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Erschienen in: Medical Oncology 3/2012

01.09.2012 | Original Paper

Proteome analysis of the effects of sorafenib on human hepatocellular carcinoma cell line HepG2

verfasst von: Aili Suo, Mingxin Zhang, Yu Yao, Lingmin Zhang, Chen Huang, Kejun Nan, Wanggang Zhang

Erschienen in: Medical Oncology | Ausgabe 3/2012

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Abstract

Sorafenib is a multi-target oral anticancer drug used as first-line treatment for patients with advanced human hepatocellular carcinoma (HCC). But the exact mechanism of sorafenib involved in HCC treatment is not clear yet. In this study, a comparative proteomic approach was performed to identify novel sorafenib-related proteins in HCC. Proteomes of HepG2 cells treated with sorafenib and the control (without sorafenib) were obtained by two-dimensional differential gel electrophoresis. Comprehensive analysis of proteins was focused on total protein spots to filtrate the different protein spots between the two groups. The differentially expressed proteins were identified by peptide mass fingerprinting with high-performance liquid chromatography–tandem mass spectrometry. Then, Western blot and immunohistochemistry were used to verify the expression of some candidate proteins. Results indicated that 19 protein spots were differentially expressed with significant changes, including 6 up-regulated proteins and 13 down-regulated proteins. It was confirmed by Western blot that expressions of Annexin A1 and cyclophilin A were down-regulated in sorafenib-treated HCC cell lines. Immunohistochemical study revealed their oncogenic role in HCC tissues. These observations might be novel findings leading to bring new insights into the exact mechanism of sorafenib and identify possible therapeutic targets.
Literatur
1.
Zurück zum Zitat Bosch FX, Ribes J, Díaz M, Cléries R. Primary liver cancer: worldwide incidence and trends. Gastroenterology. 2004;127(5 suppl 1):S5216. Bosch FX, Ribes J, Díaz M, Cléries R. Primary liver cancer: worldwide incidence and trends. Gastroenterology. 2004;127(5 suppl 1):S5216.
2.
Zurück zum Zitat Lai EC, Lau WY. The continuing challenge of hepatic cancer in Asia. Surgeon. 2005;3(3):210–5.PubMedCrossRef Lai EC, Lau WY. The continuing challenge of hepatic cancer in Asia. Surgeon. 2005;3(3):210–5.PubMedCrossRef
3.
Zurück zum Zitat Gschwind A, Fischer OM, Ullrich A. The discovery of receptor tyrosine kinases: targets for cancer therapy. Nat Rev Cancer. 2004;4(5):361–70.PubMedCrossRef Gschwind A, Fischer OM, Ullrich A. The discovery of receptor tyrosine kinases: targets for cancer therapy. Nat Rev Cancer. 2004;4(5):361–70.PubMedCrossRef
4.
Zurück zum Zitat Dal Lago L, D’Hondt V, Awada A. Selected combination therapy with sorafenib: a review of clinical data and perspectives in advanced solid tumors. Oncologist. 2008;13(8):845–58.PubMedCrossRef Dal Lago L, D’Hondt V, Awada A. Selected combination therapy with sorafenib: a review of clinical data and perspectives in advanced solid tumors. Oncologist. 2008;13(8):845–58.PubMedCrossRef
5.
Zurück zum Zitat Wong SC, Chan CM, Ma BB, Lam MY, Choi GC, et al. Advanced proteomic technologies for cancer biomarker discovery. Expert Rev Proteomics. 2009;6(2):123–34.PubMedCrossRef Wong SC, Chan CM, Ma BB, Lam MY, Choi GC, et al. Advanced proteomic technologies for cancer biomarker discovery. Expert Rev Proteomics. 2009;6(2):123–34.PubMedCrossRef
6.
Zurück zum Zitat Wosikowski K, Regis JT, Robey RW, Alvarez M, Buters JT, et al. Normal p53 status and function despite the development of drug resistance in human breast cancer cells. Cell Growth Differ. 1995;6(11):1395–403.PubMed Wosikowski K, Regis JT, Robey RW, Alvarez M, Buters JT, et al. Normal p53 status and function despite the development of drug resistance in human breast cancer cells. Cell Growth Differ. 1995;6(11):1395–403.PubMed
7.
