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Interleukin-6 (IL-6) as an autocrine growth factor in oral squamous cell carcinoma In vitro
146
P27 - Poster session: Tuesday, April 27, 1999
24. OK-432 Conjugated Tumor Vaccine Induces Tumor-Specific Immunity for SCC of the Tongue
25. Interleukin-6 (IL-6) as an Autocrine Growth Factor in Oral Squamous Cell Carcinoma In Vitro
Bukawa, 1t., Tsuyuki, Y, Li, X., Kawabe, R., Omura, S., Chikumaru, H., Mizuki, N., Aoki, S., Fufita, K.
lmai, ya, Sasaki, T..a, Akimoto, K. 2, Niwa, A. 3, Fujibayashi, T.1
Department or Oral Maxillofacial Surgery, Yokohama City University School of Medicine, Yokohama, Japan
1Department of Oral and Maxillofacial Surgery, 2Laboratory of Molecular and Cellular Biology, 3Department of Microbiology, Dokkyo University School of Medicine, Tochigi, Japan
Purpose To induce tumor-specific immunity for tumor inoculated into tongue of mouse by immunization with a newly developed tumor vaccine. Methods Tumor cells (SCC from mouse) were mixed with OK-432 which was a lyophilized streptococcal preparation (Picibanil; Chugal Pharm. Co., Ltd., Japan), and conjugated each other using a glutaraldehyde (GA) method as previously described (Bur~AWA et al. Nature Med 1995: 1: 681). 1, 8 and 15 days after tumor cells were inoculated into tongue of mice, mice were immunized with OK-432 conjugated tumor vaccine or saline. Tumor incidence and mortality of immunized mice were compared to control. Results OK-432 conjugated tumor vaccine was observed like star form 30 minutes after conjugation. This connection was so stable that did not separate each other by 3 times of centrifuges in order to remove surplus G A and untreated OK-432 from conjugated tumor cells. We made sure of the safety of this OK-432 conjugated vaccine as follows: About 80% of the conjugated cells were stained with trypan blue, indicating that many OK-432 conjugated tumor cells were not alive. Culture of the OK-432 conjugated tumor cells induced no proliferation of tumor cells. When mice were immunized 3 times with OK-432 conjugated tumor vaccine,-suppression of tumor incidence was observed in comparison with control mice. Mortality of mice was also improved by immunization with OK-432 conjugated tumor vaccine. Conclusions OK-432 conjugated tumor vaccine, which was newly constructed, may have a feasibility to induce tumor-specific immunity for SCC inoculated into tongue of mouse.
Purpose Recent studies have reported that Interleukin-6 (IL-6) is secreted by tumor cells and stimulates the]r growth in vitro. However, there are no reports available of the performance of IL-6 in oral cancer. We conducted the present study to define the role of IL-6 in oral squamous cell carcinoma. Material and methods A squamous cell carcinoma cell line (STC) derived from a tongue cancer was used. The content o f IL-6 in the conditioned medium (CM) was measured by means of an enzyme-linked immunosorbent assay (ELISA). Expression of IL-6/IL-6R m R N A was detected by the reverse transcription PCR (RT-PCR) method. The effects of TNF-c~ (1~100 ng/ ml), IL-1 (1-100 ng/ml) or LPS (10-1000 ng/ml) on IL-6 secretion and expression of IL-6 m R N A by the cells were examined by E L I S A and northern blot analysis. Furthermore, the effects of IL-6 (100 pg/ml) or anti-IL-6 antibody (1-100 gg/ml) on cell growth of the cells were examined. Results The STC cells secreted IL-6 into the CM (2440 pg/ml/72 h) and expressed IL-6 (295 bp) and IL-6R m R N A (gp 80:251 bp/gp 130:423 bp). The STC cells treated with T N F - a (1000 ng/ml) increased IL-6 m R N A ' i n the cells and the concentration of IL-6 in-the CM (77200 pg/ml/24 h). Cell growth of the cells was stimulated by IL-6 (P=0.006), while anti-IL6 antibody inhibited growth (P=0.0341). Conclusion These findings suggest that the IL-6 produced by the STC cells seems to act as an autocrine growth factor.
