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Overexpression of tetraspanins affects multiple myeloma cell survival and invasive potential
Tali Tohami1, Liat Drucker, Hava Shapiro
1Oncogenetic Laboratory, Meir Medical Center, Kfar Saba 44281, Israel.
Abstract:
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.
Insights
Overexpressing CD81/CD82 tetraspanins in multiple myeloma (MM) cells reduced survival and inhibited metastasis. These findings highlight tetraspanins as potential therapeutic targets for MM treatment.
Area of Science:
- Oncology
- Cell Biology
- Molecular Biology
Background:
- Cellular interactions with the microenvironment are crucial in multiple myeloma (MM), impacting treatment efficacy.
- Tetraspanins, including CD81 and CD82, are membrane proteins linked to metastasis suppression but are underexpressed in MM.
Purpose of the Study:
- To investigate the functional consequences of overexpressing CD81 and CD82 tetraspanins in MM cell lines.
- To determine the impact of CD81/CD82 overexpression on MM cell survival, proliferation, adhesion, motility, and invasion.
Main Methods:
- MM cell lines (CAG and RPMI 8226) were transfected with pEGFP-N1/C1 fusion vectors encoding CD81/CD82.
- Flow cytometry, immunocytochemistry, and activity assays were used to assess cell morphology, survival, death, caspase activity, cell cycle, proliferation, oxidative stress, adhesion, motility, and invasion.
Main Results:
- Overexpression of CD81/CD82 via pEGFP-N1 vectors decreased MM cell survival independently of caspases, reduced proliferation (Ki67), and increased intracellular glutathione.
- Transfection with pEGFP-C1 vectors for CD81/CD82 reduced MM cell adhesion to various substrates, decreased motility, and attenuated invasion potential, evidenced by reduced MMP-9 activity.
Conclusions:
- CD81/CD82 tetraspanins play a significant role in regulating multiple myeloma cell survival.
- Overexpression of CD81/CD82 negatively impacts MM cell adhesion, motility, and invasion, supporting their function as tumor metastasis suppressors.
