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Identification of ATP synthase beta subunit (ATPB) on the cell surface as a non-small cell lung cancer (NSCLC) associated antigen.

Lu ZJ, Song QF, Jiang SS, Song Q, Wang W, Zhang GH, Kan B, Chen LJ, Yang JL, Luo F, Qian ZY, Wei YQ, Gou LT - BMC Cancer (2009)

Bottom Line: But the main problem is that only a few tumor-associated antigens or therapeutic targets have been known to us so far.The monoclonal antibody 4E7 (McAb4E7) specific against A549 cells was produced, which exhibited inhibitory effect on the proliferation of A549 cells.ATPB may be a potential biomarker and therapeutic target for the immunotherapy of NSCLC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, 37 Guoxue Xiang Street, Chengdu 610041, Sichuan, PR China. luzejun.01@163.com

ABSTRACT

Background: Antibody-based immunotherapy has achieved some success for cancer. But the main problem is that only a few tumor-associated antigens or therapeutic targets have been known to us so far. It is essential to identify more immunogenic antigens (especially cellular membrane markers) for tumor diagnosis and therapy.

Methods: The membrane proteins of lung adenocarcinoma cell line A549 were used to immunize the BALB/c mice. A monoclonal antibody 4E7 (McAb4E7) was produced with hybridoma technique. MTT cell proliferation assay was carried out to evaluate the inhibitory effect of McAb4E7 on A549 cells. Flow cytometric assay, immunohistochemistry, western blot and proteomic technologies based on 2-DE and mass spectrometry were employed to detect and identify the corresponding antigen of McAb4E7.

Results: The monoclonal antibody 4E7 (McAb4E7) specific against A549 cells was produced, which exhibited inhibitory effect on the proliferation of A549 cells. By the proteomic technologies, we identified that ATP synthase beta subunit (ATPB) was the corresponding antigen of McAb4E7. Then, flow cytometric analysis demonstrated the localization of the targeting antigen of McAb4E7 was on the A549 cells surface. Furthermore, immunohistochemistry showed that the antigen of McAb4E7 mainly aberrantly expressed in tumor cellular membrane in non-small cell lung cancer (NSCLC), but not in small cell lung cancer (SCLC). The rate of ectopic expressed ATPB in the cellular membrane in lung adenocarcinoma, squamous carcinoma and their adjacent nontumourous lung tissues was 71.88%, 66.67% and 25.81% respectively.

Conclusion: In the present study, we identified that the ectopic ATPB in tumor cellular membrane was the non-small cell lung cancer (NSCLC) associated antigen. ATPB may be a potential biomarker and therapeutic target for the immunotherapy of NSCLC.

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Subcellular localization of the antigen of McAb4E7. (A and B) The targeting antigen mainly located in the cellar membrane both in lung adenocarcinoma and squamous carcinoma (C) A little expression of the targeting antigen could be seen in the cytoplasm in the adjacent nontumourous lung tissues (D) The targeting antigen was not be found in the SCLC tissues; Magnifications: A-D × 40.
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Figure 3: Subcellular localization of the antigen of McAb4E7. (A and B) The targeting antigen mainly located in the cellar membrane both in lung adenocarcinoma and squamous carcinoma (C) A little expression of the targeting antigen could be seen in the cytoplasm in the adjacent nontumourous lung tissues (D) The targeting antigen was not be found in the SCLC tissues; Magnifications: A-D × 40.

Mentions: To further investigate the expression of the antigen of McAb4E7 in lung cancer tissues, we performed immunohistochemical analysis using paraffin-embedded tissue specimens. The expression of the antigen of McAb4E7 was obviously positive in the cellular membrane of lung adenocarcinoma and squamous carcinoma tissues (Figure 3A and 3B), while weak expression of the antigen of McAb4E7 could be seen in the cytoplasm in the adjacent nontumourous tissues of NSCLC (Figure 3C). The antigen of McAb4E7 was not be found in the tissues of SCLC (Figure 3D). The rate of expressed antigen of McAb4E7 in the cellular membrane in lung adenocarcinoma, squamous carcinoma and adjacent nontumourous lung tissues was 71.88%, 66.67% and 25.81% respectively (Table 1). The rate of membrane expressed antigen had significant difference between NSCLC tissues and their adjacent nontumourous tissues (P < 0.05), and it had no statistical difference between lung adenocarcinoma and squamous carcinoma (P > 0.05).


