Clin Exp Otorhinolaryngol.  2008 Jun;1(2):97-102. 10.3342/ceo.2008.1.2.97.

Detection of MAGE and SSX Gene Expressions by RT-nested PCR Using Common Primers in Head and Neck Cancer

• Song DW
• Shin SJ
• Kim DE
• Jung SG
• Park JW
• Lee KD

• KMID: 1486061
• DOI: http://doi.org/10.3342/ceo.2008.1.2.97

Abstract

OBJECTIVES
The melanoma antigen gene (MAGE) and synovial sarcoma on X chromosome (SSX) gene families are silent in most normal adult tissues, but are expressed in a variety of malignant lesions. Therefore, detection of MAGE and SSX transcription may be useful for the diagnosis of head and neck cancers. The aim of this study is to detect MAGE and SSX gene transcripts of head and neck cancers using the MAGE 1-6 assay and the SSX 1-9 assay. METHODS: The transcripts of MAGE 1-6 and SSX 1-9 genes were detected by the MAGE 1-6 assay and the SSX 1-9 assay respectively, in cancer cell lines, cancer tissue, and induced sputum specimens from head and neck cancer patients. RESULTS: The transcripts of MAGE 1-6 and SSX 1-9 genes were detected in 82.8% and 75.9% of head and neck cancer tissues (N=29) respectively, and 96.6% of cancer tissues expressed at least one of MAGE 1-6 or SSX 1-9 genes. In the induced sputum of head and neck cancer patients (N=18), the transcripts of MAGE 1-6 and SSX 1-9 genes were detected in 72.2% and 77.8%, respectively, and 94.4% of the sputum specimens were positive for either the MAGE 1-6 or the SSX 1-9 assay. CONCLUSION: These results suggest that the combination of MAGE 1-6 and SSX 1-9 assays may be useful in the diagnosis of head and neck cancer.

Keyword

MAGE and SSX gene; Head and neck cancer; RT-PCR

MeSH Terms

Adult
Cell Line
Gene Expression
Head
Head and Neck Neoplasms
Humans
Melanoma
Neck
Polymerase Chain Reaction
Sarcoma, Synovial
Sputum
X Chromosome

Figure

• 

  Fig. 1 The binding sites of common primers on SSX-1 cDNA (A) and PCR products amplified by MAGE 1-6 and SSX 1-9 assay (B).

• 

  Fig. 2 Detection of SSX 1-9 transcripts of cancer cell lines by SSX 19 assay. M: Marker; 1: SNU 484; 2: SNU 620; 3: SNU 638; 4: SNU 668; 5: SNU C1; 6: SNU C4; 7: SNU C5; 8: HT29; 9: HCT 116; 10: AMC-HN3; 11: AMC-HN4; 12: AMC-HN7; 13: U937; 14: HL 60; 15: Caski; 16: C4-II; 17: ME-180; 18: Hela; 19: CUNC-6; 20: SiHa; 21: NCI-H292; 22; NCI-H522; 23: NCI-H1703; 24: A-549; 25: PC 3; 26: DU 145; 27: HEPG2; 28: SNU 182; 29: SNU 354; 30: SNU 387; 31: SNU 398; 32: SNU 423; 33: HEK 293; 34: MDA 231; 35: SAOS 2.

• 

  Fig. 3 Detection of MAGE 1-6 and SSX 1-9 gene expressions in cancer tissue of the patients with head and neck cancer by MAGE 1-6 assay and SSX 1-9 assay.

• 

  Fig. 4 Detection of the transcripts of MAGE 1-6 and SSX 1-9 genes in sputum containing SNU484 by MAGE 1-6 assay and SSX 1-9 assay.

• 

  Fig. 5 Detection of MAGE 1-6 and SSX 1-9 gene expressions in sputum of the patients with head and neck cancer by MAGE 1-6 assay and SSX 1-9 assay.

• 

  Fig. 6 Representative results of MAGE 1-6 and SSX 1-9 expression in sputum of the patients with benign lung disease (n=22) by MAGE 1-6 or SSX 1-9 assay.

Reference

1. Bodey B. Cancer-testis antigens: promising targets for antigen directed antineoplastic immunotherapy. Expert Opin Biol Ther. 2002; 8. 2(6):577–584. PMID: 12171503.

2. Mashino K, Sadanaga N, Tanaka F, Yamaguchi H, Nagashima H, Inoue H, et al. Expression of multiple cancer-testis antigen genes in gastrointestinal and breast carcinomas. Br J Cancer. 2001; 9. 01. 85(5):713–720. PMID: 11531257.

