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Preparation of Mitochondria from Ovarian Cancer Tissues and Control Ovarian Tissues for Quantitative Proteomics Analysis
Published on: November 18, 2019
Identification and validation of differentially expressed proteins in epithelial ovarian cancers using quantitative
Hong Qu1, Yuling Chen2,3, Guangming Cao1
1Department of Obstetrics & Gynecology, Beijing Chao-yang Hospital Affiliated to Capital Medical University, Beijing, China.
Abstract:
Ovarian cancer is the most lethal gynecological malignant tumor because of its high recurrence rate. In the present work, in order to find new therapeutic targets, we identified 8480 proteins in thirteen pairs of ovarian cancer tissues and normal ovary tissues through quantitative proteomics. 498 proteins were found to be differentially expressed in ovarian cancer, which involved in various cellular processes, including metabolism, response to stimulus and biosynthetic process. The expression levels of chloride intracellular channel protein 1 (CLIC1) and lectin galactoside-binding soluble 3 binding protein (LGALS3BP) in epithelial ovarian cancer tissues were significantly higher than those in normal ovary tissues as confirmed by western blotting and immunohistochemistry. The knockdown of CLIC1 in A2780 cell line downregulated expression of CTPS1, leading to the decrease of CTP and an arrest of cell cycle G1 phase, which results into a slower proliferation. CLIC1-knockdown can also slow down the tumor growth in vivo. Besides, CLIC1-knockdown cells showed an increased sensitivity to hydrogen peroxide and cisplatin, suggesting that CLIC1 was involved in regulation of redox and drug resistance in ovarian cancer cells. These results indicate CLIC1 promotes tumorgenesis, and is a potential therapeutic target in epithelial ovarian cancer treatment.
Insights
Chloride intracellular channel protein 1 (CLIC1) is overexpressed in ovarian cancer, promoting tumor growth and drug resistance. Targeting CLIC1 may offer a new therapeutic strategy for epithelial ovarian cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Proteomics
Background:
- Ovarian cancer is a lethal gynecological malignancy with high recurrence rates.
- Identifying novel therapeutic targets is crucial for improving patient outcomes.
Purpose of the Study:
- To identify novel therapeutic targets for epithelial ovarian cancer.
- To investigate the role of differentially expressed proteins in ovarian cancer progression.
Main Methods:
- Quantitative proteomics was used to analyze protein expression in 13 pairs of ovarian cancer and normal ovary tissues.
- Western blotting and immunohistochemistry confirmed the expression of key proteins.
- Gene knockdown experiments (CLIC1) were performed in vitro and in vivo.
Main Results:
- 8480 proteins were identified, with 498 found to be differentially expressed in ovarian cancer.
- Chloride intracellular channel protein 1 (CLIC1) and LGALS3BP showed significantly higher expression in ovarian cancer tissues.
- CLIC1 knockdown reduced cell proliferation, slowed tumor growth, and increased sensitivity to oxidative stress and cisplatin, indicating its role in redox regulation and drug resistance.
Conclusions:
- CLIC1 is upregulated in epithelial ovarian cancer and promotes tumorigenesis.
- CLIC1 is a potential therapeutic target for ovarian cancer treatment.
