Exosome uptake through clathrin-mediated endocytosis and macropinocytosis and mediating miR-21 delivery

Tian Tian; Yan-Liang Zhu; Yue-Yuan Zhou; Gao-Feng Liang; Yuan-Yuan Wang; Fei-Hu Hu; Zhong-Dang Xiao

doi:10.1074/jbc.M114.588046

Exosome uptake through clathrin-mediated endocytosis and macropinocytosis and mediating miR-21 delivery

J Biol Chem. 2014 Aug 8;289(32):22258-67. doi: 10.1074/jbc.M114.588046. Epub 2014 Jun 20.

Authors

Tian Tian¹, Yan-Liang Zhu², Yue-Yuan Zhou², Gao-Feng Liang², Yuan-Yuan Wang², Fei-Hu Hu², Zhong-Dang Xiao³

Affiliations

¹ From the State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China and the Department of Neurobiology, Nanjing Medical University, Nanjing 210029, China.
² From the State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China and.
³ From the State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China and zdxiao@seu.edu.cn.

Abstract

Exosomes are nanoscale membrane vesicles secreted from many types of cells. Carrying functional molecules, exosomes transfer information between cells and mediate many physiological and pathological processes. In this report, utilizing selective inhibitors, molecular tools, and specific endocytosis markers, the cellular uptake of PC12 cell-derived exosomes was imaged by high-throughput microscopy and statistically analyzed. It was found that the uptake was through clathrin-mediated endocytosis and macropinocytosis. Furthermore, PC12 cell-derived exosomes can enter and deliver microRNAs (miRNAs) into bone marrow-derived mesenchymal stromal cells (BMSCs), and decrease the expression level of transforming growth factor β receptor II (TGFβRII) and tropomyosin-1 (TPM1) through miR-21. These results show the pathway of exosome internalization and demonstrate that tumor cell-derived exosomes regulate target gene expression in normal cells.

Keywords: Cell-Cell Interaction; Endocytosis; Exosome; Extracellular Vesicles; microRNA (miRNA).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biological Transport, Active
Caveolae / metabolism
Caveolin 1 / antagonists & inhibitors
Caveolin 1 / genetics
Caveolin 1 / metabolism
Cell Line
Clathrin Heavy Chains / antagonists & inhibitors
Clathrin Heavy Chains / genetics
Clathrin Heavy Chains / metabolism*
Dynamin II / antagonists & inhibitors
Dynamin II / genetics
Dynamin II / metabolism
Endocytosis
Exosomes / metabolism*
Gene Expression Regulation
Gene Knockdown Techniques
Mesenchymal Stem Cells / metabolism
MicroRNAs / genetics*
MicroRNAs / metabolism*
PC12 Cells
Phagocytosis
Pinocytosis
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism
Rats
Receptor, Transforming Growth Factor-beta Type II
Receptors, Transforming Growth Factor beta / genetics
Receptors, Transforming Growth Factor beta / metabolism
Tropomyosin / genetics
Tropomyosin / metabolism

Substances

Cav1 protein, rat
Caveolin 1
MicroRNAs
Receptors, Transforming Growth Factor beta
Tpm1 protein, rat
Tropomyosin
mirn21 microRNA, rat
Clathrin Heavy Chains
Protein Serine-Threonine Kinases
Receptor, Transforming Growth Factor-beta Type II
Dynamin II