Differential spindle assembly checkpoint response in human lung adenocarcinoma cells

Abstract.

CI-980 is an antimicrotubule agent that binds the colchicine site on tubulin. We examined CI-980 cytotoxicity in two lung adenocarcinoma cell lines, A549 and A427. Depolymerization of microtubules following CI-980 treatment resulted in a mitotic arrest in the A549 population, but not in the A427 population. Similar responses were obtained following treatment with Taxol and nocodazole. Drug-treated A427 cells exited mitosis, generating a population dominated by multinucleated cells, while both multinucleated and apoptotic cells were present in the A549 population after extended drug treatment. CI-980-induced microtubule depolymerization was only partially reversible. However, regrowth of some microtubules in mitotic A549 cells following drug washout resulted in multinucleation of the population in the absence of apoptosis. These results show that A427 cells have a defective spindle assembly checkpoint. Levels of the MAD2 and BUB1 checkpoint proteins were similar in both A549 and A427 cells, suggesting that the checkpoint defect in the A427 cells is downstream of these proteins. In addition, induction of apoptosis in response to CI-980 correlates with the presence of a functional mitotic checkpoint and the extent of microtubule depolymerization.