Lipocalin: A Novel Diagnostic Marker for Hepatocellular Carcinoma in Chronic Liver Disease Patients in Egypt

Hoda Aly Abd El Moety, Rania Mahamed El Sharkawy, Nevin Abd El Menem Hussein

Chemical Pathology, Medical Research Institute, Alexandria University, Alexandria, Egypt.
Chemical Pathology, Medical Research Institute, Alexandria University, Alexandria, Egypt;.
Chemistry, Medical Research Institute, Alexandria University, Alexandria, Egypt..

DOI: 10.4236/ijcm.2013.410079                

Abstract

Hepatocellular carcinoma is one of the most prevalent life-threatening human cancers with etiological factors chronic viral hepatitis B and C infections. Tumor cell dispersion relies on the loss of homotypic cell-cell adhesion. Invasion through basement membrane and interstitial extracellular matrix is another key event for metastatic progression, which requires the action of a series of proteolytic enzymes named matrix metalloproteinases, secreted by tumor cells that enhance tumor invasiveness and metastasis. TIMPs are dominant inhibitors of MMPs and able to control MMP-mediated ECM breakdown by binding active forms of MMPs. Lipocalin-2 is known as neutrophil gelatinase associated lipocalin promotematrix degradation and tumor progression. Aim: To evaluate the importance of lipocalin for diagnosis HCC in Egyptian chronic liver disease patients. Subjects and Methods: 50 patients and 25 controls. (G-1) 25 HCC on top of hepatitis C. (G-2) 25 hepatitis C. The following done: schistosoma antibodies, ASAM, LKM-1, ANA AKA and CBC. Hepatitis B surface antigen, Hepatitis C antibodies AFP, Cupper and zinc, Matrix metaloprotinase-9, TIMP-1 and Neutrophil gelatinase-associated lipocalin. Results: Median value of MMP-9 level in G-I (206 μg/L) is significantly higher than G-2 (100 μg/L) and G 3 (49 μg/L) p < 0.001. TIMP-9 median value G-1 (48 μg/L) is significantly lower than G-2 (54 μg/L) and G-3 (113 μg/L) p < 0.001. Lipocalin-2 median levels are significantly higher in G-I (389 ng/mL) than G-2 (166 ng/ml) versus G-3 (60 ng/mL) p < 0.001. Lipocalin-2 associated with increasing lobular inflammation, ballooning & fibrosis with MMP-9 has an important role in pathogenesis of liver cirrhosis and HCC. Conclusion: We elucidated predictive value for MMPs, TIMPs, and progression metastasis of hepatocellular carcinoma. Liver cell impairment alters metabolism of trace metals as zinc and copper, with possible relationship of these changes to pathogenesis of chronic liver disease. Lipocalin-2 can be used as a future diagnostic marker with better sensitivity and specificity than MMP-9 for the progression of hepatocelluar carcinoma.

Keywords

Lipocalin-2; MMP-9; TIMP-1; Cu; Zn; HCC

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	[1] M. M. Center and A. Jemal, “International Trends in Liver Cancer Incidence Rates,” Cancer Epidemiology, Biomarkers & Prevention, Vol. 20, No. 11, 2011, pp. 2362-2368. http://dx.doi.org/10.1158/1055-9965.EPI-11-0643
[2]	E. P. Simard, E. M. Ward, R. Siegel and A. Jemal, “Cancers with Increasing Incidence Trends in the United States 1999 through 2008,” CA: A Cancer Journal for Clinicians, 2012, in press.
