PIM1 and cancer: Studies have shown that MTX can interact with protein kinases, Xiaobo Wan and co-workers in 2013, conducted a study where they computationally coupled MTX with 143 kinases in active conformation, the best 5 results were tested in vitro, and PIM1 kinase validated MTX to be used as an inhibitor in cancer cells because MTX showed to possess low nanomolar inhibitory activity against PIM1 kinase and to inhibit the PIM1-mediated phosphorylation in cancer cells [41].