Open AccessMitochondrial proteomics with si RNA knockdown to reveal ACAT 1 and MDH 2 in the development of doxorubicin‐resistant uterine cancer
Author(s) -
Lo YiWen,
Lin SzuTing,
Chang ShingJyh,
Chan ChiaHao,
Lyu Kevin W.,
Chang JoFan,
May Eugenie Wong Soon,
Lin DaiYing,
Chou HsiuChuan,
Chan HongLin
Publication year - 2015
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/jcmm.12388
Subject(s) - mitochondrion , biology , doxorubicin , malate dehydrogenase , proteomics , viability assay , cancer cell , microbiology and biotechnology , gene knockdown , cell , biochemistry , apoptosis , cancer , enzyme , genetics , chemotherapy , gene
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 ( ACAT 1) and malate dehydrogenase ( MDH 2) 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 ACAT 1 and MDH 2 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 ACAT 1 and MDH 2, involved in various drug‐resistance‐forming mechanisms. Our results provide potential diagnostic markers and therapeutic candidates for the treatment of doxorubicin‐resistant uterine cancer.