BMVC4 is a G-quadruplex stabilizer. MVC4-treated cancer cells showed typical senescence phenotypes. BMVC4 induced senescence in both ALT and telomerase-overexpressing cells, suggesting that telomere shortening through telomerase inhibition might not be the cause for senescence. BMVC4 induced senescence by activation of pathways of response to DNA damage that was independent of its telomerase inhibitory activity. BMVC4 has the potential to be developed as a chemotherapeutic agent against both telomerase positive and ALT cancer cells.