| Co-Investigator(Kenkyū-buntansha) |
HINOHARA Tadashi DOKKYO UNIVERSITY SCHOOL OF MEDICINE, OTORHINOLARYNGOLOGY & BRONCHOESOPHAGOLOGY, PROFESSOR, 医学部, 教授 (70049146)
UNO Kouhei DOKKYO UNIVERSITY SCHOOL OF MEDICINE, OTORHINOLARYNGOLOGY & BRONCHOESOPHAGOLOGY, LECTURER, 医学部, 講師 (50185055)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Research Abstract |
We investigated the effects of the TNF-α and INF-γ on the growth of the human squamous cell carcinoma of the head and neck (SCCHN) cell lines. To elucidate the mechanism of growth suppression, the expression of the TNF-α receptor and Fas antigen were measured by flow cytometry, and the rate of apoptotic cell death was measured via the TdT-mediated dUPTP-biotin nick labeling method on SCCHN cell line. TNF-α receptor were detected on the cell surface after 2h of incubation with TNF-α and INF-γ, either alone or in combination. Fas antigens were detected after 4, 7 and 14h of incubation. DNA fragmentation was demonstrated after 24h. These results suggest that the antiproliferative activity on TNF-α and INF-γ an SCCHN cell line is mediated by apoptosis. The human IL-2 gene is transduced into SCCHN cell lines to determine their biological activities. SCCHN cell lines were transduced of the DFG-hIL-2-Neo vector which contains a human IL-2 gene and a neomycin resistance (Neo) gene, used as a s
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electable marker. The phenotypic changes of PBLs were observed which were cultured with supernatants of parental and IL-2 gene-transduced PCI-13 cell lines. For in vivo studies, parental SCCHN cell line and IL-2 gene-transduced SCCHN cell line were injected s.c. into nude mice and measured with calipers every other day. The IL-2 gene-transduced PCI-13 cells secreted high doses of IL-2 12 mouths following transduction. Macrophages and NK cells were also more highly stimulated by IL-2 gene-transduced PCI-13 supernatant than by parental PCI-13 supernatant alone. The parental SCCHN cells injected subcutaneously into the flank of nude mice grew progressively. In contrast, IL-2 gene-transduced SCCHN cells grew initially after subcutaneous injection but were rejected after 8-14 days. The subcutaneous tumors had consistently regressed by day 28. The effectors of the SCCHN on immune cells and immune activated cells may be therapeutically useful, as suggested by our in vivo model and in vitro in our experimental results. Squamous cell carcinoma of the Head and Neck(SCCHN) cell surface antigens were identified by several monoclonal antibodies. These cell surface antigens might relate not only recognition of immune cells but also cell to cell adhesion. The SCCHN cell lines expressed squamous epithelial cell antigen (A9, E7, K928, K931) : mean positive percent of A9, 96.8%, E7, 96.2%, K931, 71.6%, K928, 94.2%. Except PCI-101 and PCI-104 showed low level (11.3% and 35.7%) of K931 antigen expression. In contrast, HLA-DR (class II) antigen only defined by PCI-51(82.3%). Also ICAM-1 which is adhesion molecule defined by PCI-51 (70.7%), PCI-107 (45.9%) and PCI-13 (23.2%). HLA class I antigen defined by all cell lines : mean positive percent is 83.9%. These result sugested the SCCHN cell lines expressed several kinds of antigens on these cell surface. These antigen might be used as targets for immune recognition, detection, and tumor cell destruction. Less
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