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Studies on Focal Adhesion Kinase in Human Breast Cancer Tissue

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DOI: 10.4236/jct.2012.31002    4,102 Downloads   7,723 Views   Citations

ABSTRACT

Aim: To study Expression and Phosphorylation status of Focal Adhesion Kinase (FAK) in Human Breast Cancer tissue. To study the relation of FAK with standard clinicopathological parameters of Human Breast Cancer. Methods: Tissue collection, Protein extraction, RNA isolation, Western Blot, Immunohistochemistry, RT-PCR, ELISA, Statistical analysis. Results: All the four techniques showed upregulated expression, phosphorylation (Tyr-397) and processing of FAK in human breast cancer tissue compared to the adjacent non-tumor tissue of the same patient. Upregulation of FAK was found to be increased parallely with the advancement of cancer. Localisation of FAK was found to be membrano-cytoplasmic. FAK is upregulated both in protein and mRNA level. Expression and phosphorylation of FAK is increased specifically in the tumor regions compared to the surrounding non-tumor region. Upregulation of FAK was frequently found in ER-positive and PR-positive but Her2/neunegative breast cancer cases. Conclusion: FAK has crucial role in development and progression of human breast cancer. FAK may be considered as an indicator of human breast cancer progression. FAK processing may be considered as an indicator of invasive potential of breast cancer. FAK may be considered as a clinical indicator of human breast cancer development and progression.

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K. Ganguly, T. Sen, S. Mandal, J. Biswas and A. Chatterjee, "Studies on Focal Adhesion Kinase in Human Breast Cancer Tissue," Journal of Cancer Therapy, Vol. 3 No. 1, 2012, pp. 7-19. doi: 10.4236/jct.2012.31002.

References

[1] M. Vicente-Manzanares, D. J. Webb and A. R. Horwitz, “Cell Migration at a Glance,” Journal of Cell Science, Vol. 118, No. 21, 2005, pp. 4917-4919. doi:10.1242/jcs.02662
[2] S. K. Mitra, D. A. Hanson and D. D. Schlaepfer, “Focal Adhesion Kinase: In Command and Control of Cell Motility,” Nature Reviews Molecular Cell Biology, Vol. 6, No. 1, 2005, pp. 56-68. doi:10.1038/nrm1549
[3] D. D. Schlaepfer and S. K. Mitra, “Multiple Connections Link FAK to Cell Motility and Invasion,” Current Opinion in Genetics & Development, Vol. 14, No. 1, 2004, pp. 92-101. doi:10.1016/j.gde.2003.12.002
[4] A. Hamadi, M. Bouali, M. Dontenwill, H. Stoeckel, K. Takeda and P. Ronde, “Regulation of Focal Adhesion Dynamics and Disassembly by Phosphorylation of FAK at Tyrosine 397,” Journal of Cell Science, Vol. 118, No. 19, 2005, pp. 4415-4425. doi:10.1242/jcs.02565
[5] J. D. Owen, P. J. Ruest, D. W. Fry and S. K. Hanks, “Induced Focal Adhesion Kinase (FAK) Expression in FAK-Null Cells Enhances Cell Spreading and Migration Requiring Both Auto- and Activation Loop Phosphorylation Sites and Inhibits Adhesion-Dependent Tyrosine Phosphorylation of Pyk2,” Molecular and Cellular Biology, Vol. 19, No. 7, 1999, pp. 4806-4818.
[6] L. A. Cary, J. F. Chang and J. L. Guan, “Stimulation of Cell Migration by Overexpression of Focal Adhesion Kinase and Its Association with Src and Fyn,” Journal of Cell Science, Vol. 109, No. 7, 1996, pp. 1787-1794.
[7] R. E. Gates, L. E. King Jr., S. K. Hanks and L. B. Nanney, “Potential Role for Focal Adhesion Kinase in Migrating and Proliferating Keratinocytes near Epidermal Wounds and in Culture,” Cell Growth and Differentiation, Vol. 5, No. 8, 1994, pp. 891-899.
