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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

NAP-1 protein is a secreted protein secreted by FDC. A: NAP-1 gene is expressed in S100+ CD35+ FDC cell lines 1. In situ hybridization by Dig labelled NAP-1 cDNA probe in FDC lines (×300); 2. Immunohistochemistry by Mouse Anti-S100 Monoclonal antibody in FDC lines (×300); 3. Immunohistochemistry by Mouse Anti-CD35 Monoclonal antibody in FDC lines (×300); 4. Immunohistochemistry by Rabbit Anti-NAP-1 polyclonal antibody in FDC lines (×300) B: Western blotting analysis of FDC culture supernatants using rabbit anti-NAP-1 polyclonal antibody 1. The negative antibody control: the primary antibody replaced with block solution; 2. The negative supernatants control: the FDC lines culture supernatants replaced with fresh media; 3. Western blotting analysis of FDC supernatants cultured for 1 day; 4. Western blotting analysis of FDC supernatants cultured for 2 days
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Figure 3: NAP-1 protein is a secreted protein secreted by FDC. A: NAP-1 gene is expressed in S100+ CD35+ FDC cell lines 1. In situ hybridization by Dig labelled NAP-1 cDNA probe in FDC lines (×300); 2. Immunohistochemistry by Mouse Anti-S100 Monoclonal antibody in FDC lines (×300); 3. Immunohistochemistry by Mouse Anti-CD35 Monoclonal antibody in FDC lines (×300); 4. Immunohistochemistry by Rabbit Anti-NAP-1 polyclonal antibody in FDC lines (×300) B: Western blotting analysis of FDC culture supernatants using rabbit anti-NAP-1 polyclonal antibody 1. The negative antibody control: the primary antibody replaced with block solution; 2. The negative supernatants control: the FDC lines culture supernatants replaced with fresh media; 3. Western blotting analysis of FDC supernatants cultured for 1 day; 4. Western blotting analysis of FDC supernatants cultured for 2 days

Mentions: In situ hybridization and immunohistochemistry suggested that the NAP-1 peptide is expressed in S100+ CD35+ FDC of the germinal center and not in other stromal immune cells in NPC and NP tissues (Figure 2). RT-PCR, Western Blotting, immunohistochemistry and in situ hybridization of FDC lines further demonstrated that NAP-1 is a protein specifically secreted by these cells (Figure 3).


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)

NAP-1 protein is a secreted protein secreted by FDC. A: NAP-1 gene is expressed in S100+ CD35+ FDC cell lines 1. In situ hybridization by Dig labelled NAP-1 cDNA probe in FDC lines (×300); 2. Immunohistochemistry by Mouse Anti-S100 Monoclonal antibody in FDC lines (×300); 3. Immunohistochemistry by Mouse Anti-CD35 Monoclonal antibody in FDC lines (×300); 4. Immunohistochemistry by Rabbit Anti-NAP-1 polyclonal antibody in FDC lines (×300) B: Western blotting analysis of FDC culture supernatants using rabbit anti-NAP-1 polyclonal antibody 1. The negative antibody control: the primary antibody replaced with block solution; 2. The negative supernatants control: the FDC lines culture supernatants replaced with fresh media; 3. Western blotting analysis of FDC supernatants cultured for 1 day; 4. Western blotting analysis of FDC supernatants cultured for 2 days
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Related In: Results  -  Collection

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

Figure 3: NAP-1 protein is a secreted protein secreted by FDC. A: NAP-1 gene is expressed in S100+ CD35+ FDC cell lines 1. In situ hybridization by Dig labelled NAP-1 cDNA probe in FDC lines (×300); 2. Immunohistochemistry by Mouse Anti-S100 Monoclonal antibody in FDC lines (×300); 3. Immunohistochemistry by Mouse Anti-CD35 Monoclonal antibody in FDC lines (×300); 4. Immunohistochemistry by Rabbit Anti-NAP-1 polyclonal antibody in FDC lines (×300) B: Western blotting analysis of FDC culture supernatants using rabbit anti-NAP-1 polyclonal antibody 1. The negative antibody control: the primary antibody replaced with block solution; 2. The negative supernatants control: the FDC lines culture supernatants replaced with fresh media; 3. Western blotting analysis of FDC supernatants cultured for 1 day; 4. Western blotting analysis of FDC supernatants cultured for 2 days
Mentions: In situ hybridization and immunohistochemistry suggested that the NAP-1 peptide is expressed in S100+ CD35+ FDC of the germinal center and not in other stromal immune cells in NPC and NP tissues (Figure 2). RT-PCR, Western Blotting, immunohistochemistry and in situ hybridization of FDC lines further demonstrated that NAP-1 is a protein specifically secreted by these cells (Figure 3).

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