Addressing the role of vimentin in monocyte/macrophage differentiation using RNAi approach

MACEČKOVÁ, Vendula, Petr BENEŠ and Jan ŠMARDA. Addressing the role of vimentin in monocyte/macrophage differentiation using RNAi approach. In RNA club. České Budějovice: Přírodovědecká fakulta, Univerzita Karlova v Praze, 2005, p. 9. ISBN 1214-8598.

Basic information

• Original name: Addressing the role of vimentin in monocyte/macrophage differentiation using RNAi approach
• Name in Czech: Výzkum úlohy vimentinu v monocytové/makrofágové diferenciační dráze založený na přístupu RNAi
• Name (in English): Addressing the role of vimentin in monocyte/macrophage differentiation using RNAi approach
• Authors: MACEČKOVÁ, Vendula (203 Czech Republic), Petr BENEŠ (203 Czech Republic) and Jan ŠMARDA (203 Czech Republic, guarantor).
• Edition: České Budějovice, RNA club, p. 9-9, 2005.
• Publisher: Přírodovědecká fakulta, Univerzita Karlova v Praze

Other information

• Original language: Czech
• Type of outcome: Proceedings paper
• Field of Study: 30200 3.2 Clinical medicine
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• RIV identification code: RIV/00216224:14310/05:00012709
• Organization unit: Faculty of Science
• ISBN: 1214-8598
• Keywords in English: vimentin; differentiation; RNAi; phagocytosis
• Tags: differentiation, phagocytosis, RNAi, vimentin

Abstract

Vimentin is one of the IF proteins that undergo significant changes both in expression, structure and subcellular location during maturation of hematopoietic cells. The v-myb-transformed monoblasts of the cell line BM2 can be induced to differentiate towards macrophage-like cells using phorbol ester TPA and histon deacetylase inhibitor trichostatin A (TSA). We described earlier that level of vimentin increases during TPA- or TSA-induced differentiation of BM2 cells. In this study, RNAi technology was used to explore the role of vimentin in differentiation of BM2 cells. The cells were transfected with vector expressing siRNA targeted to endogenous vimentin mRNA and several independ clones of stable transfectants exhibiting 70% reduction of cellular vimentin level were obtained. The lack of vimentin altered morphology of the cells, reduced their phagocytic activity and production of ROS upon treatment with TPA or TSA. Our results demonstrate that up-regulation of vimentin is essential for formation of fully active macrophages during terminal steps of monocyte/macrophage differentiation.

Abstract (in English)

Vimentin is one of the IF proteins that undergo significant changes both in expression, structure and subcellular location during maturation of hematopoietic cells. The v-myb-transformed monoblasts of the cell line BM2 can be induced to differentiate towards macrophage-like cells using phorbol ester TPA and histon deacetylase inhibitor trichostatin A (TSA). We described earlier that level of vimentin increases during TPA- or TSA-induced differentiation of BM2 cells. In this study, RNAi technology was used to explore the role of vimentin in differentiation of BM2 cells. The cells were transfected with vector expressing siRNA targeted to endogenous vimentin mRNA and several independ clones of stable transfectants exhibiting 70% reduction of cellular vimentin level were obtained. The lack of vimentin altered morphology of the cells, reduced their phagocytic activity and production of ROS upon treatment with TPA or TSA. Our results demonstrate that up-regulation of vimentin is essential for formation of fully active macrophages during terminal steps of monocyte/macrophage differentiation.

Links

• GA301/03/1055, research and development project: Name: Studium molekulárních mechanismů způsobujících supresi v-Myb s využitím přístupů proteomiky

Investor: Czech Science Foundation, Study of molecular mechanisms causing v-Myb suppression using proteomics-based approach

• GP301/03/D022, research and development project: Name: Úloha vimentinu v monocytické/makrofágové diferenciační dráze

Investor: Czech Science Foundation, The role of vimentin inb the monocyte/macrophage differentiation pathway

• MSM0021622415, plan (intention): Name: Molekulární podstata buněčných a tkáňových regulací

Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations