title: Down-regulation of Ras-related protein Rab 5C-dependent endocytosis and glycolysis in cisplatin-resistant ovarian cancer cell lines.
abstract: Drug resistance poses a major challenge to ovarian cancer treatment. Understanding mechanisms of drug resistance is important for finding new therapeutic targets. In the present work, a cisplatin-resistant ovarian cancer cell line A2780-DR was established with a resistance index of 6.64. The cellular accumulation of cisplatin was significantly reduced in A2780-DR cells as compared with A2780 cells consistent with the general character of drug resistance. Quantitative proteomic analysis identified 340 differentially expressed proteins between A2780 and A2780-DR cells, which involve in diverse cellular processes, including metabolic process, cellular component biogenesis, cellular processes, and stress responses. Expression levels of Ras-related proteins Rab 5C and Rab 11B in A2780-DR cells were lower than those in A2780 cells as confirmed by real-time quantitative PCR and Western blotting. The short hairpin (sh)RNA-mediated knockdown of Rab 5C in A2780 cells resulted in markedly increased resistance to cisplatin whereas overexpression of Rab 5C in A2780-DR cells increases sensitivity to cisplatin, demonstrating that Rab 5C-dependent endocytosis plays an important role in cisplatin resistance. Our results also showed that expressions of glycolytic enzymes pyruvate kinase, glucose-6-phosphate isomerase, fructose-bisphosphate aldolase, lactate dehydrogenase, and phosphoglycerate kinase 1 were down-regulated in drug resistant cells, indicating drug resistance in ovarian cancer is directly associated with a decrease in glycolysis. Furthermore, it was found that glutathione reductase were up-regulated in A2780-DR, whereas vimentin, HSP90, and Annexin A1 and A2 were down-regulated. Taken together, our results suggest that drug resistance in ovarian cancer cell line A2780 is caused by multifactorial traits, including the down-regulation of Rab 5C-dependent endocytosis of cisplatin, glycolytic enzymes, and vimentin, and up-regulation of antioxidant proteins, suggesting Rab 5C is a potential target for treatment of drug-resistant ovarian cancer. This constitutes a further step toward a comprehensive understanding of drug resistance in ovarian cancer.
authors.affiliation: §The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China;
authors.forename: Lixu
authors.initials: L
authors.lastname: Jin
authors.affiliation: From the ‡School of Life Sciences, Tsinghua University, Beijing, China;
authors.forename: Yi
authors.initials: Y
authors.lastname: Huo
authors.affiliation: ¶Zhejiang Tumor Hospital, Hangzhou, China.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Zheng
authors.affiliation: From the ‡School of Life Sciences, Tsinghua University, Beijing, China;
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Jiang
authors.affiliation: §The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China;
authors.forename: Haiyun
authors.initials: H
authors.lastname: Deng
authors.affiliation: From the ‡School of Life Sciences, Tsinghua University, Beijing, China;
authors.forename: Yuling
authors.initials: Y
authors.lastname: Chen
authors.affiliation: §The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China;
authors.forename: Qingquan
authors.initials: Q
authors.lastname: Lian
authors.affiliation: §The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, China;
authors.forename: Renshan
authors.initials: R
authors.lastname: Ge
authors.affiliation: From the ‡School of Life Sciences, Tsinghua University, Beijing, China; dht@mail.tsinghua.edu.cn.
authors.forename: Haiteng
authors.initials: H
authors.lastname: Deng
mesh_terms: D017305:Annexin A1 / Q000096:biosynthesis; D017306:Annexin A2 / Q000096:biosynthesis; D000970:Antineoplastic Agents / Q000494:pharmacology; D000975:Antioxidants / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000378:metabolism / Q000494:pharmacology*; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm; D004705:Endocytosis / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling; D005980:Glutathione Reductase / Q000096:biosynthesis; D006019:Glycolysis / Q000235:genetics*; D018841:HSP90 Heat-Shock Proteins / Q000096:biosynthesis; D006801:Humans; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000473:pathology; D040901:Proteomics; D034622:RNA Interference; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species / Q000378:metabolism; D014746:Vimentin / Q000096:biosynthesis; D020696:rab5 GTP-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: C472822:ANXA2 protein, human; D017305:Annexin A1; D017306:Annexin A2; D000970:Antineoplastic Agents; D000975:Antioxidants; D018841:HSP90 Heat-Shock Proteins; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D014746:Vimentin; D005980:Glutathione Reductase; C427755:RAB5C protein, human; D020696:rab5 GTP-Binding Proteins; D002945:Cisplatin
