Induction of multiple drug resistance in HMEC-1 endothelial cells after long-term exposure to sunitinib

Abstract

Limin Huang,1,* Chaoquan Hu,2,* Mélanie Di Benedetto,3 Rémi Varin,4 Jielin Liu,2,3 Li Wang,3 Jean-Pierre Vannier,4 Jian Jin,3,5 Anne Janin,3,6,7 He Lu,3,6 Hong Li4 1Department of Oncology, People’s Hospital of Guizhou Province, Guiyang, People’s Republic of China; 2Department of Surgery, Affiliated Hospital, Guiyang Medical University, Guiyang, People’s Republic of China; 3INSERM UMR-S 1165, Paris, France; 4MERCI (EA 3829), Faculté de Médecine et de Pharmacie, Université de Rouen, Rouen, France; 5School of Medicine and Pharmaceutics, Jiangnan University, Wuxi, People’s Republic of China; 6Université Paris Diderot, Laboratoire de Pathologie, Paris, France; 7AP-HP-Hôpital Saint-Louis, Laboratoire de Pathologie, Paris, France *These authors contributed equally to this studyAbstract: Multiple drug resistance is still an unsolved problem in cancer therapy. Our previous study demonstrated that the chemotherapeutic drug doxorubicin (Dox) induced upregulation of P-glycoprotein (P-gp) in endothelial cells, resulting in a 20-fold increase in drug resistance and reduced efficiency of Dox treatment in a mice tumor model. In this study, we exposed human microvascular endothelial cells (HMEC-1) to sunitinib, a tyrosine kinase receptor inhibitor, to induce drug resistance. The results show that sunitinib treatment induced multiple drug resistance in these cells. They became resistant not only to sunitinib but also to Dox, paclitaxel, and vinblastine. Significant increases in P-gp (9.3-fold), ABCG2 (breast cancer resistance protein, 1.9-fold), and multidrug resistance-associated protein 1 (2.7-fold) gene transcription were found by quantitative polymerase chain reaction quantification, and their protein expression was confirmed by Western blot. These increases gave rise to an approximately five-fold increase in half maximal inhibitory concentration in these cells in response to sunitinib treatment in vitro. The inhibitors of adenosine triphosphate-binding cassette transporters did not reverse the drug resistance in sunitinib-resistant HMEC-1 cells, assumedly because of a blockage of the pump function caused by sunitinib. Our study indicates that the antiangiogenic drug sunitinib induces multiple drug resistance in endothelial cells. The induction of adenosine triphosphate-binding cassette transporters seems not to be responsible for observed multiple drug resistance, and the underlying mechanisms remain unknown.Keywords: drug resistance, endothelial cells, cancer therapy, ABC family, sunitinib