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Identification and characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1.

Li F, Yang XY, Jiang WH, Yin ZH, Feng XL, Liu WD, Wang L, Zhou W, Ren CP, Yao KT - J Transl Med (2004)

Bottom Line: Several NPC-associated genes have been so far described and here we describe the identification and the characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1.Surprisingly, differential RT-PCR demonstrated decreased expression of NAP-1 in NPC compared with paired NP biopsies in 42.5 % of cases (17 out of 40).Therefore, it is likely that NAP-1 is secreted by FDC in the NP and may play an immune modulatory role in NPC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, China, 410078. ktyao@fimmu.com

ABSTRACT
Nasopharyngeal carcinoma (NPC) is one of the most commons cancers in Southeast Asia and Southern China. Several NPC-associated genes have been so far described and here we describe the identification and the characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1. NAP-1 was identified with the human genome draft searching method combined with nested PCR mapping of the chromosome 4q13 region. NAP-1 encodes an 85 amino acid alkaline peptide with a calculated isoelectric point of 9.3, three phosphorilation sites and a proline-rich region. Northern blot analysis revealed that NAP-1 is expressed as a 0.6 kb transcript in normal lymph nodes and trachea. In addition, reverse transcription (RT)-PCR showed that NAP-1 is expressed not only in NPC but in normal nasopharynx (NP) and various other tumors and tissues of the head and neck including: tonsils, lymph nodes, carcinoma of the tonsil, T cell lymphomas, squamous cell carcinoma of the hard palate, papilloma of the nasopharynx, nasopharyngitis, lymphoma of the tongue root and follicular dendritic cells (FDC). In addition, NAP-1 is not expressed in normal tissues or tumors from other anatomical regions and was not expressed by NPC cell lines. Surprisingly, differential RT-PCR demonstrated decreased expression of NAP-1 in NPC compared with paired NP biopsies in 42.5 % of cases (17 out of 40). In addition, in situ hybridization and immunohistochemistry demonstrated that NAP-1 is expressed by S100+ CD35+ FDCs of the germinal center and not in other normal immune cells infiltrating NP or NPC. Therefore, it is likely that NAP-1 is secreted by FDC in the NP and may play an immune modulatory role in NPC.

No MeSH data available.


Related in: MedlinePlus

Cloning and identification of NAP-1 gene A: Identification of 5' end's PCR product of the gene by using human genome draft searching methodcombined with nested PCR 1. pFRW-2 and pRVS-3(no); 2. pFRW-2 and pRVS-2(no); 3. pFRW-3 and pRVS-3(176 bp); 4. pFRW-3 and pRVS-2(282 bp) M: DNA ladder B: Northern blots analysis of the gene in 15 kinds of human tissues 1. heart; 2. brain; 3. placenta; 4. lung; 5. liver; 6. skeletal muscle; 7. kidney; 8. pancreas; 9. stomach; 10. thyroid; 11. spinal cord; 12. lymph node; 13. trachea; 14. adrenal gland; 15. bone marrow
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Figure 1: Cloning and identification of NAP-1 gene A: Identification of 5' end's PCR product of the gene by using human genome draft searching methodcombined with nested PCR 1. pFRW-2 and pRVS-3(no); 2. pFRW-2 and pRVS-2(no); 3. pFRW-3 and pRVS-3(176 bp); 4. pFRW-3 and pRVS-2(282 bp) M: DNA ladder B: Northern blots analysis of the gene in 15 kinds of human tissues 1. heart; 2. brain; 3. placenta; 4. lung; 5. liver; 6. skeletal muscle; 7. kidney; 8. pancreas; 9. stomach; 10. thyroid; 11. spinal cord; 12. lymph node; 13. trachea; 14. adrenal gland; 15. bone marrow

Mentions: Therefore, we focused our attention an the latter gene and an EST contig (502 bp) was assembled by combining information from 4 highly homologous ESTs (GenBank accession number CB140082, AI332560, BG059093 and AI701589) according to EST BLAST searches using a cDNA fragment (GenBank accession number BG231197) as electronic probe. Electronic prolongation by EST contig did not reveal a complete open reading frame. Bioinformatics analysis suggested the contig resembled a human peptide precursor secreted by follicular dendritic cells (FDC-SP) (GenBank accession number AF435080) [10] with a 77% homology. No stop codons in frame with the cDNA 5' end start codon of the FDC-SP could be identified. The results suggested that the full-length cDNAs could be obtained by combining the Human Genome Draft Searching Method with nested PCR (Figure 1a). Northern blot analysis revealed that the gene produced a transcript (0.6 kb) that could be identified among normal tissues only in lymph nodes and trachea (Figure 1b).


Identification and characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1.

