The authors prospectively investigated the uptake and kinetics of Tc-99m tetrofosmin (Tetro) in benign and malignant lung lesions and the effect of radiotherapy, chemotherapy, or both on Tc-99m Tetro uptake in malignant lung tumors. Dynamic and planar Tetro imaging were performed in 45 patients with pulmonary lesions during a period of 28 months (34 untreated malignant tumors, 11 benign lesions). Tetro uptake was visibly increased in 26 of 34 malignant tumors, with a mean lesion to contralateral normal tissue ratio of 1.44 ± 0.29 and a tumor washout rate of 28.4 ± 6.6% 30 minutes after injection. In 3 of 11 benign lesions, Tetro uptake was observed. Of the 26 patients with malignant tumors and positive Tetro uptake, nine had repeated imaging 6 to 8 weeks after therapy. The patients were treated with radiotherapy, chemotherapy, or both. Reduction in radiological tumor size was used as the clinical response parameter. In five of nine patients, the course of Tetro uptake in follow-up imaging was in accordance with that of radiological tumor size. Two of four remaining patients had only slight discordance between the alteration of Tetro uptake and radiological tumor size. The sensitivity of Tetro to detect malignant lung lesions in our patients was 77%. The specificity and accuracy of the method were 73% and 76%, respectively. Tetro has limited diagnostic value in detecting lung cancer. It may be useful to monitor the response to therapy in malignant lung tumors with initial tracer uptake. Broader trials on this matter are needed for further clarification.