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The study on newly developed McAb NJ001 specific to non-small cell lung cancer and its biological characteristics.

Pan S, Wang F, Huang P, Xu T, Zhang L, Xu J, Li Q, Xia W, Sun R, Huang L, Peng Y, Qin X, Shu Y, Hu Z, Shen H - PLoS ONE (2012)

Bottom Line: In soft agar assay, the colony formation efficiency in NJ001 groups decreased in a dose-dependent manner.Meanwhile, NJ001 caused significant reduction in tumor volume and tumor weight compared to control mice in lung cancer xenograft model.NJ001 also led to cytomorphological changes and induced the apoptosis of human lung adenocarcinoma cell line SPC-A1 significantly.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China. sypan@njmu.edu.cn

ABSTRACT
Monoclonal antibody (McAb) is the key tool for cancer immunodiagnosis and immunotherapy. McAb-based immunotherapy that targets tumor antigens has had great achivement. In this study, a cell clone which kept secreting high-titer IgG1-type McAb named NJ001 against human non-small cell lung cancer (NSCLC) cells was obtained. The titer of purified NJ001 was 2×10(6). The antigen named SP70 of NSCLC specifically identified by NJ001 was proved to be a protein with the relative molecular mass (Mr) of 70 kDa. The results of immunohistochemical staining indicated that NJ001 could positively react to NSCLC, but weak positively or negatively react to human small-cell lung cancer (SCLC), pulmonary pseudotumor and other epithelial tumors. In soft agar assay, the colony formation efficiency in NJ001 groups decreased in a dose-dependent manner. For the concentration of 100 µg/ml, 200 µg/ml and 400 µg/ml, the inhibition ratio of colony formation was 23.4%, 62.5% and 100% respectively. Meanwhile, NJ001 caused significant reduction in tumor volume and tumor weight compared to control mice in lung cancer xenograft model. The tumor growth inhibition ratio in 200 µg, 400 µg and 800 µg NJ001 groups was 10.44%, 37.29% and 44.04%, respectively. NJ001 also led to cytomorphological changes and induced the apoptosis of human lung adenocarcinoma cell line SPC-A1 significantly. The newly developed NJ001 selectively reacted to NSCLC and exhibited anti-tumor activity both in vitro and in vivo. NJ001 is of great value concerning immunodiagnostics and immunotherapy for NSCLC and holds promise for further research regarding the mechanism underlying tumor progression of NSCLC.

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Inhibition of colony formation of SPC-A1 cells by NJ001 in soft agar. (×100).The cell suspensions (2×104 cells) mixed with 0.3% agarose and different concentrations of NJ001 or MCA2849 were layered on the top of culture media in 6-well culture plates and allowed to grow for 2 weeks before colonies were counted. Representative contrast images were shown. (A) 0 µg/mL NJ001, (B) 100 µg/mL NJ001, (C) 200 µg/mL NJ001, (D) 400 µg/mL NJ001, (E) 0 µg/mL MCA2849, (F) 100 µg/mL MCA2849, (G) 200 µg/mL MCA2849, (H) 400 µg/mL MCA2849.
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pone-0033009-g004: Inhibition of colony formation of SPC-A1 cells by NJ001 in soft agar. (×100).The cell suspensions (2×104 cells) mixed with 0.3% agarose and different concentrations of NJ001 or MCA2849 were layered on the top of culture media in 6-well culture plates and allowed to grow for 2 weeks before colonies were counted. Representative contrast images were shown. (A) 0 µg/mL NJ001, (B) 100 µg/mL NJ001, (C) 200 µg/mL NJ001, (D) 400 µg/mL NJ001, (E) 0 µg/mL MCA2849, (F) 100 µg/mL MCA2849, (G) 200 µg/mL MCA2849, (H) 400 µg/mL MCA2849.

Mentions: SPC-A1 cells were plated on a soft agar matrix, treated with NJ001 or MCA2849 (irrelevant McAb) (0, 100, 200, 400, 800, or 1000 µg/mL) and incubated at the condition of 37°C with 5% CO2. After 14 days, the number of colonies was counted and the representative images were obtained (Figure 4). As shown in Table 2, NJ001 inhibited colony formation in a dose-dependent manner, exhibiting 23.4% inhibition ratio at 100 µg/mL, 62.5% inhibition ratio at 200 µg/mL. When the concentration reached 400 µg/mL or higher, there were no colonies larger than 50 cells, showing a 100% inhibition ratio. However, MCA2849 didn't inhibit the colony formation of SPC-A1.


