Detection of collagen triple helix repeat containing-1 and nuclear factor (erythroid-derived 2)-like 3 in colorectal cancer
Tóm tắt
Collagen Triple Helix Repeat Containing-1 (CTHRC1) and Nuclear factor (erythroid-derived 2)-like 3 (NFE2L3) may be useful biomarker candidates for the diagnosis of colorectal cancer (CRC) since they have shown an increase messenger RNA transcripts (mRNA) expression level in adenomas and colorectal tumours when compared to normal tissues. To evaluate CTHRC1 and NFE2L3 as cancer biomarkers, it was generated and characterised several novel specific polyclonal antibodies (PAb), monoclonal antibodies (MAbs) and soluble Fab fragments (sFabs) against recombinant CTHRC1 and NFE2L3 proteins, which were obtained from different sources, including a human antibody library and immunised animals. The antibodies and Fab fragments were tested for recognition of native CTHRC1 and NFE2L3 proteins by immunoblotting analysis and enzyme-linked immunosorbent assay (ELISA) in colorectal cell lines derived from tumour and cancer tissues. Both, antibodies and a Fab fragment showed high specificity since they recognised only their corresponding recombinant antigens, but not a panel of different unrelated- and related proteins. In Western blot analysis of CTHRC1, a monoclonal antibody designated CH21D7 was able to detect a band of the apparent molecular weight of a full-length CTHRC1 in the human colon adenocarcinoma cell line HT29. This result was confirmed by a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with the monoclonal antibodies CH21D7 and CH24G2, detecting CTHRC1 in HT29 and in the colon adenocarcinoma cell line SW620. Similar experiments were performed with PAb, MAbs, and sFab against NFE2L3. The immunoblot analysis showed that the monoclonal antibody 41HF8 recognised NFE2L3 in HT29, and leukocytes. These results were verified by DAS-ELISA assay using the pairs PAb/sFab E5 and MAb 41HF8/sFab E5. Furthermore, an immunoassay for simultaneous detection of the two cancer biomarkers was developed using a Dissociation-Enhanced Lanthanide Fluorescent Immunoassay technology (DELFIA). In conclusion, the antibodies obtained in this study are specific for CTHRC1 and NFE2L3 since they do not cross-react with unrelated- and related proteins and are useful for specific measurement of native CTHRC1 and NFE2L3 proteins. The antibodies and immunoassays may be useful for the analysis of CTHRC1 and NFE2L3 in clinical samples and for screening of therapeutic compounds in CRC.
Tài liệu tham khảo
Jemal A, et al: Cancer statistics, 2003. CA Cancer J Clin. 2003, 53 (1): 5-26. 10.3322/canjclin.53.1.5.
Jemal A, et al: Cancer statistics, 2008. CA Cancer J Clin. 2008, 58 (2): 71-96. 10.3322/CA.2007.0010.
Leonard GD, Brenner B, Kemeny NE: Neoadjuvant chemotherapy before liver resection for patients with unresectable liver metastases from colorectal carcinoma. J Clin Oncol. 2005, 23 (9): 2038-2048. 10.1200/JCO.2005.00.349.
Ross JS: Biomarker update for breast, colorectal and non-small cell lung cancer. Drug News Perspect. 2010, 23 (1): 82-10.1358/dnp.2010.23.1.1437316.
Siena S, et al: Biomarkers predicting clinical outcome of epidermal growth factor receptor targeted therapy in metastatic colorectal cancer. J Nat Cancer Inst. 2009, 101 (19): 1308-1324. 10.1093/jnci/djp280.
Walther A, et al: Genetic prognostic and predictive markers in colorectal cancer. Nature Reviews Cancer. 2009, 9 (7): 489-499. 10.1038/nrc2645.
Hammarström S: The carcinoembryonic antigen (CEA) family: structures, suggested functions and expression in normal and malignant tissues. Seminars in Cancer Biol. 1999, 9 (2): 67-81. 10.1006/scbi.1998.0119.
Tanaka T, et al: Biomarkers for colorectal cancer. Int J Mol Sci. 2010, 11 (9): 3209-3225. 10.3390/ijms11093209.
Paik S, et al: A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer. N Engl J Med. 2004, 351 (27): 2817-2826. 10.1056/NEJMoa041588.
Paik S, et al: Gene expression and benefit of chemotherapy in women with node-negative, estrogen receptor-positive breast cancer. J Clin Oncol. 2006, 24 (23): 3726-3734. 10.1200/JCO.2005.04.7985.
Clark-Langone K, et al: Translating tumor biology into personalized treatment planning: analytical performance characteristics of the Oncotype DX(R) Colon Cancer Assay. BMC Cancer. 2010, 10 (1): 691-10.1186/1471-2407-10-691.
Chiu S-T, et al: Clinicopathologic correlation of up-regulated genes identified using cdna microarray and real-time reverse transcription-PCR in human colorectal cancer. Cancer Epidemiol Biomarkers Prev. 2005, 14 (2): 437-443. 10.1158/1055-9965.EPI-04-0396.
