Microquantitation of Van Gogh-like Protein 1 by Using Antibody-Conjugated Magnetic Beads

다국어 초록 (Multilingual Abstract)

Van Gogh-like protein 1 (VANGL1) is an integral membrane protein that has a decisive effect on invasion, migration, and metastasis of tumor cells in various cancers including colorectal cancer. Elevation of VANGL1 level in cancer tissue is often observed with progressive and metastatic colorectal cancer. Therefore, detection of VANGL1 level in patient specimens can be used as a diagnostic marker for colorectal cancer screening. In this study, we developed magnetic beads that conjugate the VANGL1 antibody, which can be used for quantitative analysis of VANGL1. The procedure for bead preparation was optimized, and detection analysis was validated by using Caco2 and HT29 colorectal cancer cell lysates. Through the procedure, VANGL1 level from cell lysates of Caco2 and HT29 cells were quantified to estimate whether the antibody-conjugated magnetic bead can be used to trace amounts of VANGL1. Results from VANGL1 level obtained by using the antibody-conjugated magnetic beads suggest that the procedure has high precision and sensitivity in analyzing VANGL1. In conclusion, the results indicate that the method is appropriate for microquantitation of VANGL1 from patient specimens.

참고문헌 (Reference)

1 Wang, Y., "Wnt/Planar cell polarity signaling: A new paradigm for cancer therapy" 8 : 2103-2109, 2009

2 Kim, H., "Presenilin-1D257A and D385A mutants fail to cleave Notch in their endoproteolyzed forms, but only presenilin-1D385A mutant can restore its gamma-secretase activity with the compensatory overexpression of normal C-terminal fragment" 280 : 22462-22472, 2018

3 Lee, S-D., "Microquantitation of Prostate-Specific Antigen by Using Antibody-Conjugated Magnetic Microsphere Beads" 18 : 1474-1477, 2018

4 Kho, D. H., "KITENIN recruits Dishevelled/PKCd to form a functional complex and controls the migration and invasiveness of colorectal cancer cells" 58 : 509-519, 2009

5 Lee, K. H., "KITENIN promotes glioma invasiveness and progression, associated with the induction of EMT and stemness markers" 6 : 3240-3253, 2015

6 Ryu, H. S., "KITENIN is associated with tumor progression in human gastric cancer" 30 : 3479-3486, 2010

7 Cho, S. B., "KITENIN is associated with activation of AP-1target genes via MAPK cascades signaling in human hepatocellular carcinoma progression" 19 : 115-123, 2011

8 Lee, J. K., "KITENIN increases invasion and migration of mouse squamous cancer cells and promotes pulmonary metastasis in a mouse squamous tumor model" 583 : 711-717, 2009

9 Sun, E. G., "KITENIN functions as a fine regulator of ErbB4 expression level in colorectal cancer via protection of ErbB4 from E3-ligase Nrdp1-mediated degradation" 56 : 1068-1081, 2017

10 Lee, J. H., "KAI1 COOHterminal interacting tetraspanin (KITENIN), a member of the tetraspanin family, interacts with KAI1, a tumor metastasis suppressor, and enhances metastasis of cancer" 64 : 4235-4243, 2004

11 Lee, J.K., "KAI1 COOH-terminal interacting tetraspanin (KITENIN) expression in early and advanced laryngeal cancer" 120 : 2010

12 Yagyu, R., "Isolation and characterization of a novel human gene, VANGL1, as a therapeutic target for hepatocellular carcinoma" 20 : 1173-1178, 2002

13 Oh, H. H., "Impact of KITENIN on tumor angiogenesis and lymphangiogenesis in colorectal cancer" 35 : 253-260, 2016

14 Rowe, A., "Expression of KITENIN, a KAI1/CD82 binding protein and metastasis enhancer, in bladder cancer cell lines: Relationship to KAI1/CD82 levels and invasive behaviour" 16 : 1267-1272, 2006

15 Lee, S., "Expression of KITENIN in human colorectal cancer and its relation to tumor behavior and progression" 6 : 210-220, 2011

16 Lee, S., "Expression of KITENIN in human colorectal cancer and its relation to tumor behavior and progression" 61 : 210-220, 2011

17 Yoon, T. M., "Expression of KITENIN and its association with tumor progression in oral squamous cell carcinoma" 40 : 222-226, 2013

18 Cetin, G. O., "Downregulation of VANGL1 Inhibits Cellular Invasion Rather than Cell Motility in Hepatocellular Carcinoma Cells Without Stimulation" 19 : 283-287, 2015

19 Wagner, B., "Development of a multiplex assay for the detection of antibodies to Borrelia burgdorferi in horses and its validation using Bayesian and conventional statistical methods" 144 : 374-381, 2011

20 Bae, J. A., "Biology of Human Tumors Elevated Coexpression of KITENIN and the ErbB4 CYT-2 Isoform Promotes the Transition from Colon Adenoma to Carcinoma Following APC loss" 22 (22): 1284-1294, 2016

21 Houser, B., "Bio-rad’s Bio-Plex® suspension array system, xMAP technology overview" 118 : 192-196, 2012

22 Bae, J. A., "An unconventional KITENIN/ErbB4-mediated downstream signal of EGF upregulates c-Jun and the invasiveness of colorectal cancer cells" 20 : 4115-4128, 2014

23 Anastas, J. N., "A protein complex of SCRIB, NOS1AP and VANGL1 regulates cell polarity and migration and is associated with breast cancer progression" 31 : 3696-3708, 2012

24 Wagner, B., "A fluorescent bead-based multiplex assay for the simultaneous detection of antibodies to B. burgdorferi outer surface proteins in canine serum" 140 : 190-198, 2011