The study on newly developed McAb NJ001 specific to non-small cell lung cancer and its biological characteristics.

Pan S, Wang F, Huang P, Xu T, Zhang L, Xu J, Li Q, Xia W, Sun R, Huang L, Peng Y, Qin X, Shu Y, Hu Z, Shen H - PLoS ONE (2012)

Inhibition of colony formation of SPC-A1 cells by NJ001 in soft agar. (×100).The cell suspensions (2×104 cells) mixed with 0.3% agarose and different concentrations of NJ001 or MCA2849 were layered on the top of culture media in 6-well culture plates and allowed to grow for 2 weeks before colonies were counted. Representative contrast images were shown. (A) 0 µg/mL NJ001, (B) 100 µg/mL NJ001, (C) 200 µg/mL NJ001, (D) 400 µg/mL NJ001, (E) 0 µg/mL MCA2849, (F) 100 µg/mL MCA2849, (G) 200 µg/mL MCA2849, (H) 400 µg/mL MCA2849.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3316548&req=5

pone-0033009-g004: Inhibition of colony formation of SPC-A1 cells by NJ001 in soft agar. (×100).The cell suspensions (2×104 cells) mixed with 0.3% agarose and different concentrations of NJ001 or MCA2849 were layered on the top of culture media in 6-well culture plates and allowed to grow for 2 weeks before colonies were counted. Representative contrast images were shown. (A) 0 µg/mL NJ001, (B) 100 µg/mL NJ001, (C) 200 µg/mL NJ001, (D) 400 µg/mL NJ001, (E) 0 µg/mL MCA2849, (F) 100 µg/mL MCA2849, (G) 200 µg/mL MCA2849, (H) 400 µg/mL MCA2849.
Mentions: SPC-A1 cells were plated on a soft agar matrix, treated with NJ001 or MCA2849 (irrelevant McAb) (0, 100, 200, 400, 800, or 1000 µg/mL) and incubated at the condition of 37°C with 5% CO2. After 14 days, the number of colonies was counted and the representative images were obtained (Figure 4). As shown in Table 2, NJ001 inhibited colony formation in a dose-dependent manner, exhibiting 23.4% inhibition ratio at 100 µg/mL, 62.5% inhibition ratio at 200 µg/mL. When the concentration reached 400 µg/mL or higher, there were no colonies larger than 50 cells, showing a 100% inhibition ratio. However, MCA2849 didn't inhibit the colony formation of SPC-A1.

Bottom Line: In soft agar assay, the colony formation efficiency in NJ001 groups decreased in a dose-dependent manner.Meanwhile, NJ001 caused significant reduction in tumor volume and tumor weight compared to control mice in lung cancer xenograft model.NJ001 also led to cytomorphological changes and induced the apoptosis of human lung adenocarcinoma cell line SPC-A1 significantly.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China. sypan@njmu.edu.cn

ABSTRACT
Monoclonal antibody (McAb) is the key tool for cancer immunodiagnosis and immunotherapy. McAb-based immunotherapy that targets tumor antigens has had great achivement. In this study, a cell clone which kept secreting high-titer IgG1-type McAb named NJ001 against human non-small cell lung cancer (NSCLC) cells was obtained. The titer of purified NJ001 was 2×10(6). The antigen named SP70 of NSCLC specifically identified by NJ001 was proved to be a protein with the relative molecular mass (Mr) of 70 kDa. The results of immunohistochemical staining indicated that NJ001 could positively react to NSCLC, but weak positively or negatively react to human small-cell lung cancer (SCLC), pulmonary pseudotumor and other epithelial tumors. In soft agar assay, the colony formation efficiency in NJ001 groups decreased in a dose-dependent manner. For the concentration of 100 µg/ml, 200 µg/ml and 400 µg/ml, the inhibition ratio of colony formation was 23.4%, 62.5% and 100% respectively. Meanwhile, NJ001 caused significant reduction in tumor volume and tumor weight compared to control mice in lung cancer xenograft model. The tumor growth inhibition ratio in 200 µg, 400 µg and 800 µg NJ001 groups was 10.44%, 37.29% and 44.04%, respectively. NJ001 also led to cytomorphological changes and induced the apoptosis of human lung adenocarcinoma cell line SPC-A1 significantly. The newly developed NJ001 selectively reacted to NSCLC and exhibited anti-tumor activity both in vitro and in vivo. NJ001 is of great value concerning immunodiagnostics and immunotherapy for NSCLC and holds promise for further research regarding the mechanism underlying tumor progression of NSCLC.

Show MeSH
Related in: MedlinePlus