It is still a challenge that organotin(IV) carboxylate complexes with high-performance two-photon activity for cancer therapy. At present work, two novel organotin carboxylate complexes LSn1 and LSn2, containing coumarin moiety, were rationally designed for two-photon fluorescent imaging and anticancer purpose. The complexes possessed large two-photon action cross-sections and high quantum yields. Living cells evaluation revealed that complexes LSn1 and LSn2 exhibited good biocompatibility and deep tissue penetration over femtosecond laser with wavelength of 840 nm. Furthermore, the antitumor active and as well as possible mechanism of complexes LSn1 and LSn2 have been investigated systematically. The results indicated that complexes LSn1 and LSn2 could induce apoptotic cell death through a mitochondria] dysfunction and ROS elevation pathway. The present work provides a strategy for rationally designing organotin(IV) carboxylate complexes with two-photon activity and antitumor activity.