[3]	S. S. Thorgeirsson and J. W. Grisham, “Molecular Pathogenesis of Human Hepatocellular Carcinoma,” Nature Genetics, Vol. 31, No. 4, 2002, pp. 339-346. http://dx.doi.org/10.1038/ng0802-339
[4]	P. A. Farazi and R. A. de Pinho, “Hepatocellular Carcinoma Pathogenesis: From Genes to Environment,” Nature Reviews Cancer, Vol. 6, No. 9, 2006, pp. 674-687. http://dx.doi.org/10.1038/nrc1934
[5]	A. P. Venook, C. Papandreou, J. Furuse and L. L. de Guevara, “The Incidence and Epidemiology of Hepatocellular Carcinoma: A Global and Regional Perspective,” Oncologist, Vol. 15, No. 4, 2010, pp. 5-13. http://dx.doi.org/10.1634/theoncologist.2010-S4-05
[6]	J. D. Yang and L. R. Roberts, “Epidemiology and Management of Hepatocellular Carcinoma,” Infectious Disease Clinics of North America, Vol. 24, No. 4, 2010, pp. 899-919. http://dx.doi.org/10.1016/j.idc.2010.07.004
[7]	D. Kremsdorf, P. Soussan, P. Paterlini-Brechot and C. Brechot, “Hepatitis B Virus Related Hepatocellular Carcinoma: Paradigms for Viral-Related Human Carcinogenesis,” Oncogene, Vol. 25, No. 27, 2006, pp. 3823-3833. http://dx.doi.org/10.1038/sj.onc.1209559
[8]	M. Levrero, “Viral Hepatitis and Liver Cancer: The Case of Hepatitis C,” Oncogene, Vol. 25, 2006, pp. 3834-3847. http://dx.doi.org/10.1038/sj.onc.1209562
[9]	J. A. Sanyal, S. K. Yoon and R. Lencioni, “The Etiology of Hepatocellular Carcinoma and Consequences for Treatment,” Oncologist, Vol. 15, No. 4, 2010, pp. 14-22. http://dx.doi.org/10.1634/theoncologist.2010-S4-14
[10]	A. B. Siegel and A. X. Zhu, “Metabolic Syndrome and Hepatocellular Carcinoma,” Cancer, Vol. 115, No. 24, 2009, pp. 5651-5661. http://dx.doi.org/10.1002/cncr.24687
[11]	T. M. Welzel, B. I. Graubard, S. Zeuzem, H. B. El-Serag, J. A. Davila and K. A. McGlynn, “Metabolic Syndrome Increases Risk of Primary Liver Cancer in the United States: A Study in the SEER-Medicare Database,” Hepatology, Vol. 54, No. 2, 2011, pp. 463-471. http://dx.doi.org/10.1002/hep.24397
[12]	E. Hashimoto and K. Tokushige, “Hepatocellular Carcinoma in Non-Alcoholic Steatohepatitis: Growing Evidence of an Epidemic,” Hepatology Research, Vol. 42, No. 1, 2012, pp. 1-14. http://dx.doi.org/10.1111/j.1872-034X.2011.00872.x
[13]	L. Libbrecht, V. Desmet and T. Roskams, “Preneoplastic Lesions in Human Hepatocarcinogenesis,” Liver International, Vol. 25, No. 1, 2005, pp. 16-27. http://dx.doi.org/10.1111/j.1478-3231.2005.01016.x
[14]	B. P. L. Wijnhoven, W. N. M. Dinjens and M. Pignatelli, “Ecadherin-Catenin Cell-Cell Adhesion Complex and Human Cancer,” British Journal of Surgery, Vol. 87, No. 8, 2000, pp. 992-1005. http://dx.doi.org/10.1046/j.1365-2168.2000.01513.x
[15]	H. Brikedal-hansen, W. G. I. Moore, M. K. Bodden, et al., “Matrix Metalloproteinases: A Review,” Critical Reviews in Oral Biology & Medicine, Vol. 4, No. 2, 1993, pp. 197-250.
