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. 2017 Mar 21;7(5):627-635.
doi: 10.1002/2211-5463.12122. eCollection 2017 May.

Screening and identification of the tumor-associated antigen CK10, a novel potential liver cancer marker

Zhaoyu Zhu  1   2 , Zheng Liu  1 , Yuchun Liu  1 , Chunyan Wang  1 , Ruyu Li  1 , Hui Liu  1 , Baohua Gu  1 , Guodong Li  3 , Shoutao Zhang  1   4
Affiliations

Screening and identification of the tumor-associated antigen CK10, a novel potential liver cancer marker

Zhaoyu Zhu et al. FEBS Open Bio. .

Abstract

Hepatocellular carcinoma (HCC) is a malignancy that is associated with high mortality rates in Asia. These tumors are highly invasive and their etiology is frequently unknown. Thus, most patients are diagnosed in the middle and late stages of the disease, and thus do not have sufficient time for therapy. Therefore, it is essential to study the early diagnosis and treatment of HCC; in this regard, the study of tumor-associated antigens has received much attention. Here, antigens from the human primary HCC cell line, QGY-7703, were used to immunize mice in order to prepare monoclonal antibodies. The specific antigen recognized by antibody 11C3 was purified from total protein lysates of QGY-7703 by immunoaffinity chromatography. The validity of the candidate antigen as a new HCC-associated marker was tested using SDS/PAGE, western blot, HPLC-ESI-MS/MS, and RT-qPCR. Our results showed that the levels of CK10 in HCC-derived cell lines were significantly higher than those in normal liver cells. Thus, we suggest that CK10 may be involved in the formation and development of HCC, and may be a therapeutically targetable tumor-associated antigen.

Keywords: antibody; biomarker; hepatocellular carcinoma; tumor‐associated antigen.

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Figures

Figure 1
Figure 1
SDS/PAGE of monoclonal antibody 11C3 purified by Protein A.
Figure 2
Figure 2
SDS/PAGE and western blotting analysis of total cellular protein from QGY‐7703 human hepatocellular cancer cells and L02 human normal liver cells. Lane M, marker; Lane 1, total cellular protein of QGY‐7703 cells; Lane 2, total cellular protein of L02 cells; Lane 3, western blotting of QGY‐7703 total cellular protein; Lane 4, western blotting of L02 total cellular protein.
Figure 3
Figure 3
SDS/PAGE and western blotting analysis of the purified antigen of 11C3. Lane M, marker; Lane 1, concentration of elution peak by immunoaffinity chromatography; Lane 2, western blotting of antigen 11C3 purified by immunoaffinity chromatography.
Figure 4
Figure 4
Western blotting analysis of the antigen:antibody reaction between the purified antigen and the commercial anti‐CK10 monoclonal antibody. Lane 1, CK10; Lane 2, CK8.
Figure 5
Figure 5
The analysis of relative expression level changes of CK10 mRNA in QGY‐7703 and L02. The GAPDH was used as the endogenous reference in three groups of parallel control experiments.
See this image and copyright information in PMC

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