[8] D. Ilic, Y. Furuta, S. Kanazawa, N. Takeda, K. Sobue, N. Nakatsuji, S. Nomura, J. Fujimoto, M. Okada and T. Yamamoto, “Reduced Cell Motility and Enhanced Focal Adhesion Contact Formation in Cells from FAK Deficient Mice,” Nature, Vol. 377, No. 6549, 1995, pp. 539-544. doi:10.1038/377539a0
[9] D. J. Sieg, C. R. Hauck and D. D. Schlaepfer, “Required role of Focal Adhesion Kinase (FAK) for Integrin-Stimulated Cell Migration,” Journal of Cell Science, Vol. 112, No. 16, 1999, pp. 2677-2691.
[10] M. A. Westhoff, B. Serrels, V. J. Fincham, M. C. Frame and N. O. Carragher, “SRC-Mediated Phosphorylation of Focal Adhesion Kinase Couples Actin and Adhesion Dynamics to Survival Signaling,” Molecular and Cellular Biology, Vol. 24, No. 18, 2004, pp. 8113-8133. doi:10.1128/MCB.24.18.8113-8133.2004
[11] B. van de Water, F. Houtepen, M. Huigsloot and I. B. Tijdens, “Suppression of Chemically Induced Apoptosis but Not Necrosis of Renal Proximal Tubular Epithelial (LLC-PK1) Cells by Focal Adhesion Kinase (FAK). Role of FAK in Maintaining Focal Adhesion Organization after Acuterenal Cell Injury,” The Journal of Biological Chemistry, Vol. 276, No. 39, 2001, pp. 36183-36193. doi:10.1074/jbc.M102091200
[12] D. Ilic, E. A. Almeida, D. D. Schlaepfer, P. Dazin, S. Aizawa and C. H. Damsky, “Extracellular Matrix Survival Signals Transduced by Focal Adhesion Kinase Suppress p53-Mediated Apoptosis,” The Journal of Cell Biology, Vol. 143 , No. 2, 1998, pp. 547-560. doi:10.1083/jcb.143.2.547
[13] S. M. Frisch, K. Vuori, E. Ruoslahti and P. Y. Chan-Hui, “Control of Adhesion-Dependent Cell Survival by Focal Adhesion Kinase,” The Journal of Cell Biology, Vol. 134, No. 3, 1996, pp. 793-799. doi:10.1083/jcb.134.3.793
[14] J. E. Hungerford, M. T. Compton, M. L. Matter, B. G. Hoffstrom and C. A. Otey, “Inhibition of pp125FAK in Cultured Fibroblasts Results in Apoptosis,” The Journal of Cell Biology, Vol. 135 , No. 5, 1996, pp. 1383-1390. doi:10.1083/jcb.135.5.1383
[15] L. H. Xu, X. Yang, C. A. Bradham, D. A. Brenner, A. S. Baldwin Jr., R. J. Craven and W. G. Cance, “The Focal Adhesion Kinase Suppresses Transformation-Associated, Anchorage-Independent Apoptosis in Human Breast Cancer Cells. Involvement of Death Receptor-Related Signaling Pathways,” The Journal of Biological Chemistry, Vol. 275, No. 39, 2000, pp. 30597-30604. doi:10.1074/jbc.M910027199
[16] M. S. Duxbury, H. Ito, M. J. Zinner, S. W. Ashley and E. E. Whang, “Focal Adhesion Kinase Gene Silencing Promotes Anoikis and Suppresses Metastasis of Human Pancreatic Adenocarcinoma Cells,” Surgery, Vol. 135, No. 5, 2004, pp. 555-562. doi:10.1016/j.surg.2003.10.017
[17] R. Braren, H. Hu, Y. H. Kim, H. E. Beggs, L. F. Reichardt and R. Wang, “Endothelial FAK Is Essential for Vascular Network Stability, Cell Survival, and Lamellipodial Formation,” The Journal of Cell Biology, Vol. 172 , No. 1, 2006, pp. 151-162. doi:10.1083/jcb.200506184
[18] D. Wang, J. R. Grammer, C. S. Cobbs, J. E. Stewart Jr., Z. Liu, R. Rhoden, T. P. Hecker, Q. Ding and C. L. Gladson, “p125 Focal Adhesion Kinase Promotes Malignant Astrocytoma Cell Proliferation in Vivo,” Journal of Cell Science, Vol. 113, No. 23, 2000, pp. 4221-4230.
