KitL and its receptor c-Kit form a paracrine system that permits cells to respond to their microenvironment. The c-Kit kinase activating mutations can cause mastocytosis as well as Gastrointestinal Stromal Tumors and Leukemias. Although successful clinical treatments with kinase inhibitors, notably imatinib, a tumor cell population persist within the tumoral microenvironment. Does KitL/c-Kit contribute to the niche-based interactions even in the presence of kinase inhibition? By using a “niche model” we showed that bound-KitL might bind to and stabilize c-Kit in an active conformation, which is insensitive to imatinib but not to dasatinib. In this “mechanically active” state, c-Kit might recruit the PI3K which in turns induces a kinase-independent spreading response. Moreover, we showed that the constitutively active kinases W556A and D814V caused a non-spread phenotype which seems to be reverted with imatinib. A better understanding of tumor cell interactions is essential to design drugs that specifically target oncogenic proteins and the tumor microenvironment.