In the absence of any treatment, spheroids belonging to the control group exhibited an increase in their area by more than 10-fold in merely 5 days of incubation in complete DMEM as clearly seen in Figure 6B. Since PTEN plasmid encapsulated in PL-NANO acts by inhibiting the in vitro tumor growth, a similar trend was observed in the 3D MCTS. This evidence concerns the gene PTEN and neoplasm.