[16]	M. Yoshimoto, F. Itoh, H. Yamamoto, et al., “Expression of MMP-7 (Pump-1) mRNA in Human Colorectal Cancers,” International Journal of Cancer, Vol. 54, No. 4, 1993, pp. 614-618. http://dx.doi.org/10.1002/ijc.2910540415
[17]	A. Senota, F. Itoh, H. Yamamoto, et al., “Relation of Matrilysin Messenger RNA Expression with Invasive Activity in Human Gastric Cancer,” Clinical and Experimental Metastasis, Vol. 16, No. 4, 1998, pp. 313-321. http://dx.doi.org/10.1023/A:1006509312674
[18]	A. J. Docherty, J. O’Conell, T. Crabbe, et al., “The Matrix Metalloproteinases and Their Natural Inhibitors: Prospects for Treating Degenerative Tissue Diseases,” Trends in Biotechnology, Vol. 10, 1992, pp. 200-207. http://dx.doi.org/10.1016/0167-7799(92)90214-G
[19]	L. Matrisian, “Metalloproteinases and Their Inhibitors in Matrix Remodeling,” Trends in Genetics, Vol. 6, 1990, pp. 121-125. http://dx.doi.org/10.1016/0168-9525(90)90126-Q
[20]	L. A. Liotta, “Tumor Invasion and Metastases-Role of the Extracellular Matrix: Rhoads Memorial Award Lecture,” Cancer Research, Vol. 46, No. 1, 1986, pp. 1-7.
[21]	L. A. Liotta and E. C. Kohn, “The Microenvironment of the Tumour-Host Interface,” Nature, Vol. 411, No. 6835, 2001, pp. 375-379. http://dx.doi.org/10.1038/35077241
[22]	P. M. Comoglio and L. Trusolino, “Invasive Growth: From Development to Metastasis,” Journal of Clinical Investigation, Vol. 109, No. 7, 2002, pp. 857-862.
[23]	L. M. Coussens and Z. Werb, “Matrix Metalloproteinases and the Development of Cancer,” Chemistry & Biology, Vol. 3, No. 11, 1996, pp. 895-904. http://dx.doi.org/10.1016/S1074-5521(96)90178-7
[24]	L. E. Littlepage, M. Egeblad and Z. Werb, “Coevolution of Cancer and Stromal Cellular Responses,” Cancer Cell, Vol. 7, No. 6, 2005, pp. 499-500. http://dx.doi.org/10.1016/j.ccr.2005.05.019
[25]	R. Lichtinghagen, T. Helmbrecht, B. Arndt, et al., “Expression Pattern of Matrix Metalloproteinases in Human Liver,” European Journal of Clinical Chemistry and Clinical Biochemistry, Vol. 33, 1995, pp. 65-71.
[26]	A. H. Baker, D. R. Edwards and G. Murphy, “Metalloproteinase Inhibitors: Biological Actions and Therapeutic Opportunities,” Journal of Cell Science, Vol. 115, No. 19, 2002, pp. 3719-3727. http://dx.doi.org/10.1242/jcs.00063
[27]	T. E. Curry Jr. and K. G. Osteen, “The Matrix Metalloproteinase System: Changes, Regulation, and Impact through the Ovarian and Uterine Reproductive Cycle,” Endocrine Review, Vol. 24, No. 4, 2003, pp. 428-465. http://dx.doi.org/10.1210/er.2002-0005
[28]	C. M. Dollery, J. R. McEwan and A. M. Henney, “Matrix Metalloproteinases and Cardiovascular Disease,” Circulation Research, Vol. 77, No. 5, 1995, pp. 863-868. http://dx.doi.org/10.1161/01.RES.77.5.863
[29]	S. F. Yan, M. K. Chen, Y. S. Hsieh, et al., “Antimetastatic of Terminalia catappa L on Oral Cancer via Down Regulation of Metastasis-Associated Proteses,” Food Chemical Toxicology, Vol. 48, No. 4, 2012, pp. 1052-1058.
[30]	C. B. Yeh, M. J. Hsieh, Y. H. Hseih, et al., “Antimetastatic Effects of Norcantharidin on Hepatocellular Carcinoma by Transcriptional Inhibition of MMP-9 through Modulation of NF-kB Activity,” Plos ONE, Vol. 45, 2012, pp. 234-236.
[31]	C. B. Yeh, M. J. Hsieh, Y. S. Hsieh, M. H. Chien, P. Y. Lin, H. L. Chiou, et al., “Terminalia Catappa Exerts Antimetastatic Effects on Hepatocellular Carcinoma through Transcriptional Inhibition of Matrix Metalloproteinase-9 by Modulating NF-κB and AP-1 Activity,” EvidenceBased Complementary and Alternative Medicine, Vol. 59, 2012, pp. 52-92.
