Cell Death and Survival Alpha-CaMKII Plays a Critical Role in Determining the Aggressive Behavior of Human Osteosarcoma

Osteosarcoma is among the most frequently occurring primary bone tumors, primarily affecting adolescents and young adults. Despite improvements in osteosarcoma treatment, more specific molecular targets are needed as potential therapeutic options. One target of interest is a-Ca2þ/calmodulin-dependent protein kinase II (a-CaMKII), a ubiquitousmediator of Ca2þ-linked signaling, which has been shown to regulate tumor cell proliferation and differentiation. Here, we investigate the role of a-CaMKII in the growth and tumorigenicity of human osteosarcoma. We show that a-CaMKII is highly expressed in primary osteosarcoma tissue derived from 114 patients, and is expressed in varying levels in different human osteosarcoma (OS) cell lines [MG-63, N-methylN0-nitro-N-nitrosoguanidine (MNNG)/HOS, and 143B). To examine whether a-CaMKII regulates osteosarcoma tumorigenic properties, we genetically inhibiteda-CaMKII in two osteosarcoma cell lines using two different a-CaMKII shRNAs delivered by lentiviral vectors and overexpresseda-CaMKII by retrovirus. The genetic deletion of a-CaMKII by short hairpin RNA (shRNA) in MG-63 and 143B cells resulted in decreased proliferation (50% and 41%), migration (22% and 25%), and invasion (95% and 90%), respectively. The overexpression of a-CaMKII in HOS cells resulted in increased proliferation (240%), migration (640%), and invasion (10,000%). Furthermore, a-CaMKII deletion in MG-63 cells significantly reduced tumor burden in vivo (65%), whereas a-CaMKII overexpression resulted in tumor formation in a previously nontumor forming osteosarcoma cell line (HOS). Our results suggest that a-CaMKII plays a critical role in determining the aggressive phenotype of osteosarcoma, and its inhibition could be an attractive therapeutic target to combat this devastating adolescent disease. Mol Cancer Res; 11(4); 349–59. 2013 AACR.