Targeting MAGE-C1/CT7 expression increases cell sensitivity to the proteasome inhibitor bortezomib in multiple myeloma cell lines.
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Targeting MAGE-C1/CT7 expression increases cell sensitivity to the proteasome inhibitor bortezomib in multiple myeloma cell lines. / de Carvalho, Fabricio; Costa, Erico T; Camargo, Anamaria A; Gregorio, Juliana C; Masotti, Cibele; Andrade, Valeria C C; Strauss, Bryan E; Caballero, Otavia L; Atanackovic, Djordje; Colleoni, Gisele W B.
in: PLOS ONE, Jahrgang 6, Nr. 11, 11, 2011, S. 27707.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Targeting MAGE-C1/CT7 expression increases cell sensitivity to the proteasome inhibitor bortezomib in multiple myeloma cell lines.
AU - de Carvalho, Fabricio
AU - Costa, Erico T
AU - Camargo, Anamaria A
AU - Gregorio, Juliana C
AU - Masotti, Cibele
AU - Andrade, Valeria C C
AU - Strauss, Bryan E
AU - Caballero, Otavia L
AU - Atanackovic, Djordje
AU - Colleoni, Gisele W B
PY - 2011
Y1 - 2011
N2 - The MAGE-C1/CT7 encodes a cancer/testis antigen (CTA), is located on the chromosomal region Xq26-27 and is highly polymorphic in humans. MAGE-C1/CT7 is frequently expressed in multiple myeloma (MM) that may be a potential target for immunotherapy in this still incurable disease. MAGEC1/CT7 expression is restricted to malignant plasma cells and it has been suggested that MAGE-C1/CT7 might play a pathogenic role in MM; however, the exact function this protein in the pathophysiology of MM is not yet understood. Our objectives were (1) to clarify the role of MAGE-C1/CT7 in the control of cellular proliferation and cell cycle in myeloma and (2) to evaluate the impact of silencing MAGE-C1/CT7 on myeloma cells treated with bortezomib. Myeloma cell line SKO-007 was transduced for stable expression of shRNA-MAGE-C1/CT7. Downregulation of MAGE-C1/CT7 was confirmed by real time quantitative PCR and western blot. Functional assays included cell proliferation, cell invasion, cell cycle analysis and apoptosis. Western blot showed a 70-80% decrease in MAGE-C1/CT7 protein expression in inhibited cells (shRNA-MAGE-C1/CT7) when compared with controls. Functional assays did not indicate a difference in cell proliferation and DNA synthesis when inhibited cells were compared with controls. However, we found a decreased percentage of cells in the G2/M phase of the cell cycle among inhibited cells, but not in the controls (p<0.05). When myeloma cells were treated with bortezomib, we observed a 48% reduction of cells in the G2/M phase among inhibited cells while controls showed 13% (empty vector) and 9% (ineffective shRNA) reduction, respectively (p<0.01). Furthermore, inhibited cells treated with bortezomib showed an increased percentage of apoptotic cells (Annexin V+/PI-) in comparison with bortezomib-treated controls (p<0.001). We found that MAGE-C1/CT7 protects SKO-007 cells against bortezomib-induced apoptosis. Therefore, we could speculate that MAGE-C1/CT7 gene therapy could be a strategy for future therapies in MM, in particular in combination with proteasome inhibitors.
AB - The MAGE-C1/CT7 encodes a cancer/testis antigen (CTA), is located on the chromosomal region Xq26-27 and is highly polymorphic in humans. MAGE-C1/CT7 is frequently expressed in multiple myeloma (MM) that may be a potential target for immunotherapy in this still incurable disease. MAGEC1/CT7 expression is restricted to malignant plasma cells and it has been suggested that MAGE-C1/CT7 might play a pathogenic role in MM; however, the exact function this protein in the pathophysiology of MM is not yet understood. Our objectives were (1) to clarify the role of MAGE-C1/CT7 in the control of cellular proliferation and cell cycle in myeloma and (2) to evaluate the impact of silencing MAGE-C1/CT7 on myeloma cells treated with bortezomib. Myeloma cell line SKO-007 was transduced for stable expression of shRNA-MAGE-C1/CT7. Downregulation of MAGE-C1/CT7 was confirmed by real time quantitative PCR and western blot. Functional assays included cell proliferation, cell invasion, cell cycle analysis and apoptosis. Western blot showed a 70-80% decrease in MAGE-C1/CT7 protein expression in inhibited cells (shRNA-MAGE-C1/CT7) when compared with controls. Functional assays did not indicate a difference in cell proliferation and DNA synthesis when inhibited cells were compared with controls. However, we found a decreased percentage of cells in the G2/M phase of the cell cycle among inhibited cells, but not in the controls (p<0.05). When myeloma cells were treated with bortezomib, we observed a 48% reduction of cells in the G2/M phase among inhibited cells while controls showed 13% (empty vector) and 9% (ineffective shRNA) reduction, respectively (p<0.01). Furthermore, inhibited cells treated with bortezomib showed an increased percentage of apoptotic cells (Annexin V+/PI-) in comparison with bortezomib-treated controls (p<0.001). We found that MAGE-C1/CT7 protects SKO-007 cells against bortezomib-induced apoptosis. Therefore, we could speculate that MAGE-C1/CT7 gene therapy could be a strategy for future therapies in MM, in particular in combination with proteasome inhibitors.
KW - Humans
KW - Cell Line, Tumor
KW - Neoplasm Invasiveness
KW - Gene Silencing
KW - Cell Proliferation/drug effects
KW - Antineoplastic Agents/pharmacology
KW - Antigens, Neoplasm/genetics
KW - Apoptosis/drug effects/genetics
KW - Boronic Acids/pharmacology
KW - Cell Division/drug effects/genetics
KW - G2 Phase/drug effects/genetics
KW - Gene Expression Regulation, Neoplastic/drug effects/genetics
KW - Multiple Myeloma/genetics/pathology
KW - Neoplasm Proteins/deficiency/genetics
KW - Protease Inhibitors/pharmacology
KW - Proteasome Endopeptidase Complex/antagonists & inhibitors
KW - Pyrazines/pharmacology
KW - RNA, Small Interfering/genetics
KW - Humans
KW - Cell Line, Tumor
KW - Neoplasm Invasiveness
KW - Gene Silencing
KW - Cell Proliferation/drug effects
KW - Antineoplastic Agents/pharmacology
KW - Antigens, Neoplasm/genetics
KW - Apoptosis/drug effects/genetics
KW - Boronic Acids/pharmacology
KW - Cell Division/drug effects/genetics
KW - G2 Phase/drug effects/genetics
KW - Gene Expression Regulation, Neoplastic/drug effects/genetics
KW - Multiple Myeloma/genetics/pathology
KW - Neoplasm Proteins/deficiency/genetics
KW - Protease Inhibitors/pharmacology
KW - Proteasome Endopeptidase Complex/antagonists & inhibitors
KW - Pyrazines/pharmacology
KW - RNA, Small Interfering/genetics
U2 - 10.1371/journal.pone.0027707
DO - 10.1371/journal.pone.0027707
M3 - SCORING: Journal article
VL - 6
SP - 27707
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 11
M1 - 11
ER -