[32]	R. Parkash and P. John, “Macrophages: Central Regulator of Hepatic Fibrogenesis and Fibrosis Resolution,” Journal of Hepatology, Vol. 56, No. 6, 2012, pp. 1417-1419. http://dx.doi.org/10.1016/j.jhep.2011.10.026
[33]	B. Marina and D. Schuppan, “Progression of Liver Fibrosis in Post-Transplant Hepatitis C: Mechanism, Assessment and Treatment,” Journal of Hepatology, Vol. 58, No. 5, 2013, pp. 1028-1041. http://dx.doi.org/10.1016/j.jhep.2012.12.014
[34]	L. Yan, N. Borregaard, L. Kjeldsen and M. A. Moses, “The High Molecular Weight Urinary Matrix Metalloproteinase (MMP) Activity Is a Complex of Gelatinase B/ MMP-9 and Neutrophil Gelatinase-Associated Lipocalin (NGAL). Modulation of MMP-9 Activity by NGAL,” Journal of Biological Chemistry, Vol. 276, No. 40, 2001, pp. 37258-37265. http://dx.doi.org/10.1074/jbc.M106089200
[35]	R. Roy, G. Louis, K. R. Loughlin, D. Wiederschain, S. M. Kilroy, C. C. Lamb, et al., “Tumor-Specific Urinary Matrix Metalloproteinase Fingerprinting: Identification of High Molecular Weight Urinary Matrix Metalloproteinase Species,” Clinical Cancer Research, Vol. 14, No. 20, 2008, pp. 6610-6617. http://dx.doi.org/10.1158/1078-0432.CCR-08-1136
[36]	C. A. Fernandez, L. Yan, G. Louis, J. Yang, J. L. Kutok and M. A. Moses, “The Matrix Metalloproteinase-9/Neutrophil Gelatinase-Associated Lipocalin Complex Plays a Role in Breast Tumor Growth and Is Present in the Urine of Breast Cancer Patients,” Clinical Caner Research, Vol. 11, 2005, pp. 5390-5395. http://dx.doi.org/10.1158/1078-0432.CCR-04-2391
[37]	M. E. Azab and E. A. El Zayat, “Evaluation of Purified Antigens in Haemagglutination Test (IHA) for Determination of Cross Reactivates in Diagnosis of Fascioliasis and Schistosomiasis,” Journal of the Egyptian Society of Parasitology, Vol. 26, No. 3, 1996, pp. 677-685.
[38]	A. Radice, L. Bianchi and R. A. Sinico, “Anti-Neutrophil Cytoplasmic Autoantibodies: Methodological Aspects and Clinical Significance in Systemic Vasculitis,” Autoimmunity Reviews, Vol. 12, No. 4, 2013, pp. 487-495. http://dx.doi.org/10.1016/j.autrev.2012.08.008
[39]	B. Bain, S. Lewis and I. Bates, “Basic Haematological Techniques,” In: S. Lewis, B. Bain and I. Bates, Eds., Practical Haematology, Churchill Livingstone, Philadelphia, 2006, pp. 25-58.
[40]	G. Walters, L. P. Kuijperst, J. Kaccaki and A. H. Schuurs, “Enzymes Linked Immunosorbent Assay for Hepatitis B Surface Antigen,” The Journal of Infectious Disease, Vol. 136, Suppl. 2, 1997, pp. S311-S317. http://dx.doi.org/10.1093/infdis/136.Supplement_2.S311
[41]	C. A. Burtis, E. R. Ashwood and D. E. Bruns, “Tietz Textbook of Clinical Chemistry and Molecular Diagnostics,” 4th Edition, Elsevier Saunders Company, St Louis, 2006.
[42]	T. Makimo and K. Takahara, “Determination of Copper and Zinc in Infants by Atomic Absorption with Discrete Nebulization,” Clinical Chemistry, Vol. 27, No. 8, 1981, pp. 1445-1447.
