title: Quantitative proteomic analysis of mitochondria from human ovarian cancer cells and their paclitaxel-resistant sublines.
abstract: Paclitaxel resistance is a major obstacle for the treatment of ovarian cancer. The chemoresistance mechanisms are partly related to the mitochondria. Identification of the relevant proteins in mitochondria will help in clarifying the possible mechanisms and in selecting effective chemotherapy for patients with paclitaxel resistance. In the present study, mitochondria from two paclitaxel-sensitive human ovarian cancer cell lines (SKOV3 and A2780) and their corresponding resistant cell lines (SKOV3-TR and A2780-TR) were isolated. Guanidine-modified acetyl-stable isotope labeling and liquid chromatography-hybrid linear ion trap Fourier-transform ion cyclotron resonance mass spectrometry (LC-FTICR MS) were performed to find the expressed differential proteins. Comparative proteomic analysis revealed eight differentially expressed proteins in the ovarian cancer cells and their paclitaxel-resistant sublines. Among them, mimitin and 14-3-3 ζ/δ were selected for further research. The effects of mimitin and 14-3-3 ζ/δ were explored using specific siRNA interference in ovarian cancer cell lines and immunohistochemistry in human tissue specimens. The downregulation of mimitin and 14-3-3 ζ/δ using specific siRNA in paclitaxel-resistant ovarian cancer cells led to an increase in the resistance index to paclitaxel. 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). Mimitin and 14-3-3 protein ζ/δ are potential markers of paclitaxel resistance and prognostic factors in ovarian cancer.
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.|Department of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Huang
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.|Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, China.
authors.forename: Haojie
authors.initials: H
authors.lastname: He
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing, China.
authors.forename: Wantao
authors.initials: W
authors.lastname: Ying
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing, China.|Central Laboratory, Yantai Yuhuangding Hospital, Yantai, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.|Department of Gynecological Oncology, Jiangsu Cancer Hospital and Jiangsu Institute of Cancer Research, Nanjing, China.
authors.forename: Zhiqin
authors.initials: Z
authors.lastname: Dai
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yin
authors.affiliation: Department of Cell Biology, Institute of Basic Medical Sciences, Beijing, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Mao
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing, China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Qian
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
authors.forename: Lingya
authors.initials: L
authors.lastname: Pan
mesh_terms: D048948:14-3-3 Proteins / Q000096:biosynthesis*; D000972:Antineoplastic Agents, Phytogenic / Q000627:therapeutic use; D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D018572:Disease-Free Survival; D019008:Drug Resistance, Neoplasm / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D053208:Kaplan-Meier Estimate; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000032:analysis / Q000096:biosynthesis*; D018832:Molecular Chaperones / Q000032:analysis / Q000096:biosynthesis*; D010051:Ovarian Neoplasms / Q000378:metabolism* / Q000401:mortality; D017239:Paclitaxel / Q000627:therapeutic use; D016016:Proportional Hazards Models; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D048948:14-3-3 Proteins; D000972:Antineoplastic Agents, Phytogenic; D014408:Biomarkers, Tumor; D024101:Mitochondrial Proteins; D018832:Molecular Chaperones; C501824:NDUFAF2 protein, human; D017239:Paclitaxel
keywords: Drug resistance; liquid chromatography-hybrid linear ion trap Fourier-transform ion cyclotron resonance mass spectrometry; mitochondria proteins; ovarian cancer; paclitaxel