[19] J. H. Zhao, H. Reiske and J. L. Guan, “Regulation of the Cell Cycle by Focal Adhesion Kinase,” The Journal of Cell Biology, Vol. 143, No. 7, 1998, pp. 1997-2008. doi:10.1083/jcb.143.7.1997
[20] M. Oktay, K. K. Wary, M. Dans, R. B. Birge and F. G. Giancotti, “Integrinmediated Activation of Focal Adhesion Kinase Is Required for Signaling to Jun NH2-Terminal Kinase and Progression through the G1 Phase of the Cell Cycle,” The Journal of Cell Biology, Vol. 145, No. 7, 1999, pp. 1461-1469. doi:10.1083/jcb.145.7.1461
[21] J. Zhao, R. Pestell and J. L. Guan, “Transcriptional Activation of Cyclin D1 Promoter by FAK Contributes to Cell Cycle Progression,” Molecular Biology of the Cell, Vol. 12, No. 12, 2001, pp. 4066-4077.
[22] M. Toutant, A. Costa, J. M. Studler, G. Kadare, M. Carnaud and J. A. Girault, “Alternative Splicing Controls the Mechanisms of FAK Autophosphorylation,” Molecular and Cell Biology, Vol. 22, No. 22, 2002, pp. 7731-7743. doi:10.1128/MCB.22.22.7731-7743.2002
[23] E. Liu, J. F. Cote and K.Vuori, “Negative Regulation of FAK Signaling by SOCS Proteins,” EMBO Journal, Vol. 22, No. 19, 2003, pp. 5036-5046. doi:10.1093/emboj/cdg503
[24] N. O. Carragher, M. A. Westhoff, V. J. Fincham, M. D. Schaller and M. C. Frame, “A Novel Role for FAK as a Protease Targeting Adaptor Protein. Regulation by p42 ERK and Src,” Current Biology, Vol. 13, No. 16, 2003, pp.1442-1450. doi:10.1016/S0960-9822(03)00544-X
[25] A. Katsumi, A. W. Orr, E.Tzima and M. A. Schwartz, “Integrins in Mechanotransduction,” The Journal of Biological Chemistry, Vol. 279, No. 13, 2004, pp. 12001-12004. doi:10.1074/jbc.R300038200
[26] L. V. Owens, L. Xu, R. J. Craven, G. A. Dent, T. M. Weiner, L. Kornberg, E. T. Liu and W. G. Cance, “Overexpression of the Focal Adhesion Kinase (p125FAK) in Invasive Human Tumors,” Cancer Research, Vol. 55, No. 13, 1995, pp. 2752-2755.
[27] X. J. Liu, L. Yang, H. B. Wu, O. Qiang, M. H. Huang and Y. P. Wang, “Apoptosis of Rat Hepatic Stellate Cells Induced by Anti-Focal Adhesion Kinase Antibody,” World Journal of Gastroenterology, Vol. 8, No. 1, 2002, pp. 734-738.
[28] L. H. Xu, L. V. Owens, G. C. Sturge, X. Yang, E. T. Liu, R. J. Craven and W. G. Cance, “Attenuation of the Expression of the Focal Adhesion Kinase Induces Apoptosis in Tumor Cells,” Cell Growth and Differentiation, Vol. 7, No. 4, 1996, pp. 413-418.
[29] G. Jones, J. Machado Jr., M. Tolnay and A. Merlo, “PTEN-Independent Induction of Caspase-Mediated Cell Death and Reduced Invasion by the Focal Adhesion Targeting Domain (FAT) in Human Astrocytic Brain Tumors Which Highly Express Focal Adhesion Kinase (FAK),” Cancer Research, Vol. 61, No. 15, 2001, pp. 5688-5691.
