J Korean Med Sci.  2012 Dec;27(12):1479-1485. 10.3346/jkms.2012.27.12.1479.

Candidates for Tumor Markers of Cervical Cancer Discovered by Proteomic Analysis

• Song JY
• Bae HS
• Koo DH
• Lee JK
• Jung HH
• Lee KW
• Lee NW

• KMID: 2157970
• DOI: http://doi.org/10.3346/jkms.2012.27.12.1479

Abstract

Cervical cancer is the second most common gynecological cancer among Korean women. While nationwide screening program has developed, the pathogenesis of cervical cancer is unknown. The aim of this study was to compare the protein expression profiles between cervical squamous carcinomas and normal cervical tissues in order to identify proteins that are related to the cancer. Three cervical cancer tissue samples and three normal cervical tissue samples were obtained and protein expression was compared and was identified in the samples with the use of matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). A total of 20 proteins that showed up-regulated expression in the cervical cancer tissue samples were selected and identified. Seven proteins were matched to allograft inflammatory factor 1 (AIF-1), actine-like protein 2 (ALP2), brain type fatty acid-binding protein (B-FABP), NCK adaptor protein 1 (NCK-1), islet cell autoantigen 1 (ICA69), cationic trypsinogen (PRSS1), and cyclin-dependent kinase 4 (CDK4), but the remaining 13 proteins were unidentifiable. After confirmation by RT-PCR, Western blotting and immunohistochemistry, we found that B-FABP, NCK-1, and CDK4 were related to the pathogenesis of cervical cancer. These proteins are suggested as candidates of new pathological tumor markers for cervical cancer.

Keyword

Cervical Cancer; Two-Dimensional Polyacrylamide Gel Electrophoresis; Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry

MeSH Terms

Adaptor Proteins, Signal Transducing/genetics/metabolism
Blotting, Western
Carcinoma, Squamous Cell/*metabolism/pathology
Cyclin-Dependent Kinase 4/genetics/metabolism
Electrophoresis, Gel, Two-Dimensional
Fatty Acid-Binding Proteins/genetics/metabolism
Female
Humans
Immunohistochemistry
Oncogene Proteins/genetics/metabolism
*Proteomics
Reverse Transcriptase Polymerase Chain Reaction
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Tumor Markers, Biological/genetics/*metabolism
Uterine Cervical Neoplasms/*metabolism/pathology
Adaptor Proteins, Signal Transducing
Fatty Acid-Binding Proteins
Oncogene Proteins
Tumor Markers, Biological
Cyclin-Dependent Kinase 4

Figure

• 

  Fig. 1 Comparison of 2D PAGE proteins separated from cervical cancer tissue and normal cervical tissue samples. A total of 20 over-expressed protein spots in cervical cancer tissues were analyzed by MALDI-TOF-MS. Of the 20 proteins, seven proteins were matched to AIF-1, ALP2, B-FABP, NCK-1, ICA69, PRSS1, and CDK4 and 13 of these proteins could not be matched, even after PCR amplification of possible genes. MW, molecular weight; AIF-1, allograft inflammatory factor 1; ALP2, actin-like protein 2; B-FABP, brain type fatty acid binding protein; NCK-1, NCK adaptor protein 1; ICA69, islet cell autoantigen; PRSS1, cationic trypsinogen; CDK4, cyclin-dependent kinase 4.

• 

  Fig. 2 RT-PCR demonstrates that the expression levels of AIF-1, ALP2, B-FABP, NCK-1, ICA69, PRSS1, and CDK4 are higher in cervical cancer tissues than in normal cervical tissues. β2-microglobulin is evenly expressed in both normal and cervical cancer tissues. AIF-1, allograft inflammatory factor 1; ALP2, actin-like protein 2; B-FABP, brain type fatty acid binding protein; NCK-1, NCK adaptor protein 1; ICA69, islet cell autoantigen; PRSS1, cationic trypsinogen; CDK4, cyclin-dependent kinase 4; L, ladder (100 bp); cervical cancers (C1, C2); normal cervical tissues (N1, N2).

• 

  Fig. 3 Western blotting shows distinct protein bands on autoradiographs corresponding to NCK-1, B-FABP, and CDK-4, which are observed at approximately 15 kDa, 15 kDa, and 15 kDa in cervical squamous cell carcinoma (C1, C2), but the expression levels are low in normal cervical tissues (N1, N2). NCK-1, NCK adaptor protein 1; B-FABP, brain type fatty acid binding protein; CDK4, cyclin-dependent kinase 4.

• 

  Fig. 4 Immunostaining of NCK-1, B-FABP, and CDK4 are not observed in normal cervical tissues, but positive staining is observed in cervical cancer tissues. (A, C) and (E) normal cervical tissues with negative staining of NCK-1, B-FABP, and CDK. (B, D) and (F) cervical cancer tissues with positive staining of NCK-1, B-FABP, and CDK. B-FABP, brain type fatty acid binding protein; NCK-1, NCK adaptor protein 1; CDK4, cyclin-dependent kinase 4.

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