA) is a well established inhibitor of polar transport in plant cells, a phenomenon that allows morphogenesis in plants, and due to this role in plant physiology it is considered as a pesticide. Another important feature of TIBA is that it can be also used as the basic structure for most of the clinically used contrast agents for computed tomography and other X-ray procedures, and more recently, it was used in nano-delivery systems. Even though there are several indications of the potential antitumoral activity of TIBA, its cell death-induced pathway has not been elucidated. The non-small cell lung cancer (NSCLC) cell line H460 was treated with the medium and TIBA at 100, 250 and 500 μM for 48 h. Also, cells were treated with zVAD and cyclosporine A (CSA), and cell cycle analysis and immunofluorescence were performed and loss of MMP was assessed. We show for the first time that TIBA itself is able to induce caspase-dependent apoptosis in lung cancer through an intrinsic pathway and that it is more active than carboplatin and betulinic acid against lung cancer. Thus, TIBA may constitute another therapeutic alternative to treat lung cancer and other tumors.