Zurück zum Zitat Mortz E, Krogh TN, Vorum H, Görg A. Improved silver staining protocols for high sensitivity protein identification using matrix-assisted laser desorption/ionization-time of flight analysis. Proteomics. 2001;1(11):1359–63.PubMedCrossRef Mortz E, Krogh TN, Vorum H, Görg A. Improved silver staining protocols for high sensitivity protein identification using matrix-assisted laser desorption/ionization-time of flight analysis. Proteomics. 2001;1(11):1359–63.PubMedCrossRef
8.
Zurück zum Zitat Andon NL, Hollingworth S, Koller A, Greenland AJ, Yates JR III, et al. Proteomic characterization of wheat amyloplasts using identification of proteins by tandem mass spectrometry. Proteomics. 2002;2(9):1156–68.PubMedCrossRef Andon NL, Hollingworth S, Koller A, Greenland AJ, Yates JR III, et al. Proteomic characterization of wheat amyloplasts using identification of proteins by tandem mass spectrometry. Proteomics. 2002;2(9):1156–68.PubMedCrossRef
9.
Zurück zum Zitat Yates JR III, Eng JK, McCormack AL, Schieltz D. Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database. Anal Chem. 1995;67(8):1426–36.PubMedCrossRef Yates JR III, Eng JK, McCormack AL, Schieltz D. Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database. Anal Chem. 1995;67(8):1426–36.PubMedCrossRef
10.
Zurück zum Zitat Chen HJ, Xiao JR, Yuan W. Loss of p16INK4, alone and with overexpression of osteopontin, correlates with survival of patients with spinal metastasis from hepatocellular carcinoma. Med Oncol. 2010;27(3):1005–9.PubMedCrossRef Chen HJ, Xiao JR, Yuan W. Loss of p16INK4, alone and with overexpression of osteopontin, correlates with survival of patients with spinal metastasis from hepatocellular carcinoma. Med Oncol. 2010;27(3):1005–9.PubMedCrossRef
11.
Zurück zum Zitat Wang J, Zhang M, Zhang L, Cai H, Zhou S, et al. Correlation of Nrf2, HO-1, and MRP3 in gallbladder cancer and their relationships to clinicopathologic features and survival. J Surg Res. 2010;164(1):e99–105.PubMedCrossRef Wang J, Zhang M, Zhang L, Cai H, Zhou S, et al. Correlation of Nrf2, HO-1, and MRP3 in gallbladder cancer and their relationships to clinicopathologic features and survival. J Surg Res. 2010;164(1):e99–105.PubMedCrossRef
12.
Zurück zum Zitat Liu L, Cao Y, Chen C, Zhang X, McNabola A, et al. Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res. 2006;66(24):11851–8.PubMedCrossRef Liu L, Cao Y, Chen C, Zhang X, McNabola A, et al. Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res. 2006;66(24):11851–8.PubMedCrossRef
13.
Zurück zum Zitat Yang Y, Liu X, Fang X, Wang X. Proteomic analysis of mitochondria in Raji cells following exposure to radiation: implications for radiotherapy response. Protein Pept Lett. 2009;16(11):1350–9.CrossRef Yang Y, Liu X, Fang X, Wang X. Proteomic analysis of mitochondria in Raji cells following exposure to radiation: implications for radiotherapy response. Protein Pept Lett. 2009;16(11):1350–9.CrossRef
14.
Zurück zum Zitat Wang W, Goswami S, Lapidus K, Wells AL, Wyckoff JB, et al. Identification and testing of a gene expression signature of invasive carcinoma cells within primary mammary tumors. Cancer Res. 2004;64(23):8585–94.PubMedCrossRef Wang W, Goswami S, Lapidus K, Wells AL, Wyckoff JB, et al. Identification and testing of a gene expression signature of invasive carcinoma cells within primary mammary tumors. Cancer Res. 2004;64(23):8585–94.PubMedCrossRef
15.
Zurück zum Zitat Yanagawa T, Iwasa S, Ishii T, Tabuchi K, Yusa H, et al. Peroxiredoxin I expression in oral cancer: a potential new tumor marker. Cancer Lett. 2000;156(1):27–35.PubMedCrossRef Yanagawa T, Iwasa S, Ishii T, Tabuchi K, Yusa H, et al. Peroxiredoxin I expression in oral cancer: a potential new tumor marker. Cancer Lett. 2000;156(1):27–35.PubMedCrossRef
16.