3. Scanlan MJ, Gure AO, Jungbluth AA, Old LJ, Chen YT. Cancer/testis antigens: an expanding family of targets for cancer immunotherapy. Immunol Rev. 2002; 10. 188:22–32. PMID: 12445278.

4. Tajima K, Obata Y, Tamaki H, Yoshida M, Chen YT, Scanlan MJ, et al. Expression of cancer/testis (CT) antigens in lung cancer. Lung Cancer. 2003; 10. 42(1):23–33. PMID: 14512184.

5. Chen CH, Huang GT, Lee HS, Yang PM, Yan MD, Chen DS, et al. High frequency of expression of MAGE genes in human hepatocellular carcinoma. Liver. 1999; 4. 19(2):110–114. PMID: 10220740.

6. Eura M, Ogi K, Chikamatsu K, Lee KD, Nakano K, Masuyama K, et al. Expression of the MAGE gene family in human head-and-neck squamous-cell carcinomas. Int J Cancer. 1995; 10. 64(5):304–308. PMID: 7591301.

7. Otte M, Zafrakas M, Riethdorf L, Pichlmeier U, Löning T, Jänicke F, et al. MAGE-A gene expression pattern in primary breast cancer. Cancer Res. 2001; 9. 61(18):6682–6687. PMID: 11559535.

8. Park JW, Kwon TK, Kim IH, Sohn SS, Kim YS, Kim CI, et al. A new strategy for the diagnosis of MAGE-expressing cancers. J Immunol Methods. 2002; 8. 266(1-2):79–86. PMID: 12133624.

9. Clark J, Rocques PJ, Crew AJ, Gill S, Shipley J, Chan AM, et al. Identification of novel genes, SYT and SSX, involved in the t(X;18)(p11.2; q11.2) translocation found in human synovial sarcoma. Nat Genet. 1994; 8. 7(4):502–508. PMID: 7951320.

10. Gure AO, Türeci O, Sahin U, Tsang S, Scanlan MJ, Jäger E, et al. SSX: a multigene family with several members transcribed in normal testis and human cancer. Int J Cancer. 1997; 9. 72(6):965–971. PMID: 9378559.

11. Güre AO, Wei IJ, Old LJ, Chen YT. The SSX gene family: characterization of 9 complete genes. Int J Cancer. 2002; 10. 101(5):448–453. PMID: 12216073.

12. Morot-Bizot SC, Talon R, Leroy S. Development of a multiplex PCR for the identification of Staphylococcus genus and four staphylococcal species isolated from food. J Appl Microbiol. 2004; 97(5):1087–1094. PMID: 15479426.

13. Türeci O, Chen YT, Sahin U, Güre AO, Zwick C, Villena C, et al. Expression of SSX genes in human tumors. Int J Cancer. 1998; 7. 77(1):19–23. PMID: 9639388.

14. Matsuda M, Nagumo S, Horai T, Yoshino K. Cytologic diagnosis of laryngeal and hypopharyngeal squamous cell carcinoma in sputum. Acta Cytol. 1988; Sep–Oct. 32(5):655–657. PMID: 3421013.

15. Saito H, Ono I, Ebihara S, Yoshizumi T, Ikeda S, Ono R. [Cases of head and neck carcinoma detected by sputum cytology-lung cancer screening and head and neck cancer]. Gan No Rinsho. 1985; 10. 31(13):1665–1668. Japanese. PMID: 4079057.

16. Jang SJ, Soria JC, Wang L, Hassan KA, Morice RC, Walsh GL, et al. Activation of melanoma antigen tumor antigens occurs early in lung carcinogenesis. Cancer Res. 2001; 11. 61(21):7959–7963. PMID: 11691819.

17. Bialkowska-Hobrzanska H, Bowles L, Bukala B, Joseph MG, Fletcher R, Razvi H. Comparison of human telomerase reverse transcriptase messenger RNA and telomerase activity as urine markers for diagnosis of bladder carcinoma. Mol Diagn. 2000; 12. 5(4):267–277. PMID: 11172490.

18. Piva MG, Navaglia F, Basso D, Fogar P, Roveroni G, Gallo N, et al. CEA mRNA identification in peripheral blood is feasible for colorectal, but not for gastric or pancreatic cancer staging. Oncology. 2000; 11. 59(4):323–328. PMID: 11096345.