Quantitative proteomic analysis of mitochondria from human ovarian cancer cells and their paclitaxel-resistant sublines.
Bottom Line: Identification of the relevant proteins in mitochondria will help in clarifying the possible mechanisms and in selecting effective chemotherapy for patients with paclitaxel resistance.Among them, mimitin and 14-3-3 ζ/δ were selected for further research.Multivariate analyses demonstrated that lower expression levels of the mimitin and 14-3-3 ζ/δ proteins were positively associated with shorter progression-free survival (PFS) and overall survival (OS) in patients with primary ovarian cancer (mimitin: PFS: P = 0.041, OS: P = 0.003; 14-3-3 ζ/δ: PFS: P = 0.031, OS: P = 0.011).
Affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.Show MeSH
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Mentions: The purity of the isolated mitochondria was determined using electron microscopy. The examination of multiple electron microscopic fields revealed that the preparations contained few, if any, nuclear fragments or other cytosolic organelles (Fig.2a). In addition, we used western blotting to validate the purity of the isolated mitochondria. The total cell protein extraction was used as a control. Western blot results showed that mitochondrial proteins were greatly enriched in the purified samples. A mitochondrial marker COX4 was determined in total cell proteins and mitochondrial preparations were isolated from SKOV3, A2780, SKOV3-TR and A2780-TR cell lines. In contrast, lamin-B, flotillin-1 and β-actin were only detected in the cellular proteins and were absent in the mitochondrial preparations (Fig.2b).
Affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.