[43]	C. Jumper, E. Cobos and C. Lox, “Determination of the Serum Matrix Metalloproteinase-9 (MMP-9) and Tissue Inhibitor of Matrix Metalloproteinase-1 (TIMP-1) in Patients with Either Advanced Small-Cell Lung Cancer or Non-Small-Cell Lung Cancer Prior to Treatment,” Respiratory Medicine, Vol. 98, No. 2, 2004, pp. 173-177. http://dx.doi.org/10.1016/j.rmed.2003.08.014
[44]	I. Toshihiko, F. Takehiko and S. Makoto, “Elevated Levels of Circulating Plasma Matrix Metalloproteinase 9 in Non-Small Cell Lung Cancer Patients,” Clinical Cancer Research, Vol. 5, No. 1, 1999, pp. 149-153.
[45]	K. R. Pedersen, H. B. Ravn, V. E. Hjortdal, R. Narregaard and J. V. Povlsen, “Neutrophil Gelatinase-Associated Lipocalin (NGAL): Validation of Commercially Available ELISA,” Scandinavian Journal of Clinical and Laboratory Investigation, Vol. 70, No. 5, 2010, pp. 374-382. http://dx.doi.org/10.3109/00365513.2010.486868
[46]	B. Gyorgy, M. Elizabeth and H. Stephen, “Hepatocellular Carcinoma in Non-Alcoholic Fatty Liver Disease: An Emerging Menace,” Journal of Hepatology, Vol. 56, No. 6, 2012, pp. 1384-1391. http://dx.doi.org/10.1016/j.jhep.2011.10.027
[47]	A. Jemal, F. Bray, M. M. Center, et al., “Global Cancer Statistics,” CA: A Cancer Journal for Clinicians, Vol. 61, No. 2, 2011, pp. 69-90. http://dx.doi.org/10.3322/caac.20107
[48]	H. B. EL-serag, “Hepatocellular Carcinoma,” New England Journal of Medicine, Vol. 365, No. 12, 2011, pp. 1118-1127.
[49]	F. X. Bosch, J. Ribes, R. Cleries and M. Diaz, “Epidemiology of Hepatocellular Carcinoma,” Clinics in Liver Disease, Vol. 9, No. 2, 2005, pp. 191-211. http://dx.doi.org/10.1016/j.cld.2004.12.009
[50]	J. P. Michael, “Fibrogenesis II. Metalloproteinases and Their Inhibitors in Liver Fibrosis,” American Journal of Physiology-Gastrointestinal and Liver Physiology, Vol. 279, No. 2, 2000, pp. G245-G249.
[51]	R. C. Benyon, C. J. Hovell, M. D. A. Gaca, et al., “Progelatinase A Is Produced and Activated by the Hepatic Stellate Cells and Promotes Their Proliferation,” Hepatology, Vol. 30, No. 4, 1999, pp. 977-986. http://dx.doi.org/10.1002/hep.510300431
[52]	P. J. Winwood, D. Schuppan, J. P. Iredale, et al., “Kupffer Cell-Derived 95kD a Type IV Collagenase/Gelatinase B: Characterisation and Expression in Cultured Cells,” Hepatology, Vol. 22, No. 1, 1995, pp. 304-315.
[53]	H. Leyland, J. Gentry, M. J. P. Arthur and R. C. Benyon, “The Plasminogen-Activating System in Hepatic Stellate Cells,” Hepatology, Vol. 24, No. 5, 1996, pp. 1172-1178. http://dx.doi.org/10.1002/hep.510240532
[54]	N. Theret, K. Lehti, O. Musso and B. Clement, “MMP2 Activation by Collagen I and Concanavalin A in Cultured Human Hepatic Stellate Cells,” Hepatology, Vol. 30, No. 2, 1999, pp. 462-468. http://dx.doi.org/10.1002/hep.510300236
[55]	L. Ralf, H. Omar, S. Thomas, et al., “Expression of Matrix Metalloproteinase-2 and-9 and Their Inhibitors in Peripheral Blood Cells of Patients with Chronic Hepatitis C,” Clinical Chemistry, Vol. 46, No. 2, 2000, pp. 183-192.