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 29;99(22):14298-302
references.pmid: 12370430
references.citation: Hum Cell. 2001 Dec;14(4):305-15
references.pmid: 11925933
references.citation: Mol Cell Proteomics. 2006 Mar;5(3):433-43
references.pmid: 16319398
references.citation: Oncol Res. 1995;7(12):619-24
references.pmid: 8704279
references.citation: J Clin Oncol. 2009 Jun 1;27(16):2686-91
references.pmid: 19332726
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Mar 7;832(2):286-91
references.pmid: 16483858
references.citation: Mol Cancer Res. 2008 Sep;6(9):1499-506
references.pmid: 18723829
references.citation: Nat Rev Cancer. 2007 Aug;7(8):573-84
references.pmid: 17625587
references.citation: Cancer Res. 1992 Apr 1;52(7):1895-901
references.pmid: 1551118
references.citation: PLoS One. 2010 Jun 18;5(6):e11198
references.pmid: 20585448
references.citation: Nature. 2008 Feb 28;451(7182):1116-20
references.pmid: 18264087
references.citation: Mol Cancer Ther. 2009 Apr;8(4):771-8
references.pmid: 19372549
references.citation: Cancer Res. 2005 Jan 15;65(2):613-21
references.pmid: 15695406
references.citation: J Biol Chem. 2003 Nov 14;278(46):45082-93
references.pmid: 12949081
references.citation: Proteomics. 2010 Nov;10(21):3789-99
references.pmid: 20957754
references.citation: Clin Cancer Res. 2004 Oct 1;10(19):6744-9
references.pmid: 15475465
references.citation: Int J Cancer. 2011 Oct 15;129(8):1914-22
references.pmid: 21140452
references.citation: Mol Cell Biochem. 2011 Feb;348(1-2):141-7
references.pmid: 21080034
references.citation: Nature. 2008 Mar 13;452(7184):230-3
references.pmid: 18337823
references.citation: Mol Pharmacol. 2002 Nov;62(5):1154-9
references.pmid: 12391279
references.citation: Nature. 2011 Jun 29;474(7353):609-15
references.pmid: 21720365
references.citation: CA Cancer J Clin. 2009 Jul-Aug;59(4):225-49
references.pmid: 19474385
references.citation: Cell Mol Life Sci. 2011 Sep;68(18):3033-46
references.pmid: 21637948
references.citation: PLoS One. 2012;7(7):e40717
references.pmid: 22792399
references.citation: J Cell Sci. 2007 Nov 15;120(Pt 22):3905-10
references.pmid: 17989088
references.citation: Pharm Res. 2012 Nov;29(11):3022-32
references.pmid: 22584948
references.citation: Br J Cancer. 2004 Jul 19;91(2):270-6
references.pmid: 15199393
references.citation: J Proteome Res. 2007 Feb;6(2):772-80
references.pmid: 17269733
references.citation: Oncol Rep. 2009 Jan;21(1):199-204
references.pmid: 19082462
references.citation: Biochim Biophys Acta. 2009 Feb;1794(2):225-36
references.pmid: 18973835
references.citation: J Clin Invest. 1994 Aug;94(2):703-8
references.pmid: 8040325
references.citation: Anticancer Res. 2001 Mar-Apr;21(2A):1129-33
references.pmid: 11396151
references.citation: Anticancer Agents Med Chem. 2013 Jan;13(1):159-70
references.pmid: 22931413
references.citation: Proteomics. 2005 Feb;5(3):796-804
references.pmid: 15682461
references.citation: Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3070-4
references.pmid: 1348364
references.citation: Cancer. 1987 Oct 15;60(8 Suppl):2075-80
references.pmid: 3308067
references.citation: Cancer Res. 2003 Mar 15;63(6):1311-6
references.pmid: 12649192
references.citation: J Natl Cancer Inst. 1994 Jan 5;86(1):18-24
references.pmid: 7505830
references.citation: Cancer Res. 2011 Jul 1;71(13):4412-22
references.pmid: 21571862
references.citation: Nature. 2008 Feb 28;451(7182):1111-5
references.pmid: 18264088
references.citation: Clin Cancer Res. 2006 Oct 1;12(19):5817-25
references.pmid: 17020989
references.citation: Toxicology. 2009 May 2;259(1-2):18-24
references.pmid: 19428939
references.citation: Ann N Y Acad Sci. 2012 Oct;1271:58-67
references.pmid: 23050965
references.citation: J Proteome Res. 2012 Sep 7;11(9):4605-14
references.pmid: 22900918
references.citation: Cancer Res. 2008 Apr 15;68(8):2581-6
references.pmid: 18413725
references.citation: Cancer Lett. 2007 Jan 8;245(1-2):195-204
references.pmid: 16516375
references.citation: Cancer Invest. 2011 Jan;29(1):21-8
references.pmid: 21166495
references.citation: Crit Rev Oncol Hematol. 2005 Jan;53(1):13-23
references.pmid: 15607932
references.citation: Clin Cancer Res. 2001 Oct;7(10):3193-8
references.pmid: 11595714
references.citation: Neoplasia. 2011 Nov;13(11):1069-80
references.pmid: 22131882
references.citation: Clin Cancer Res. 2008 Mar 1;14(5):1291-5
references.pmid: 18316546
references.citation: BMC Med Genomics. 2009 Jun 08;2:34
references.pmid: 19505326
references.citation: J Proteome Res. 2009 Apr;8(4):1902-12
references.pmid: 19714814
references.citation: Cancer Chemother Pharmacol. 2004 Mar;53(3):239-46
references.pmid: 14648017