Zurück zum Zitat Noh DY, Ahn SJ, Lee RA, Kim SW, Park IA, et al. Overexpression of peroxiredoxin in human breast cancer. Anticancer Res. 2001;21(3B):2085–90.PubMed Noh DY, Ahn SJ, Lee RA, Kim SW, Park IA, et al. Overexpression of peroxiredoxin in human breast cancer. Anticancer Res. 2001;21(3B):2085–90.PubMed
17.
Zurück zum Zitat Cha MK, Suh KH, Kim IH. Overexpression of peroxiredoxin I and thioredoxin 1 in human breast carcinoma. J Exp Clin Cancer Res. 2009;28:93.PubMedCrossRef Cha MK, Suh KH, Kim IH. Overexpression of peroxiredoxin I and thioredoxin 1 in human breast carcinoma. J Exp Clin Cancer Res. 2009;28:93.PubMedCrossRef
18.
Zurück zum Zitat Masaki T, Tokuda M, Ohnishi M, Watanabe S, Fujimura T, et al. Enhanced expression of the protein kinase substrate annexin in human hepatocellular carcinoma. Hepatology. 1996;24(1):72–81.PubMed Masaki T, Tokuda M, Ohnishi M, Watanabe S, Fujimura T, et al. Enhanced expression of the protein kinase substrate annexin in human hepatocellular carcinoma. Hepatology. 1996;24(1):72–81.PubMed
19.
Zurück zum Zitat Bai XF, Ni XG, Zhao P, Liu SM, Wang HX, et al. Overexpression of annexin 1 in pancreatic cancer and its clinical significance. World J Gastroenterol. 2004;10(10):1466–70.PubMed Bai XF, Ni XG, Zhao P, Liu SM, Wang HX, et al. Overexpression of annexin 1 in pancreatic cancer and its clinical significance. World J Gastroenterol. 2004;10(10):1466–70.PubMed
20.
Zurück zum Zitat Ahn SH, Sawada H, Ro JY, Nicolson GL. Differential expression of annexin I in human mammary ductal epithelial cells in normal and benign and malignant breast tissues. Clin Exp Metastasis. 1997;15(2):151–6.PubMedCrossRef Ahn SH, Sawada H, Ro JY, Nicolson GL. Differential expression of annexin I in human mammary ductal epithelial cells in normal and benign and malignant breast tissues. Clin Exp Metastasis. 1997;15(2):151–6.PubMedCrossRef
21.
Zurück zum Zitat de Coupade C, Gillet R, Bennoun M, Briand P, Russo-Marie F, et al. Annexin 1 expression and phosphorylation are upregulated during liver regeneration and transformation in antithrombin III SV40 T large antigen transgenic mice. Hepatology. 2000;31(2):371–80.PubMedCrossRef de Coupade C, Gillet R, Bennoun M, Briand P, Russo-Marie F, et al. Annexin 1 expression and phosphorylation are upregulated during liver regeneration and transformation in antithrombin III SV40 T large antigen transgenic mice. Hepatology. 2000;31(2):371–80.PubMedCrossRef
22.
Zurück zum Zitat Campa MJ, Wang MZ, Howard B, Fitzgerald MC, Patz EF Jr. Protein expression profiling identifies macrophage migration inhibitory factor and cyclophilin A as potential molecular targets in non-small cell lung. Cancer Res. 2003;63(7):1652–6.PubMed Campa MJ, Wang MZ, Howard B, Fitzgerald MC, Patz EF Jr. Protein expression profiling identifies macrophage migration inhibitory factor and cyclophilin A as potential molecular targets in non-small cell lung. Cancer Res. 2003;63(7):1652–6.PubMed
23.
Zurück zum Zitat Li M, Zhai Q, Bharadwaj U, Wang H, Li F, et al. Cyclophilin A is overexpressed in human pancreatic cancer cells and stimulates cell proliferation through CD147. Cancer. 2006;106(10):2284–94.PubMedCrossRef Li M, Zhai Q, Bharadwaj U, Wang H, Li F, et al. Cyclophilin A is overexpressed in human pancreatic cancer cells and stimulates cell proliferation through CD147. Cancer. 2006;106(10):2284–94.PubMedCrossRef
24.
