Identification of Cisplatin-Resistance Associated Genes through Proteomic Analysis of Human Ovarian Cancer Cells and a Cisplatin-resistant Subline

  • Zhou, Jing (Department of Gynecology Oncology, Zhongnan Hospital of Wuhan University) ;
  • Wei, Yue-Hua (Department of Medical Imaging Central, Zhongnan Hospital of Wuhan University) ;
  • Liao, Mei-Yan (Department of Medical Imaging Central, Zhongnan Hospital of Wuhan University) ;
  • Xiong, Yan (Department of Gynecology Oncology, Zhongnan Hospital of Wuhan University) ;
  • Li, Jie-Lan (Department of Medical Imaging Central, Zhongnan Hospital of Wuhan University) ;
  • Cai, Hong-Bing (Department of Gynecology Oncology, Zhongnan Hospital of Wuhan University)
  • Published : 2012.12.31


Chemoresistance to cancer therapy is a major obstacle to the effective treatment of human cancers with cisplatin (DDP), but the mechanisms of cisplatin-resistance are not clear. In this study, we established a cisplatin-resistant human ovarian cancer cell line (COC1/DDP) and identified differentially expressed proteins related to cisplatin resistance. The proteomic expression profiles in COC1 before and after DDP treatment were examined using 2-dimensional electrophoresis technology. Differentially expressed proteins were identified using matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and high performance liquid chromatography-electrospray tandem MS (NanoUPLC-ESI-MS/MS). 5 protein spots, for cytokeratin 9, keratin 1, deoxyuridine triphosphatase (dUTPase), aarF domain containing kinase 4 (ADCK 4) and cofilin1, were identified to be significantly changed in COC1/DDP compared with its parental cells. The expression of these five proteins was further validated by quantitative PCR and Western blotting, confirming the results of proteomic analysis. Further research on these proteins may help to identify novel resistant biomarkers or reveal the mechanism of cisplatin-resistance in human ovarian cancers.


  1. Aebi S, Kurdi-Haidar B, Gordon R, et al (1996). Loss of DNA mismatch repair in acquired resistance to cisplatin. Cancer Res, 56, 3087-90.
  2. Andrea U, Julia P and Sergio B (2005). A proteomic investigation into etoposide chemo-resistance of neuroblastoma cell lines. Proteomics, 5, 796-804.
  3. Canman CE, Radany EH, Parsels LA, et al (1994). Davis MA, Lawrence TS and Maybaum J: Induction of resistance to fluorodeoxyuridine cytotoxicity and DNA damage in human tumor cells by expression of Escherichi a coli deoxyuridinetriphosphatase. Cancer Res, 54, 2296-8.
  4. Cecconi D, Donadelli M, Scarpa A, et al (2005). Proteomic analysis pancreatic ductal carcinoma cells after combined treatment with gemcitabine and trichostatin A. J Proteome Res, 4, 1909-16.
  5. Chua BT, Volbracht C, Tan KO, et al (2003). Mitochondrial translocation of cofilin is an early step in apoptosis induction. Nat Cell Biol, 5, 1083-9.
  6. DesMarais V, Ghosh M, Eddy R, et al (2005). Cofilin takes the lead. J Cell Sci, 118, 19-26.
  7. Hamada S, Kamada M, Furumoto H, et al (1994). Expression of glutathione S-transferase-pi in human ovarian cancer as an indicator of resistance to chemotherapy. Gynecol Oncol, 52, 313-9.
  8. Jemal A, Siegel R, Ward E et al (2009). Cancer statistics. CA Cancer J Clin, 59, 225-49.
  9. Kartalou M and Essigmann JM (2001). Mechanisms of resistance to cisplatin. Mutat Res, 478, 23-43.
  10. Kim SW, Kim S, Nam EJ, et al (2011). Comparative Proteomic Analysis of Advanced Serous Epithelial Ovarian Carcinoma: Possible Predictors of Chemoresistant disease. OMICS, 15, 281-92.
  11. Liu CC, Lu TC, Li HH, et al (2010). Phosphoproteomic identifi-cation and phylogenetic analysis of ribosomal P-proteins in Populus dormant terminal buds. Planta, 231, 571-81.
  12. Livak KJ and Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25, 402-8.
  13. Marth C, Widschwendter M, KaernJ, et al (1997). Cisplatin resistance is associated with reduced interfer on-gammasensitivity and increased HER-2 expression in cultured ovarian carcinoma cells. Cance, 76, 1328-32.
  14. Ozols RF and Young RC (1985). High-dose cisplatin therapy in ovarian cancer. Semin Oncol, 12, 21-30.
  15. Rudin CM, Yang Z, Schumaker LM, et al (2003). Inhibition of glutathione synthesis reverses Bcl-2-mediated cisplatin resistance. Cancer Res, 63, 312-8.
  16. Sasaki H, Sheng Y, Kotsuji F and Tsang BK (2000). Downregulation of X-linked inhibitor of apoptosis protein induces apoptosis in chemoresistant human ovarian cancer cells. Cancer Res, 60, 5659-66.
  17. Veneroni S, Zaffaroni N, Daidone MG, et al (1994). Expression of P-glycoprote-in and in vitro or in vivo resistance to doxorubicin and cisplatin in breast and ovarian cancers. Eur J Cancer, 30A, 1002-7.
  18. Wang WG, Goswami S, Lapidus K, et al (2004). Identification and testing of a gene expression signature of invasive carcinoma cells within primary mammary tumors. Cancer Res, 64: 8585-94.
  19. Wang W, Mouneimne G, Sidani M, et al (2006). The activity status of cofilin is directly related to invasion, intravasation, and metastasis of mammary tumors. J Cell Biol, 173, 395-404.
  20. Williams J, Lucas PC, Griffith KA, et al (2005). Expression of BclxL in ovarian carcinoma is associated with chemoresistance and recurrent disease. Gynecol Oncol, 96, 287-95.
  21. Wong CS, Wong VW, Chan CM, et al (2008). Identification of 5-fluorouracil response proteins in colorectal cacinoma cell line SW480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Oncol Pep, 20, 89-98.
  22. Yuan ZQ, Feldman RI, Sussman GE, et al (2003). AKT2 inhibition of cisplatin-induced JNK/p38 and Bax activation by phosphorylation of ASK1-Implication of AKT2 in chemoresistance. J Biol Chem, 278, 23432-40
  23. Yuan Z, Wang F , Zhao ZW, et al (2011). BIM-Mediated AKT Phosphory-lation Is a Key Modulator of Arsenic Trioxide-Induced Apoptosis in Cisplatin-Sensitive and-Resistant Ovarian Cancer Cells. PLOS ONE, 6, e20586.
  24. Yang JW, Rodrigo R, Felipo V and Lubec GJ (2005), Proteome analysis of primary neurons and astrocytes from rat cerebellum. J Proteome Res, 4, 768-88.
  25. Yan XD, Pan LY, Yuan Y, et al (2007). Identification of platinumresistance associated proteins through proteomic analysis of human ovarian cancer cells and their platinum-resistant sublines. J Proteome Res, 6, 772-80.
  26. Zhou YZ, Chen HZ and Yang QY (1996). Establishment of human ovarian cancer cisplatin resistant cell line COC1/DDP and its mechanism of resistance. Natl Med J China, 76, 680-3.
  27. Zhu BB, Fukada K, Zhu HN and Kyprianou N (2006). Prohibitin and cofilin are intracellular effectors of transforming growth factor beta signaling in human prostate cancer cells. Cancer Res, 66, 8640-7.

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