A mechano-reciprocal interaction plays a critical role for cancer cells searching for favorable metastasis sites. For this study, we utilized nanoscaffolds that can control the maturation of focal adhesions in order to investigate how cancer cells mechanically respond to their nanoenvironments. We found that prostate cancer cells showed linearly decreasing proliferation rate and mechanical stiffness as the size of nanoislands on nanoscaffolds where the cells were grown decreases. This mechanical signature was exacerbated for less metastatic prostate cancer cells. However, there was no dependence of mechanical responses on the geometric properties of nanoscaffolds for breast cancer cells, despite the acute inhibition of adhesion and the abrupt mechanical changes. We believe that our holistic approach that utilizes atomic force microscopy (AFM) and nanoscaffolds can reveal which mechano-reciprocal interactions are crucial for metastasis and, thus, provide useful information for anti-cancer drug development targeting integrin-associated signaling.