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Updated: Jun 7, 2026
Preparation of Mitochondria from Ovarian Cancer Tissues and Control Ovarian Tissues for Quantitative Proteomics Analysis
Published on: November 18, 2019
Mitochondrial comparative proteomics of human ovarian cancer cells and their platinum-resistant sublines
Zhiqin Dai1, Jie Yin, Haojie He
1Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, PR China.
Abstract:
Resistance to platinum-based chemotherapy is the major obstacle to successful treatment of ovarian cancer. It is evident that mitochondrial defects and the dysfunctions of oxidative phosphorylation and energy production in ovarian cancer cells were directly related to their resistance to platinum drugs. Using 2-D DIGE, we compared mitochondrial proteins from two platinum-sensitive human ovarian cancer cell lines (SKOV3 and A2780) with that of four platinum-resistant sublines (SKOV3/CDDP, SKOV3/CBP, A2780/CDDP, and A2780/CBP). Among the 236 differentially expressed spots, five mitochondrial proteins (ATP-α, PRDX3, PHB, ETF, and ALDH) that participate in the electron transport respiratory chain were identified through mass spectrometry. All of them are downregulated in one or two of the platinum-resistant cell lines. Three proteins (ATP-α, PRDX3, and PHB) were validated by using western blot and immunohistochemistry. There is a significant decrease of PHB in tumor tissues from ovarian cancer patients who were resistant to platinum-based chemotherapies. This is the first direct mitochondrial proteomic comparison between platinum-sensitive and resistant ovarian cancer cells. These studies demonstrated that 2-D DIGE-based proteomic analysis could be a powerful tool to investigate limited mitochondrial proteins, and the association of PHB expression with platinum resistance indicates that mitochondria defects may contribute to platinum resistance in ovarian cancer cells.
Insights
Mitochondrial defects are linked to platinum resistance in ovarian cancer. Key proteins like PHB are downregulated in resistant cells and patient tumors, suggesting a role for mitochondrial dysfunction in treatment failure.
Area of Science:
- Biochemistry
- Oncology
- Proteomics
Background:
- Platinum-based chemotherapy is a primary treatment for ovarian cancer.
- Resistance to platinum drugs is a significant clinical challenge.
- Mitochondrial dysfunction, including impaired oxidative phosphorylation, is associated with platinum resistance.
Purpose of the Study:
- To compare mitochondrial proteomes between platinum-sensitive and resistant ovarian cancer cell lines.
- To identify specific mitochondrial proteins involved in platinum resistance.
- To validate the expression of identified proteins in patient tumor tissues.
Main Methods:
- Two-dimensional difference gel electrophoresis (2-D DIGE) to compare mitochondrial proteins.
- Mass spectrometry for protein identification.
- Western blot and immunohistochemistry for protein validation.
Main Results:
- Five mitochondrial proteins (ATP-α, PRDX3, PHB, ETF, ALDH) in the electron transport chain were downregulated in resistant cell lines.
- PHB expression was significantly decreased in tumor tissues from platinum-resistant ovarian cancer patients.
- This study represents the first direct mitochondrial proteomic comparison between sensitive and resistant ovarian cancer cells.
Conclusions:
- Mitochondrial proteomic analysis using 2-D DIGE is effective for identifying proteins related to drug resistance.
- Downregulation of PHB is associated with platinum resistance in ovarian cancer.
- Mitochondrial defects likely contribute to platinum resistance in ovarian cancer.
