PurposeThe main aim of this in vitro study was to evaluate both the uptake of [99Tc]Sestamibi into prostate cancer cells and the relationship among [99Tc]Sestamibi bioaccumulation, cancer cells proliferation and apoptosis.Procedures An in vitro study in which PC3 prostate cancer cell line was cultured with increasing doses of decayed sestamibi has been developed. Specifically, PC3 cells were incubated with three different concentrations of [99Tc]Sestamibi: 10 µg/mL, 1 µg/mL, and 0.1 µg/mL Expression of apototic markers such as caspase-3 and AIF, as well as the ultrastructure of PC3 cells, were evaluated at T0 and after 24, 48, 72, and 120 hours after [99Tc]Sestamibi incubation.ResultsData here reported for the first time showed the bioaccumulation of sestamibi in prostate cancer cells. As concern the cancer cell homeostasis, the treatment of PC3 cells with [99Tc]Sestamibi strongly influenced the cells proliferation. Indeed, a significant reduction in the mitosis was observed. Noteworthy, the accumulation of sestamibi in prostate cancer cells was associated with the appearance of morphological signs of apoptosis. The immunocytochemical analysis of apoptotic biomarkers, AIF and caspase 3, in prostate cancer cells treated with 10 µg/mL of [99Tc]Sestamibi for confirmed that this radiopharmaceutical can induce the canonical apoptosis.DiscussionTo the best of our knowledge, this study for the first time reported in vitro data about the uptake of sestamibi in prostate cancer cells. The evidence about the accumulation of sestamibi in prostate cancer cells and its role in the apoptosis process can open new clinical perspectives on the use of this radiopharmaceutical in both the diagnosis and treatment of prostate cancers.