Identification of up- and down-regulated proteins in doxorubicin-resistant uterine cancer cells: reticulocalbin-1 plays a key role in the development of doxorubicin-associated resistance

Eugenie Wong Soon May; Szu-Ting Lin; Chi-Chen Lin; Jo-Fan Chang; Eric Hung; Yi-Wen Lo; Li-Hsun Lin; Ren-Yu Hu; Chi-Lun Feng; Dai-Ying Lin; Shine-Bei Wu; Wen-Chi Lee; Kevin W Lyu; Hsiu-Chuan Chou; Hong-Lin Chan

doi:10.1016/j.phrs.2014.08.007

Identification of up- and down-regulated proteins in doxorubicin-resistant uterine cancer cells: reticulocalbin-1 plays a key role in the development of doxorubicin-associated resistance

Pharmacol Res. 2014 Dec:90:1-17. doi: 10.1016/j.phrs.2014.08.007. Epub 2014 Sep 19.

Authors

Affiliations

¹ Institute of Bioinformatics and Structural Biology and Department of Medical Sciences, National Tsing Hua University, Hsinchu, Taiwan.
² Institute of Biomedical Science, National Chung-Hsing University, Taichung, Taiwan; Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung-Hsing University, Taiwan; Department of Medical Research and Education, Taichung Veterans General Hospital, Taichung, Taiwan; Division of Chest Medicine, Department of Internal Medicine, Changhua Christian Hospital, Changhua, Taiwan.
³ Department of Applied Science, National Hsinchu University of Education, Hsinchu, Taiwan.
⁴ Lutheran Medical Center, Brooklyn, NY, USA; Global Scholars Program, St. George's University/Northumbria University, Newcastle upon Tyne, United Kingdom.
⁵ Department of Applied Science, National Hsinchu University of Education, Hsinchu, Taiwan. Electronic address: hlchan@life.nthu.edu.tw.
⁶ Institute of Bioinformatics and Structural Biology and Department of Medical Sciences, National Tsing Hua University, Hsinchu, Taiwan. Electronic address: chouhc@mail.nhcue.edu.tw.

PMID: 25242635
DOI: 10.1016/j.phrs.2014.08.007

Abstract

Drug resistance is a frequent cause of failure in cancer chemotherapy treatments. In this study, a pair of uterine sarcoma cancer lines, MES-SA, and doxorubicin-resistant partners, MES-SA/DxR-2μM cells and MES-SA/DxR-8μM cells, as a model system to investigate resistance-dependent proteome alterations and to identify potential therapeutic targets. We used two-dimensional differential gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) to perform this research and the results revealed that doxorubicin-resistance altered the expression of 208 proteins in which 129 identified proteins showed dose-dependent manners in response to the levels of resistance. Further studies have used RNA interference, H2A.X phosphorylation assay, cell viability analysis, and analysis of apoptosis against reticulocalbin-1 (RCN1) proteins, to prove its potency on the formation of doxorubicin resistance as well as the attenuation of doxorubicin-associated DNA double strand breakage. To sum up, our results provide useful diagnostic markers and therapeutic candidates such as RCN1 for the treatment of doxorubicin-resistant uterine cancer.

Keywords: DIGE; Doxorubicin; Resistance; Reticulocalbin-1; Uterine cancer.

Publication types

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

MeSH terms

Antibiotics, Antineoplastic / pharmacology*
Apoptosis / drug effects
Calcium-Binding Proteins / genetics
Calcium-Binding Proteins / metabolism*
Cell Line, Tumor
Down-Regulation / drug effects
Doxorubicin / pharmacology*
Drug Resistance, Neoplasm / physiology*
Electrophoresis, Gel, Two-Dimensional
Female
Humans
Proteome
RNA, Small Interfering / administration & dosage
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Up-Regulation / drug effects
Uterine Neoplasms / metabolism*

Substances

Antibiotics, Antineoplastic
Calcium-Binding Proteins
Proteome
RCN1 protein, human
RNA, Small Interfering
Doxorubicin