Mitochondrial proteomics with siRNA knockdown to reveal ACAT1 and MDH2 in the development of doxorubicin-resistant uterine cancer.

Abstract

Mitochondria are key organelles in mammary cells in responsible for a number of cellular functions including cell survival and energy metabolism. Moreover, mitochondria are one of the major targets under doxorubicin treatment. In this study, low-abundant mitochondrial proteins were enriched for proteomic analysis with the state-of-the-art two-dimensional differential gel electrophoresis (2D-DIGE) and matrix-assistant laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) strategy to compare and identify the mitochondrial protein profiling changes in response to the development of doxorubicin resistance in human uterine cancer cells. The mitochondrial proteomic results demonstrate more than fifteen hundred protein features were resolved from the equal amount pooled of three purified mitochondrial proteins and 101 differentially expressed spots were identified. In which, 39 out of these 101 identified proteins belong to mitochondrial proteins. Mitochondrial proteins such as acetyl-CoA acetyltransferase (ACAT1) and malate dehydrogenase (MDH2) have not been reported with the roles on the formation of doxorubicin resistance in our knowledge. Further studies have used RNA interference and cell viability analysis to evidence the essential roles of ACAT1 and MDH2 on their potency in the formation of doxorubicin resistance through increased cell viability and decreased cell apoptosis during doxorubicin treatment. To sum up, our current mitochondrial proteomic approaches allowed us to identify numerous proteins, including ACAT1 and MDH2, involved in various drug-resistance-forming mechanisms. Our results provide potential diagnostic markers and therapeutic candidates for the treatment of doxorubicin-resistant uterine cancer.

• Authors: Yi-Wen Lo, Szu-Ting Lin, Shing-Jyh Chang, Chia-Hao Chan, Kevin W Lyu, Jo-Fan Chang, Eugenie Wong Soon May, Dai-Ying Lin, Hsiu-Chuan Chou, Hong-Lin Chan
• Journal: Journal of cellular and molecular medicine (J Cell Mol Med) Vol. 19 Issue 4 Pg. 744-59 (Apr 2015) ISSN: 1582-4934 [Electronic] England
• PMID: 25639359 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2015 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Chemical References

• Antibiotics, Antineoplastic
• Mitochondrial Proteins
• Proteome
• Doxorubicin
• Malate Dehydrogenase
• ACAT1 protein, human
• Acetyl-CoA C-Acetyltransferase

Topics

• Acetyl-CoA C-Acetyltransferase (genetics, metabolism)
• Antibiotics, Antineoplastic (pharmacology)
• Apoptosis (drug effects)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Dose-Response Relationship, Drug
• Doxorubicin (pharmacology)
• Drug Resistance, Neoplasm
• Electrophoresis, Gel, Two-Dimensional
• Female
• Humans
• Immunoblotting
• Malate Dehydrogenase (genetics, metabolism)
• Mitochondrial Proteins (genetics, metabolism)
• Proteome (genetics, metabolism)
• Proteomics (methods)
• RNA Interference
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• Uterine Neoplasms (genetics, metabolism, pathology)