Zurück zum Zitat Zheng J, Koblinski JE, Dutson LV, Feeney YB, Clevenger CV. Prolyl isomerase cyclophilin A regulation of Janus-activated kinase 2 and the progression of human breast cancer. Cancer Res. 2008;68(19):7769–78.PubMedCrossRef Zheng J, Koblinski JE, Dutson LV, Feeney YB, Clevenger CV. Prolyl isomerase cyclophilin A regulation of Janus-activated kinase 2 and the progression of human breast cancer. Cancer Res. 2008;68(19):7769–78.PubMedCrossRef
25.
Zurück zum Zitat Wong CS, Wong VW, Chan CM, Ma BB, Hui EP, et al. Identification of 5-fluorouracil response proteins in colorectal carcinoma cell line SW480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Oncol Rep. 2008;20(1):89–98.PubMed Wong CS, Wong VW, Chan CM, Ma BB, Hui EP, et al. Identification of 5-fluorouracil response proteins in colorectal carcinoma cell line SW480 by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Oncol Rep. 2008;20(1):89–98.PubMed
26.
Zurück zum Zitat Huang CF, Sun ZJ, Zhao YF, Chen XM, Jia J, et al. Increased expression of peroxiredoxin 6 and cyclophilin A in squamous cell carcinoma of the tongue. Oral Dis. 2011;17(3):328–34.PubMedCrossRef Huang CF, Sun ZJ, Zhao YF, Chen XM, Jia J, et al. Increased expression of peroxiredoxin 6 and cyclophilin A in squamous cell carcinoma of the tongue. Oral Dis. 2011;17(3):328–34.PubMedCrossRef
27.
Zurück zum Zitat Al-Ghoul M, Brück TB, Lauer-Fields JL, Asirvatham VS, Zapata C, et al. Comparative proteomic analysis of matched primary and metastatic melanoma cell lines. J Proteome Res. 2008;7(9):4107–18.PubMedCrossRef Al-Ghoul M, Brück TB, Lauer-Fields JL, Asirvatham VS, Zapata C, et al. Comparative proteomic analysis of matched primary and metastatic melanoma cell lines. J Proteome Res. 2008;7(9):4107–18.PubMedCrossRef
28.
Zurück zum Zitat Lee J. Novel combinational treatment of cisplatin with cyclophilin A inhibitors in human hepatocellular carcinomas. Arch Pharm Res. 2010;33(9):1401–9.PubMedCrossRef Lee J. Novel combinational treatment of cisplatin with cyclophilin A inhibitors in human hepatocellular carcinomas. Arch Pharm Res. 2010;33(9):1401–9.PubMedCrossRef
29.
Zurück zum Zitat Dai Z, Liu YK, Cui JF, Shen HL, Chen J, et al. Identification and analysis of altered alpha1, 6-fucosylated glycoproteins associated with hepatocellular carcinoma metastasis. Proteomics. 2006;6(21):5857–67.PubMedCrossRef Dai Z, Liu YK, Cui JF, Shen HL, Chen J, et al. Identification and analysis of altered alpha1, 6-fucosylated glycoproteins associated with hepatocellular carcinoma metastasis. Proteomics. 2006;6(21):5857–67.PubMedCrossRef
30.
Zurück zum Zitat Goan YG, Zhou B, Hu E, Mi S, Yen Y. Overexpression of ribonucleotide reductase as a mechanism of resistance to 2, 2-difluorodeoxycytidine in the human KB cancer cell line. Cancer Res. 1999;59(17):4204–7.PubMed Goan YG, Zhou B, Hu E, Mi S, Yen Y. Overexpression of ribonucleotide reductase as a mechanism of resistance to 2, 2-difluorodeoxycytidine in the human KB cancer cell line. Cancer Res. 1999;59(17):4204–7.PubMed
31.
Zurück zum Zitat Cao MY, Lee Y, Feng NP, Xiong K, Jin H, et al. Adenovirus-mediated ribonucleotide reductase R1 gene therapy of human colon adenocarcinoma. Clin Cancer Res. 2003;9(12):4553–61.PubMed Cao MY, Lee Y, Feng NP, Xiong K, Jin H, et al. Adenovirus-mediated ribonucleotide reductase R1 gene therapy of human colon adenocarcinoma. Clin Cancer Res. 2003;9(12):4553–61.PubMed
32.