[30] S. M. Frisch, K. Vuori, E. Ruoslahti and P. Y. Chan-Hui, “Control of Adhesion-Dependent Cell Survival by Focal Adhesion Kinase,” The Journal of Cell Biology, Vol. 134, No. 3, 1996, pp. 793-799. doi:10.1083/jcb.134.3.793
[31] J. E. Hungerford, M. T. Compton, M. L. Matter, B. G. Hoffstrom and C. A. Otey, “Inhibition of pp125FAK in Cultured Fibroblasts Results in Apoptosis,” The Journal of Cell Biology, Vol. 135, No. 5, 1996, pp. 1383-1390. doi:10.1083/jcb.135.5.1383
[32] L. H. Xu, X. Yang, C. A. Bradham, D. A. Brenner, A. S. Baldwin Jr., R. J. Craven and W. G. Cance, “The Focal Adhesion Kinase Suppresses Transformation-Associated, Anchorage-Independent Apoptosis in Human Breast Cancer Cells. Involvement of Death Receptor-Related Signaling Pathways,” The Journal of Biological Chemistry, Vol. 275, No. 39, 2000, pp. 30597-30604. doi:10.1074/jbc.M910027199
[33] M. S. Duxbury, H. Ito, M. J. Zinner, S. W. Ashley and E. E. Whang, “Focal adhesion Kinase Gene Silencing Promotes Anoikis and Suppresses Metastasis of Human Pancreatic Adenocarcinoma Cells,” Surgery, Vol. 135, No. 5, 2004, pp. 555-562. doi:10.1016/j.surg.2003.10.017
[34] M. J. van Nimwegen, S. Verkoeijen, L. van Buren, D. Burg and B. van de Water, “Requirement for Focal Adhesion Kinase in the Early Phase of Mammary Adenocarcinoma Lung Metastasis Formation,” Cancer Research, Vol. 65, No. 11, 2005, pp. 4698-4706. doi:10.1158/0008-5472.CAN-04-4126
[35] S. K. Mitra, S. T. Lim, A. Chi and D. D. Schlaepfer, “Intrinsic Focal Adhesion Kinase Activity Controls Orthotopic Breast Carcinoma Metastasis via the Regulation of Urokinase Plasminogen Activator Expression in a Syngeneic Tumor Model,” Oncogene, Vol. 25, No. 32, 2006, pp. 4429-4440. doi:10.1038/sj.onc.1209482
[36] T. M.Weiner, E. T. Liu, R. J. Craven and W. G. Cance, “Expression of Growth Factor Receptors, the Focal Adhesion Kinase, and Other Tyrosine Kinases in Human Soft Tissue Tumors,” Annuls of Surgical Oncology, Vol. 1, No. 1, 1994, pp.18-27. doi:10.1007/BF02303537
[37] M. Agochiya, V. G. Brunton, D. W.Owens, E. K. Parkinson, C. Paraskeva, W. N. Keith and M. C. Frame, “Increased Dosage and Amplification of the Focal Adhesion Kinase Gene in Human Cancer Cells,” Oncogene, Vol. 18, No. 41, 1999, pp. 5646-5653. doi:10.1038/sj.onc.1202957
[38] Y. Pylayeva, K. M. Gillen, W. Gerald, H. E. Beggs, L. F. Reichardt and F. G. Giancotti, “Ras- and PI3K-Dependent Breast Tumorigenesis in Mice and Humans Requires Focal Adhesion Kinase Signaling,” The Journal of Clinical Investigation, Vol. 119 , No. 2, 2009, pp. 252-266.
[39] S. E. Theocharis, J. T. Klijanienko, E. Padoy, S. Athanassiou, X. X. Sastre-Garau, “Focal Adhesion Kinase (FAK) Immunocytochemical Expression in Breast Ductal Invasive Carcinoma (DIC): Correlation with Clinicopathological Parameters and Tumor Proliferative Capacity,” Medicine Science Monitor, Vol. 15, No. 8, 2009, pp. 221-226.
[40] G. W. Cance, J. E. Harris, M. V. Iacocca, E. Roche, X. H. Yang, J. Chang, S. Simkins and L. X, “Immunohistochemical Analyses of Focal Adhesion Kinase Expression in Benign and Malignant Human Breast and Colon Tissues: Correlation with Preinvasive and Invasive Phenotypes,” Clinical Cancer Research, Vol. 6, 2000, pp. 2417-2423.
[41] M. Ayaki, K. Komatsu, M. Mukai, K. Murata, M. Kameyama, S. Ishiguro, J. Miyoshi, M. Tatsuta and H. Nakamura, “Reduced Expression of Focal Adhesion Kinase in Liver Metastases Compared with Matched Primary Human Colorectal Adenocarcinomas,” Clinical Cancer Research, Vol. 7, No. 10, 2001, pp. 3106-3112.