Identification of ATP synthase beta subunit (ATPB) on the cell surface as a non-small cell lung cancer (NSCLC) associated antigen.

Lu ZJ, Song QF, Jiang SS, Song Q, Wang W, Zhang GH, Kan B, Chen LJ, Yang JL, Luo F, Qian ZY, Wei YQ, Gou LT - BMC Cancer (2009)

Subcellular localization of the antigen of McAb4E7. (A and B) The targeting antigen mainly located in the cellar membrane both in lung adenocarcinoma and squamous carcinoma (C) A little expression of the targeting antigen could be seen in the cytoplasm in the adjacent nontumourous lung tissues (D) The targeting antigen was not be found in the SCLC tissues; Magnifications: A-D × 40.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2654462&req=5

Figure 3: Subcellular localization of the antigen of McAb4E7. (A and B) The targeting antigen mainly located in the cellar membrane both in lung adenocarcinoma and squamous carcinoma (C) A little expression of the targeting antigen could be seen in the cytoplasm in the adjacent nontumourous lung tissues (D) The targeting antigen was not be found in the SCLC tissues; Magnifications: A-D × 40.
Mentions: To further investigate the expression of the antigen of McAb4E7 in lung cancer tissues, we performed immunohistochemical analysis using paraffin-embedded tissue specimens. The expression of the antigen of McAb4E7 was obviously positive in the cellular membrane of lung adenocarcinoma and squamous carcinoma tissues (Figure 3A and 3B), while weak expression of the antigen of McAb4E7 could be seen in the cytoplasm in the adjacent nontumourous tissues of NSCLC (Figure 3C). The antigen of McAb4E7 was not be found in the tissues of SCLC (Figure 3D). The rate of expressed antigen of McAb4E7 in the cellular membrane in lung adenocarcinoma, squamous carcinoma and adjacent nontumourous lung tissues was 71.88%, 66.67% and 25.81% respectively (Table 1). The rate of membrane expressed antigen had significant difference between NSCLC tissues and their adjacent nontumourous tissues (P < 0.05), and it had no statistical difference between lung adenocarcinoma and squamous carcinoma (P > 0.05).

Bottom Line: But the main problem is that only a few tumor-associated antigens or therapeutic targets have been known to us so far.The monoclonal antibody 4E7 (McAb4E7) specific against A549 cells was produced, which exhibited inhibitory effect on the proliferation of A549 cells.ATPB may be a potential biomarker and therapeutic target for the immunotherapy of NSCLC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, 37 Guoxue Xiang Street, Chengdu 610041, Sichuan, PR China. luzejun.01@163.com

ABSTRACT

Background: Antibody-based immunotherapy has achieved some success for cancer. But the main problem is that only a few tumor-associated antigens or therapeutic targets have been known to us so far. It is essential to identify more immunogenic antigens (especially cellular membrane markers) for tumor diagnosis and therapy.

Methods: The membrane proteins of lung adenocarcinoma cell line A549 were used to immunize the BALB/c mice. A monoclonal antibody 4E7 (McAb4E7) was produced with hybridoma technique. MTT cell proliferation assay was carried out to evaluate the inhibitory effect of McAb4E7 on A549 cells. Flow cytometric assay, immunohistochemistry, western blot and proteomic technologies based on 2-DE and mass spectrometry were employed to detect and identify the corresponding antigen of McAb4E7.

Results: The monoclonal antibody 4E7 (McAb4E7) specific against A549 cells was produced, which exhibited inhibitory effect on the proliferation of A549 cells. By the proteomic technologies, we identified that ATP synthase beta subunit (ATPB) was the corresponding antigen of McAb4E7. Then, flow cytometric analysis demonstrated the localization of the targeting antigen of McAb4E7 was on the A549 cells surface. Furthermore, immunohistochemistry showed that the antigen of McAb4E7 mainly aberrantly expressed in tumor cellular membrane in non-small cell lung cancer (NSCLC), but not in small cell lung cancer (SCLC). The rate of ectopic expressed ATPB in the cellular membrane in lung adenocarcinoma, squamous carcinoma and their adjacent nontumourous lung tissues was 71.88%, 66.67% and 25.81% respectively.

Conclusion: In the present study, we identified that the ectopic ATPB in tumor cellular membrane was the non-small cell lung cancer (NSCLC) associated antigen. ATPB may be a potential biomarker and therapeutic target for the immunotherapy of NSCLC.

Show MeSH
Related in: MedlinePlus