Abstract
Tumor-associated antigens (TAAs) can be used as cancer markers and as signposts of therapeutic targets since their inimitable expression in cancer or significant overexpression in esophageal squamous cell carcinoma (ESCC) correlates with the initiation and progression of the diseases. Immunoblotting, also known as Western blotting or protein blotting, is a core technique in cell and molecular biology to detect proteins and glycoproteins. The technique allows detection of TAAs from complex protein samples such as in serum, aspirate, or solid tumor homogenate. In the process, proteins are separated according to the molecular weight. They were visualized within a gel matrix and then transferred to a supporting membrane. Finally, they are probed for binding with corresponding antibodies and identified the target proteins. Herein, we describe the Western blots analysis to detect protein or glycoprotein in samples from patients with esophageal squamous cell carcinoma (ESCC) or cells derived from ESCC.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Harao M, Mittendorf EA, Radvanyi LG (2015) Peptide-based vaccination and induction of CD8+ T-cell responses against tumor antigens in breast cancer. BioDrugs 29:15–30
Gao HJ, Yue ZG, Zheng M, Zheng ZX (2012) Identification and expression of a tumor-associated antigen in esophageal squamous cell carcinoma. Zhongguo Yi Xue Ke Xue Yuan Xue Bao 34:244–248
Lee SY, Jeoung D (2007) The reverse proteomics for identification of tumor antigens. J Microbiol Biotechnol 17:879–890
Fernandez MF, Tang N, Alansari H, Karvonen RL, Tomkiel JE (2005) Improved approach to identify cancer-associated autoantigens. Autoimmun Rev 4:230–235
Liu CC, Yang H, Zhang R, Zhao JJ, Hao DJ (2017) Tumour-associated antigens and their anti-cancer applications. Eur J Cancer Care 26:e12446
Zhang JY, Looi KS, Tan EM (2009) Identification of tumor-associated antigens as diagnostic and predictive biomarkers in cancer. Methods Mol Biol 520:1–10
Xu WW, Li B, Lam AK, Tsao SW, Law SY, Chan KW, Yuan QJ, Cheung AL (2015) Targeting VEGFR1- and VEGFR2-expressing non-tumor cells is essential for esophageal cancer therapy. Oncotarget 6:1790–1805
Islam F, Gopalan V, Law S, Tang JC, Lam AK (2019) FAM134B promotes esophageal squamous cell carcinoma in vitro and its correlations with clinicopathologic features. Hum Pathol 87:1–10
Yu VZ, Wong VC, Dai W, Ko JM, Lam AK, Chan KW, Samant RS, Lung HL, Shuen WH, Law S, Chan YP, Lee NP, Tong DK, Law TT, Lee VH, Lung ML (2015) Nuclear localization of DNAJB6 is associated with survival of patients with esophageal cancer and reduces akt signaling and proliferation of cancer cells. Gastroenterology 149:1825–1836.e5
Chai AW, Cheung AK, Dai W, Ko JM, Ip JC, Chan KW, Kwong DL, Ng WT, Lee AW, Ngan RK, Yau CC, Tung SY, Lee VH, Lam AK, Pillai S, Law S, Lung ML (2016) Metastasis-suppressing NID2, an epigenetically-silenced gene, in the pathogenesis of nasopharyngeal carcinoma and esophageal squamous cell carcinoma. Oncotarget 7:78859–78871
Li B, Xu WW, Lam AKY, Wang Y, Hu HF, Guan XY, Qin YR, Saremi N, Tsao SW, He QY, Cheung ALM (2017) Significance of PI3K/AKT signaling pathway in metastasis of esophageal squamous cell carcinoma and its potential as a target for anti-metastasis therapy. Oncotarget 8:38755–38766
Gopalan V, Islam F, Pillai S, Tang JC, Tong DK, Law S, Chan KW, Lam AK (2016) Overexpression of microRNA-1288 in oesophageal squamous cell carcinoma. Exp Cell Res 348:146–154
Islam F, Gopalan V, Law S, Tang JC, Chan KW, Lam AK (2017) MiR-498 in esophageal squamous cell carcinoma: clinicopathological impacts and functional interactions. Hum Pathol 62:141–151
Ng HY, Li J, Tao L, Lam AK, Chan KW, Ko JMY, Yu VZ, Wong M, Li B, Lung ML (2018) Chemotherapeutic treatments increase PD-L1 expression in esophageal squamous cell carcinoma through EGFR/ERK activation. Transl Oncol 11:1323–1333
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Islam, F., Gopalan, V., Lam, A.K. (2020). Immunoblotting in Detection of Tumor-Associated Antigens in Esophageal Squamous Cell Carcinoma. In: Lam, A. (eds) Esophageal Squamous Cell Carcinoma. Methods in Molecular Biology, vol 2129. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0377-2_20
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0377-2_20
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0376-5
Online ISBN: 978-1-0716-0377-2
eBook Packages: Springer Protocols