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Overexpression of tetraspanins affects multiple myeloma cell survival and invasive potential

Tali Tohami^*,, Liat Drucker^*,,1, Hava Shapiro, Judith Radnay and Michael Lishner^*,,

^* Oncogenetic and

Hematological Laboratories,

Department of Internal Medicine, Meir Medical Center,

Kfar Saba and Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel

¹Correspondence: Oncogenetic Laboratory, Meir Medical Center, Kfar Saba 44281, Israel. E-mail: druckerl{at}clalit.org.il

Cellular interactions with microenvironmental components are critical in multiple myeloma (MM) and impede effective disease treatment. Membranal-embedded tetraspanins, associated with metastasis suppression, are underexpressed in MM. We aimed to investigate the consequences of CD81/CD82 tetraspanins over-expression in MM cell lines. CAG and RPMI 8226 were transfected with pEGFP-N1/C1 fusion vectors of CD81/CD82. Employing flow cytometry, immunocytochemistry, and activity assays we assessed transfected cells for: morphology, survival, death, caspases, cell cycle, proliferation, oxidative stress, adhesion, motility and invasion. Overexpressed CD81/CD82 pEGFP-N1 vectors reduced survival without elevation of pre-G1 or AnnexinV+/7AAD- and independently of caspases. Decreased Ki67 and elevated intracellular glutathione were detected. No perturbations in cell cycle distribution were observed. The pEGFP-C1 vectors of CD81/CD82 caused reduction of MM cell adherence with/without fibronectin, insulin-like growth factor (IGF)-I, and matrigel. They also reduced cell motility and attenuated invasion potential, expressed by reduced secreted MMP-9 activity. These novel findings delineate the significance of CD81/CD82 expression to MM cell survival and their negative effects on cell adhesion, motility, and invasion thus, supporting their role as tumor metastasis suppressors.—Tohami T., Drucker L., Shapiro H., Radnay J., and Lishner M. Overexpression of tetraspanins affects multiple myeloma cell survival and invasive potential

Key Words: CD82 • CD81 • CAG • RPMI 8226