Durmus T, et al: Expression analysis of the novel gene collagen triple helix repeat containing-1 (Cthrc1). Gene Expression Patterns. 2006, 6 (8): 935-940. 10.1016/j.modgep.2006.03.008.
Turashvili G, et al: Novel markers for differentiation of lobular and ductal invasive breast carcinomas by laser microdissection and microarray analysis. BMC Cancer. 2007, 7 (55): 1-20.
Ip W, et al: Collagen triple helix repeat containing 1 promotes melanoma cell adhesion and survival. J Cutan Med Surg. 2011, 15 (2): 103-110.
Kobayashi A, et al: Molecular cloning and functional characterization of a New Cap'n'Collar family transcription factor Nrf3. J Biol Chem. 1999, 274 (10): 6443-6452. 10.1074/jbc.274.10.6443.
Sankaranarayanan K, Jaiswal AK: Nrf3 negatively regulates antioxidant-response element-mediated expression and antioxidant induction of NAD(P)H:quinone oxidoreductase1 gene. J Biol Chem. 2004, 279 (49): 50810-50817. 10.1074/jbc.M404984200.
Chenais B, et al: Functional and placental expression analysis of the human NRF3 transcription factor. Mol Endocrinol. 2005, 19 (1): 125-137.
Santin AD: Gene expression profiling in primary ovarian serous papillary tumors and normal ovarian epithelium. 2005, United States Patent US2005004853
Clarke, et al: Compositions and methods for treating and diagnosing cancer. 2007, United States Patent US20070099209
Nakamura, et al: Method for diagnosing testicular seminomas. 2006, United States Patent US20060194199
Chevillard G, Paquet M, Blank V: Nfe2l3 (Nrf3) deficiency predisposes mice to T-cell lymphoblastic lymphoma. Blood. 2011, 117 (6): 2005-2008. 10.1182/blood-2010-02-271460.
Nguewa P, et al: Identification of Importin 8 (IPO8) as the most accurate reference gene for the clinicopathological analysis of lung specimens. BMC Mol Biol. 2008, 9 (1): 103-10.1186/1471-2199-9-103.
Rodriguez M, et al: Epitope mapping of the nucleocapsid protein of European and North American isolates of porcine reproductive and respiratory syndrome virus. J Gen Virol. 1997, 78 (9): 2269-2278.
Sastre P, et al: Differentiation between human coronaviruses NL63 and 229E using a novel double-antibody sandwich enzyme-linked immunosorbent assay based on specific monoclonal antibodies. Clin Vaccine Immunol. 2011, 18 (1): 113-118. 10.1128/CVI.00355-10.
Hoet RM, et al: Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity. Nature Biotechnology. 2005, 23 (3): 344-348. 10.1038/nbt1067.
Nakane PK, Pierce GB: Enzyme-labeled antibodies: preparation and application for the localization of antigens. J Histochem Cytochem. 1966, 14 (12): 929-931. 10.1177/14.12.929.
Bayer EA, Wilchek M: The use of the avidin-biotin complex as a tool in molecular biology. Methods Biochem Anal. 1980, 26: 1-45.
Shin I, et al: Markers for diagnosis of cancer and its use. 2010, European Patent EP2177628A2
Tang L, et al: Aberrant expression of collagen triple helix repeat containing 1 in human solid cancers. Clin Cancer Res. 2006, 12 (12): 3716-3722. 10.1158/1078-0432.CCR-06-0030.
Maes T, et al: Method for colorectal tumor diagnosis. 2008, European Patent EP2008/010665
Price JE, Zhang RD: Studies of human breast cancer metastasis using nude mice. Cancer and Metastasis Reviews. 1990, Netherlands: Springer, 285-297.
Kim J, Stein R, O'Hare M: Three-dimensional in vitro tissue culture models of breast cancer - a review. Breast Cancer Research and Treatment. 2004, Netherlands: Springer, 281-291.
Ertel A, et al: Pathway-specific differences between tumor cell lines and normal and tumor tissue cells. Mol Cancer. 2006, 5 (1): 55-10.1186/1476-4598-5-55.
UniProtKB: Collagen triple helix repeat-containing protein 1 precursor - Homo sapiens. [http://www.uniprot.org/uniprot/Q96CG8]
UniProtKB: Nuclear factor erythroid 2-related factor 3 - Homo sapiens. [http://www.uniprot.org/uniprot/Q9Y4A8]
Zhang Y, et al: The Nrf3 Transcription Factor Is a Membrane-bound Glycoprotein Targeted to the Endoplasmic Reticulum through Its N-terminal Homology Box 1 Sequence. J Biol Chem. 2009, 284 (5): 3195-3210.
Ahmed N, et al: An approach to remove albumin for the proteomic analysis of low abundance biomarkers in human serum. Proteomics. 2003, 3 (10): 1980-1987. 10.1002/pmic.200300465.
The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6890/12/2/prepub