Li F, Yang XY, Jiang WH, Yin ZH, Feng XL, Liu WD, Wang L, Zhou W, Ren CP, Yao KT - J Transl Med (2004)

Cloning and identification of NAP-1 gene A: Identification of 5' end's PCR product of the gene by using human genome draft searching methodcombined with nested PCR 1. pFRW-2 and pRVS-3(no); 2. pFRW-2 and pRVS-2(no); 3. pFRW-3 and pRVS-3(176 bp); 4. pFRW-3 and pRVS-2(282 bp) M: DNA ladder B: Northern blots analysis of the gene in 15 kinds of human tissues 1. heart; 2. brain; 3. placenta; 4. lung; 5. liver; 6. skeletal muscle; 7. kidney; 8. pancreas; 9. stomach; 10. thyroid; 11. spinal cord; 12. lymph node; 13. trachea; 14. adrenal gland; 15. bone marrow
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC411061&req=5

Figure 1: Cloning and identification of NAP-1 gene A: Identification of 5' end's PCR product of the gene by using human genome draft searching methodcombined with nested PCR 1. pFRW-2 and pRVS-3(no); 2. pFRW-2 and pRVS-2(no); 3. pFRW-3 and pRVS-3(176 bp); 4. pFRW-3 and pRVS-2(282 bp) M: DNA ladder B: Northern blots analysis of the gene in 15 kinds of human tissues 1. heart; 2. brain; 3. placenta; 4. lung; 5. liver; 6. skeletal muscle; 7. kidney; 8. pancreas; 9. stomach; 10. thyroid; 11. spinal cord; 12. lymph node; 13. trachea; 14. adrenal gland; 15. bone marrow
Mentions: Therefore, we focused our attention an the latter gene and an EST contig (502 bp) was assembled by combining information from 4 highly homologous ESTs (GenBank accession number CB140082, AI332560, BG059093 and AI701589) according to EST BLAST searches using a cDNA fragment (GenBank accession number BG231197) as electronic probe. Electronic prolongation by EST contig did not reveal a complete open reading frame. Bioinformatics analysis suggested the contig resembled a human peptide precursor secreted by follicular dendritic cells (FDC-SP) (GenBank accession number AF435080) [10] with a 77% homology. No stop codons in frame with the cDNA 5' end start codon of the FDC-SP could be identified. The results suggested that the full-length cDNAs could be obtained by combining the Human Genome Draft Searching Method with nested PCR (Figure 1a). Northern blot analysis revealed that the gene produced a transcript (0.6 kb) that could be identified among normal tissues only in lymph nodes and trachea (Figure 1b).

Bottom Line: Several NPC-associated genes have been so far described and here we describe the identification and the characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1.Surprisingly, differential RT-PCR demonstrated decreased expression of NAP-1 in NPC compared with paired NP biopsies in 42.5 % of cases (17 out of 40).Therefore, it is likely that NAP-1 is secreted by FDC in the NP and may play an immune modulatory role in NPC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, China, 410078. ktyao@fimmu.com

ABSTRACT
Nasopharyngeal carcinoma (NPC) is one of the most commons cancers in Southeast Asia and Southern China. Several NPC-associated genes have been so far described and here we describe the identification and the characterization of a novel nasopharyngeal carcinoma-associated peptide: NAP-1. NAP-1 was identified with the human genome draft searching method combined with nested PCR mapping of the chromosome 4q13 region. NAP-1 encodes an 85 amino acid alkaline peptide with a calculated isoelectric point of 9.3, three phosphorilation sites and a proline-rich region. Northern blot analysis revealed that NAP-1 is expressed as a 0.6 kb transcript in normal lymph nodes and trachea. In addition, reverse transcription (RT)-PCR showed that NAP-1 is expressed not only in NPC but in normal nasopharynx (NP) and various other tumors and tissues of the head and neck including: tonsils, lymph nodes, carcinoma of the tonsil, T cell lymphomas, squamous cell carcinoma of the hard palate, papilloma of the nasopharynx, nasopharyngitis, lymphoma of the tongue root and follicular dendritic cells (FDC). In addition, NAP-1 is not expressed in normal tissues or tumors from other anatomical regions and was not expressed by NPC cell lines. Surprisingly, differential RT-PCR demonstrated decreased expression of NAP-1 in NPC compared with paired NP biopsies in 42.5 % of cases (17 out of 40). In addition, in situ hybridization and immunohistochemistry demonstrated that NAP-1 is expressed by S100+ CD35+ FDCs of the germinal center and not in other normal immune cells infiltrating NP or NPC. Therefore, it is likely that NAP-1 is secreted by FDC in the NP and may play an immune modulatory role in NPC.

No MeSH data available.


Related in: MedlinePlus