DMH4, A VEGFR2 Inhibitor Effectively Suppresses Proliferation, Migration And Apoptosis Of Lung Cancer Cells

Background: Lung cancer accounts for more deaths than any other cancer. Approximate 80% of all lung cancers are NSCLC(non-small-cell lung cancer). Therapy research targeting vascular endothelial growth factor(VEGF) pathway gains its popularity in recent years. DMH4(6-[4-[2-(4-Morpholinyl)ethoxy]phenyl]-3-phenylpyrazolo[1,5-a]pyrimidine) is a potent VEGF inhibitor. Compared to AMPK, it is a more effective for VEGFR and angiogenesis inhibition.Method and result: MTT assay was taken for viability of cells treated by DMH4. After 24 hours treatment of 1 ug/ml DMH4, the viability of H460, A549, MRC5 and ADSC are 75.10%、51.47%、106.74% and 109.73%, suggesting that DMH4 had more antiproliferative effect on lung cancer cell line H460 and A549, and less on fibroblast cell line MRC5 and adipose derived stem cell line ADSC. After 24 hours of DMH4 treatment of different concentration of 0.25ug/ml、0.5ug/ml、1ug/ml、2ug/ml、4ug/ml、8ug/ml and 16ug/ml, the cell viability of A549 were 77.25%、55.94%、51.47%、43.23%、42.82%、42.61%and 40.04%, indicating that the impact of DMH4 was concentration dependent. Through the scratch test, inhibition of cell migration by DMH4 treatment on H460 and A549 was shown. Cells undergoing apoptosis or necrosis were identified by a Annexin V and PI double staining kit through flowcytometry. DMH4 improved cell apoptosis of 1 ug/ml DMH4 treated H460 and A549. After treated by DMH4, phospho-VEGF receptor 2 level in H460 and A549 were down-regulated, proving DMH4 inhibited VEGFR2. Human apoptosis antibody array membrane was used to test the apoptosis pathway involved by DMH4. 43 targets were tested by this experiment, and most of the proteins involved in apoptosis pathway were shown up-regulated, suggesting DMH4 improved cell apoptosis by inhibiting VEGF pathway.Conclusion: DMH4 as a VEGFR2 inhibitor was able to inhibit lung cancer cell’s proliferation and migration and worth studying deeper.