Zurück zum Zitat Davidson JD, Ma L, Flagella M, Geeganage S, Gelbert LM, et al. An increase in the expression of ribonucleotide reductase large subunit 1 is associated with gemcitabine resistance in non-small cell lung cancer cell lines. Cancer Res. 2004;64(11):3761–6.PubMedCrossRef Davidson JD, Ma L, Flagella M, Geeganage S, Gelbert LM, et al. An increase in the expression of ribonucleotide reductase large subunit 1 is associated with gemcitabine resistance in non-small cell lung cancer cell lines. Cancer Res. 2004;64(11):3761–6.PubMedCrossRef
33.
Zurück zum Zitat Rosell R, Danenberg KD, Alberola V, Bepler G, Sanchez JJ, et al. Ribonucleotide reductase messenger RNA expression and survival in gemcitabine/cisplatin-treated advanced non-small cell lung cancer patients. Clin Cancer Res. 2004;10(4):1318–25.PubMedCrossRef Rosell R, Danenberg KD, Alberola V, Bepler G, Sanchez JJ, et al. Ribonucleotide reductase messenger RNA expression and survival in gemcitabine/cisplatin-treated advanced non-small cell lung cancer patients. Clin Cancer Res. 2004;10(4):1318–25.PubMedCrossRef
34.
Zurück zum Zitat Phadke MS, Krynetskaia NF, Mishra AK, Krynetskiy E. Glyceraldehyde 3-phosphate dehydrogenase depletion induces cell cycle arrest and resistance to antimetabolites in human carcinoma cell lines. J Pharmacol Exp Ther. 2009;331(1):77–86.PubMedCrossRef Phadke MS, Krynetskaia NF, Mishra AK, Krynetskiy E. Glyceraldehyde 3-phosphate dehydrogenase depletion induces cell cycle arrest and resistance to antimetabolites in human carcinoma cell lines. J Pharmacol Exp Ther. 2009;331(1):77–86.PubMedCrossRef
35.
Zurück zum Zitat Ménasché G, Pastural E, Feldmann J, Certain S, Ersoy F, et al. Mutations in RAB27A cause Griscelli syndrome associated with haemophagocytic syndrome. Nat Genet. 2000;25(2):173–6.PubMedCrossRef Ménasché G, Pastural E, Feldmann J, Certain S, Ersoy F, et al. Mutations in RAB27A cause Griscelli syndrome associated with haemophagocytic syndrome. Nat Genet. 2000;25(2):173–6.PubMedCrossRef
36.
Zurück zum Zitat Wang JS, Wang FB, Zhang QG, Shen ZZ, Shao ZM. Enhanced expression of Rab27A gene by breast cancer cells promoting invasiveness and the metastasis potential by secretion of insulin-like growth factor-II. Mol Cancer Res. 2008;6(3):372–82.PubMedCrossRef Wang JS, Wang FB, Zhang QG, Shen ZZ, Shao ZM. Enhanced expression of Rab27A gene by breast cancer cells promoting invasiveness and the metastasis potential by secretion of insulin-like growth factor-II. Mol Cancer Res. 2008;6(3):372–82.PubMedCrossRef
37.
Zurück zum Zitat Gatenby RA, Gillies RJ. Glycolysis in cancer: a potential target for therapy. Int J Biochem Cell Biol. 2007;39(7–8):1358–66.PubMedCrossRef Gatenby RA, Gillies RJ. Glycolysis in cancer: a potential target for therapy. Int J Biochem Cell Biol. 2007;39(7–8):1358–66.PubMedCrossRef
38.
Zurück zum Zitat Fiume L, Vettraino M, Manerba M, Di Stefano G. Inhibition of lactic dehydrogenase as a way to increase the anti-proliferative effect of multi-targeted kinase inhibitors. Pharmacol Res. 2011;63(4):328–34.PubMedCrossRef Fiume L, Vettraino M, Manerba M, Di Stefano G. Inhibition of lactic dehydrogenase as a way to increase the anti-proliferative effect of multi-targeted kinase inhibitors. Pharmacol Res. 2011;63(4):328–34.PubMedCrossRef
Metadaten
Titel
Proteome analysis of the effects of sorafenib on human hepatocellular carcinoma cell line HepG2
verfasst von
Aili Suo
Mingxin Zhang
Yu Yao
Lingmin Zhang
Chen Huang
Kejun Nan
Wanggang Zhang
Publikationsdatum
01.09.2012
Verlag
Springer US
Erschienen in
Medical Oncology / Ausgabe 3/2012
Print ISSN: 1357-0560
Elektronische ISSN: 1559-131X
DOI
https://doi.org/10.1007/s12032-011-0013-y

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