[42] K. J. Schmitz, F. Grabellus, R. Callies, F. Otterbach, J. Wohlschlaeger, B. Levkau, R. Kimmig, K. W. Schmid and H. A. Baba, “High Expression of Focal Adhesion Kinase (p125FAK) in Node-Negative Breast Cancer Is Related to Overexpression of HER-2/neu and Activated Akt Kinase but Does Not Predict Outcome,” Breast Cancer Res, Vol. 7, No. 2, 2005, pp. 194-203. doi:10.1186/bcr977
[43] A. L. Lark, C. A. Livasy, L. Dressler, D. T. Moore, R. C. Millikan, J. Geradts, M. Iacocca, D. Cowan, D. Little, R. J. Craven and W. Cance, “High Focal Adhesion Kinase Expression in Invasive Breast Carcinomas Is Associated with an Aggressive Phenotype,” Modern Pathology, Vol. 18, No. 10, 2005, pp. 1289-1294. doi:10.1038/modpathol.3800424
[44] S. Ménard, P. Casalini, M. Campiglio, S. M. Pupa and E. Tagliabue, “Role of HER2/neu in Tumor Progression and Therapy,” Cellular and Molecular Life Science, Vol. 61, No. 23, 2004, pp. 2965-2978.
[45] B. J. Deroo and K. S. Korach, “Estrogen Receptors and Human Disease,” Journal of Clinical Investment, Vol. 116, No. 3, 2006, pp. 561-567. doi:10.1172/JCI27987
[46] W. L. McGuire and G. M. Clark, “The Prognostic Role of Progesterone Receptors in Human Breast Cancer,” Seminars in Oncology, Vol. 10, Supplement 4, 1983, pp. 2-6.
[47] V. Thamilselvan, D. H. Craig and M. D. Basson, “FAK Association with Multiple Signal Proteins Mediates Pressure-Induced Colon Cancer Cell Adhesion via a Src-Dependent PI3K/Akt Pathway,” The FASEB Journal, Vol. 21, No. 8, 2007, pp. 1730-1741. doi:10.1096/fj.06-6545com
[48] K. Maung, D. J. Easty, S. P. Hill and D. C. Bennett, “Requirement for Focal Adhesion Kinase in Tumor Cell Adhesion,” Oncogene, Vol. 18, No. 48, 1999, pp. 6824-6828. doi:10.1038/sj.onc.1203094
[49] D. D. Schlaepfer, S. K. Mitra and D. Ilic, “Control of Motile and Invasive Cell Phenotypes by Focal Adhesion Kinase,” Biochimica et Biophysica Acta, Vol. 1692, No. 2-3, 2004, pp. 77-102. doi:10.1016/j.bbamcr.2004.04.008
[50] C. R. Hauck, D. A. Hsia, X. S. Puente, D. A. Cheresh and D. D. Schlaepfer, “FRNK blocks v-Src-Stimulated Invasion and Experimental Metastases without Effects on cell Motility or Growth,” The EMBO Journal, Vol. 21, No. 23, 2002, pp. 6289-6302. doi:10.1093/emboj/cdf631
[51] T. Nagy, H. Wei, T. L. Shen, X. Peng, C. C. Liang, B. Gan and J. L. Guan, “Mammary Epithelial-Specific Deletion of the Focal Adhesion Kinase Gene Leads to Severe Lobulo-Alveolar Hypoplasia and Secretory Immaturity of the Murine Mammary Gland,” The Journal of Biological Chemistry, Vol. 282 , No. 43, 2007, pp. 31766-31776. doi:10.1074/jbc.M705403200
[52] V. M. Golubovskaya, F. A. Kweh and W. G. Cance, “Focal Adhesion Kinase and Cancer,” Histology and Histopathology, Vol. 24, No. 4-6, 2009, pp. 503-510.