[56]	B. Ramón, A. David and D. Brenner, “Liver Fibrosis,” The Journal of Clinical Investigation, Vol. 115, No. 2, 2005, pp. 209-218.
[57]	S. Zhang, L. Li, J. Y. Lin and H. Lin, “Imbalance between Expression of Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-1 in Invasiveness and Metastasis of Human Gastric Carcinoma,” World Journal of Gastroenterology, Vol. 9, No. 5, 2003, pp. 899-904.
[58]	G. Zu, S. Maria, L. Wen, et al., “Association of E-Cadherin, Matrix Metalloproteinases, and Tissue Inhibitors of Metalloproteinases with the Progression and Metastasis of Hepatocellular Carcinoma,” Modern Pathology, Vol. 19, 2006, pp. 533-540. http://dx.doi.org/10.1038/modpathol.3800554
[59]	S. Xu, C. Petersson, M. Carlson and P. Venge, “The Development of an Assay for Human Neutrophil Lipocalin (HNL) to Be Used as a Specific Marker of Neutrophil Activity in Vivo and Vitro,” Journal of Immunological Methods, Vol. 171, No. 2, 1994, pp. 245-252. http://dx.doi.org/10.1016/0022-1759(94)90044-2
[60]	D. R. Flower, “The Lipocalin Protein Family: A Role in Cell Regulation,” FEBS Letters, Vol. 354, No. 1, 1994, pp. 7-11. http://dx.doi.org/10.1016/0014-5793(94)01078-1
[61]	T. Harald, Z. Volker, T. Susanne, et al., “The Human Neutrophil Lipocalin Supports the Allosteric Activation of Matrix Metalloproteinases,” European Journal of Biochemistry, Vol. 268, No. 7, 2001, pp. 1918-1928. http://dx.doi.org/10.1046/j.1432-1327.2001.02066.x
[62]	O. S. Kwon, D. Y. Lim, K. A. Kwon, M. G. Chung, D. K. Park, S. S. Kim, et al., “Clinical Usefulness of Plasma Activities of Gelatinase (Matrix Metalloproteinase-2 and 9) in Chronic Liver Disease,” Taehan Kan Hakhoe Chi, Vol. 9, No. 3, 2003, pp. 222-230.
[63]	K. Suzuki, R. Oyama, E. Hayashi and Y. Arakawa, “Liver Diseases and Essential Trace Elements,” Nihon Rinsho, Vol. 54, No. 1, 1996, pp. 85-92.
[64]	R. Hatano, M. Ebara, H. Fukuda, M. Yoshikawa, et al., “Accumulation of Copper in the Liver and Hepatic Injury in Chronic Hepatitis C,” Journal of Gastroenterology and Hepatology, Vol. 15, No. 7, 2000, pp. 786-791. http://dx.doi.org/10.1046/j.1440-1746.2000.02199.x
[65]	R. Hatano, M. Ebara, H. Fukuda, M. Yoshikawa, N. Sugiura, F. Kondo, et al., “Accumulation of Copper in the Liver and Hepatic Injury in Chronic Hepatitis C,” Journal of Gastroenterology and Hepatology, Vol. 15, No. 7, 2000, pp. 786-791. http://dx.doi.org/10.1046/j.1440-1746.2000.02199.x
[66]	K. Suzuki, R. Oyama, E. Hayashi and Y. Arakawa, “Liver Diseases and Essential Trace Elements,” Nihon Rinsho, Vol. 54, No. 1, 1996, pp. 85-92.
[67]	M. N. Rashed, “The Role of Trace Elements on Hepatitis Virus Infections: A Review,” Journal of Trace Elements in Medicine and Biology, Vol. 25, No. 3, 2011, pp. 181-187. http://dx.doi.org/10.1016/j.jtemb.2011.07.001
[68]	C. Pramoolsinsap, N. Promvanit, S. Komindr, et al., “Serum Trace Metals in Chronic Viral Hepatitis and Hepatocellular Carcinoma in Thailand,” Journal of Gastroenterology, Vol. 29, No. 5, 1994, pp. 610-615. http://dx.doi.org/10.1007/BF02365444