26. Human DOC-1 Suppression of Cell Growth and Association with the Monomeric Form of CDK2
Shintani, S., Nakahara, Y, Tsuji, T., McBride, J., Todd, R., Ohyama, H., Wong, Do Division of Oral Pathology, Department of Oral Medicine and Diagnostic Science, Harvard School of Dental Medicine DOC-1, deleted in oral cancer-l, originally isolated from hamster oral keratinocytes due to its differential expression in normal keratinocyte. Transfection of hamster DOC-1 c D N A can suppress some transformed phenotypes of malignant oral keratinocyte in hamster model. Human DOC-1
P27 - Poster session: Tuesday, April 27, 1999
24. OK-432 Conjugated Tumor Vaccine Induces Tumor-Specific Immunity for SCC of the Tongue
25. Interleukin-6 (IL-6) as an Autocrine Growth Factor in Oral Squamous Cell Carcinoma In Vitro
Bukawa, 1t., Tsuyuki, Y, Li, X., Kawabe, R., Omura, S., Chikumaru, H., Mizuki, N., Aoki, S., Fufita, K.
lmai, ya, Sasaki, T..a, Akimoto, K. 2, Niwa, A. 3, Fujibayashi, T.1
Department or Oral Maxillofacial Surgery, Yokohama City University School of Medicine, Yokohama, Japan
1Department of Oral and Maxillofacial Surgery, 2Laboratory of Molecular and Cellular Biology, 3Department of Microbiology, Dokkyo University School of Medicine, Tochigi, Japan
Purpose To induce tumor-specific immunity for tumor inoculated into tongue of mouse by immunization with a newly developed tumor vaccine. Methods Tumor cells (SCC from mouse) were mixed with OK-432 which was a lyophilized streptococcal preparation (Picibanil; Chugal Pharm. Co., Ltd., Japan), and conjugated each other using a glutaraldehyde (GA) method as previously described (Bur~AWA et al. Nature Med 1995: 1: 681). 1, 8 and 15 days after tumor cells were inoculated into tongue of mice, mice were immunized with OK-432 conjugated tumor vaccine or saline. Tumor incidence and mortality of immunized mice were compared to control. Results OK-432 conjugated tumor vaccine was observed like star form 30 minutes after conjugation. This connection was so stable that did not separate each other by 3 times of centrifuges in order to remove surplus G A and untreated OK-432 from conjugated tumor cells. We made sure of the safety of this OK-432 conjugated vaccine as follows: About 80% of the conjugated cells were stained with trypan blue, indicating that many OK-432 conjugated tumor cells were not alive. Culture of the OK-432 conjugated tumor cells induced no proliferation of tumor cells. When mice were immunized 3 times with OK-432 conjugated tumor vaccine,-suppression of tumor incidence was observed in comparison with control mice. Mortality of mice was also improved by immunization with OK-432 conjugated tumor vaccine. Conclusions OK-432 conjugated tumor vaccine, which was newly constructed, may have a feasibility to induce tumor-specific immunity for SCC inoculated into tongue of mouse.
Purpose Recent studies have reported that Interleukin-6 (IL-6) is secreted by tumor cells and stimulates the]r growth in vitro. However, there are no reports available of the performance of IL-6 in oral cancer. We conducted the present study to define the role of IL-6 in oral squamous cell carcinoma. Material and methods A squamous cell carcinoma cell line (STC) derived from a tongue cancer was used. The content o f IL-6 in the conditioned medium (CM) was measured by means of an enzyme-linked immunosorbent assay (ELISA). Expression of IL-6/IL-6R m R N A was detected by the reverse transcription PCR (RT-PCR) method. The effects of TNF-c~ (1~100 ng/ ml), IL-1 (1-100 ng/ml) or LPS (10-1000 ng/ml) on IL-6 secretion and expression of IL-6 m R N A by the cells were examined by E L I S A and northern blot analysis. Furthermore, the effects of IL-6 (100 pg/ml) or anti-IL-6 antibody (1-100 gg/ml) on cell growth of the cells were examined. Results The STC cells secreted IL-6 into the CM (2440 pg/ml/72 h) and expressed IL-6 (295 bp) and IL-6R m R N A (gp 80:251 bp/gp 130:423 bp). The STC cells treated with T N F - a (1000 ng/ml) increased IL-6 m R N A ' i n the cells and the concentration of IL-6 in-the CM (77200 pg/ml/24 h). Cell growth of the cells was stimulated by IL-6 (P=0.006), while anti-IL6 antibody inhibited growth (P=0.0341). Conclusion These findings suggest that the IL-6 produced by the STC cells seems to act as an autocrine growth factor.
26. Human DOC-1 Suppression of Cell Growth and Association with the Monomeric Form of CDK2
Shintani, S., Nakahara, Y, Tsuji, T., McBride, J., Todd, R., Ohyama, H., Wong, Do Division of Oral Pathology, Department of Oral Medicine and Diagnostic Science, Harvard School of Dental Medicine DOC-1, deleted in oral cancer-l, originally isolated from hamster oral keratinocytes due to its differential expression in normal keratinocyte. Transfection of hamster DOC-1 c D N A can suppress some transformed phenotypes of malignant oral keratinocyte in hamster model. Human DOC-1