[53] J. Zhao and J. L. Guan, “Signal Transduction by Focal Adhesion Kinase in Cancer,” Cancer Metastasis Reviews, Vol. 28, No. 1-2, 2009, pp. 35-49. doi:10.1007/s10555-008-9165-4
[54] D. O. Watermann, B. Gabriel, M. Jager, M. Orlowska-Volk, A. Hasenburg, A. zur Hausen, G. Gitsch and E. Stickeler, “Specific Induction of pp125 Focal Adhesion Kinase in Human Breast Cancer,” British Journal of Cancer, Vol. 93 , No. 6, 2005, pp. 694-698. doi:10.1038/sj.bjc.6602744
[55] M. H. Oktay, K. Oktay, D. Hamele-Bena, A. Buyuk and L.G. Koss, “Focal Adhesion Kinase as a Marker of Malignant Phenotype in Breast and Cervical Carcinomas,” Human Pathology, Vol. 34, No. 3, 2003, pp. 240-245. doi:10.1053/hupa.2003.40
[56] B. Gabriel, A. zur Hausen, E. Stickeler, C. Dietz, G. Gitsch, D. C. Fischer, J. Bouda, C. Tempfer and A. Hasenburg, “Weak Expression of Focal Adhesion Kinase (pp125FAK) in Patients with Cervical Cancer Is Associated with Poor Disease Outcome,” Clinical Cancer Research, Vol. 12, No. , 2006, pp. 2476-2483. doi:10.1158/1078-0432.CCR-05-1867
[57] S. K. Mitra and D. D. Schlaepfer, “Integrin-Regulated FAK-Src Signaling in Normal and Cancer Cells,” Current Opinion in Cell Biology. Vol. 18, No. 5, 2006, pp. 516-523. doi:10.1016/j.ceb.2006.08.011
[58] H. R. Reiske, S. C. Kao, L. A. Cary, J. L. Guan, J. F. Lai and H. C. Chen, “Requirement of Phosphatidylinositol 3-Kinase in Focal Adhesion Kinase-Promoted Cell Migration,” The Journal of Biological Chemistry, Vol. 274, No. 18, 1999, pp. 12361-12366. doi:10.1074/jbc.274.18.12361
[59] R. S. Sawhney, M. M. Cookson, Y. Omar, J. Hauser and M. G. Brattain, “Integrin Alpha2-Mediated ERK and Calpain Activation Play a Critical Role in Cell Adhesion and Motility via Focal Adhesion Kinase Signaling: Identification of a Novel Signaling Pathway,” The Journal of Biological Chemistry, Vol. 281, No. 13, 2006, pp. 8497-8510. doi:10.1074/jbc.M600787200
[60] L. H. Xu, X.Yang, C. A. Bradham, D. A. Brenner, A. S. Baldwin Jr., R. J. Craven and W. G. Cance, “The Focal Adhesion Kinase Suppresses Transformation-Associated, Anchorage-Independent Apoptosis in Human Breast Cancer Cells. Involvement of Death Receptor-Related Signaling Pathways,” The Journal of Biological Chemistry, Vol. 275, No. 39, 2000, pp. 30597-30604. doi:10.1074/jbc.M910027199
[61] N. Benlimame, Q. He, S. Jie, D. Xiao, Y. J. Xu, M. Loignon, D. D. Schlaepfer and M. A. Alaoui-Jamali, “FAK Signaling Is Critical for ErbB-2/ErbB-3 Receptor Cooperation for Oncogenic Transformation and Invasion,” The Journal of Cell Biology, Vol. 171, No. 3, 2005, pp. 505-516. doi:10.1083/jcb.200504124
[62] Y. Xu, N. Benlimame, J. Su, Q. He and M. A. Alaoui-Jamali, “Regulation of Focal Adhesion Turnover by ErbB Signalling in Invasive Breast Cancer Cells,” British Journal of Cancer, Vol. 100, No. 4, 2009, pp. 633-643. doi:10.1038/sj.bjc.6604901
[63] M. D. Planas-Silva, R. D. Bruggeman, R. T. Grenko and J. Stanley Smith, “Role of c-Src and Focal Adhesion Kinase in Progression and Metastasis of Estrogen Receptor-Positive Breast Cancer,” Biochemical and Biophysical Research Communications, Vol. 341, No. 1, 2006, pp. 73-81. doi:10.1016/j.bbrc.2005.12.164
[64] V. C. Lin, E. H. Ng, S. E. Aw, M. G. Tan, E. H. Ng and B. H. Bay, “Progesterone Induces Focal Adhesion in Breast Cancer Cells MDA-MB-231 Transfected with Progesterone Receptor Complementary DNA,” Molecular Endocrinology, Vol. 14, No. 3, 2000, pp. 348-358. doi:10.1210/me.14.3.348

  
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