• Radiation Oncology Journal
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• v.19 no.2
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• pp.171-180
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• 2001

Purpose : Expression of TIMP, intrinsic inhibitor of MMP, is regulated by signal transduction in response to genotoxins and is likely to be an important step in metastasis, angiogenesis and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP expression and its mechanism in head and neck cancer cell lines. Materials and Methods : Human head and neck cancer cell lines established at Asan Medical Center were used and radiosensitivity $(D_0)$, radiation cytotoxicity and metastatic potential were measured by clonogenic assay, n assay and invasion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and expression of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promoter region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC-HN-1 and -HN-9 cells with or without expression vector Ras, then the cells were exposed to radiation or PMA, PKC activator. EMSA was peformed with oligonucleotide (-59/-53 element and SP1) of TIMP1 promoter. Results : $D_0$ of HN-1, -2, -3, -5 and -9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. n assay confirmed cell viability, over $94\%$ at 24hrs, 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuously. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN-1 and HN-9 cell lines but after 10 Gy irradiation, it was increased in all cell lines. At 48hrs after irradiation, it was increased in HN-1 but decreased in HN-9 cells. But the change in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN-1 was induced by PMA but in HN-9 cell lines, it was suppressed. Wild type Ras induced the TIMP-1 transcription by 20 fold and 4 fold in HN-1 and HN-9 respectively. The binding activity to -59/-53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines. Conclusions : We observed the difference of expression and activity of TIMPs between radiosensitive and radioresistant cell line and the different signal transduction pathway between in these cell lines may contribute the different radiosensitivity. Further research to investigate the radiation response and its signal pathway of TIMPs is needed.

• Korean Journal of Agricultural Science
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• v.2 no.1
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• pp.9-47
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• 1975

These experiments were caried out to study the effects of caponization and various hormone treatments upon meat production and improvement of meat quality of growing chicken. Sixtyseven days old 160 New Hampshire cockerels were treated and growth rate, carcass yield, change of weight of individual organs, meat composition and change of amino acid were measured and analysed. Otherwise change of testis and thyroid gland by hormone treatment were investigated histologically. The results obtained were as follows. 1. The effectst of caponization and hormone treatment upon meat production were; 1) Body weight of cockerels in D. E. S. group without caponization was increased. upon 96.86% than initial period and A. C. T. H. group was 104.22% but other groups and all carponization groups were lighter than those of control group. 2) Weekly body gain of D. E. S. group without caponization was best showing the significance (102.69 g) and the group with caponization were lower than those groups without caponization. 3) Carcass yield was best in Testo. group without caponization (831.2 g) and the group with caponization were lower than the group without caponization. 4) Carcass rate was highest in A. C. T. H. group with caponization and (67.22%) lowest in Testo. group without caponization (63.37%), but any significance was not recognized. 2. The effects of caponizatitn and hormone treatments upon the coposition of meat and amino acids were; 1) Any significance was not recognized between treated and untreated group about change of moisture, crude protein, crude ash and glycogen contents in meat. 2) Fat co tent in muscle in the all treated groups were higher than that of control group. 3) Extracts of group without caponization were higher than those of groups with caponization. 4) Lysin contents were highest in D. E. S. group with caponization (11. 12/ 16.0 g N) and generelly Testo. group was lower compared with D. E. S. group. 5) Histidine and Arginine contents were higher in the groups with caponization than without caponization. 6) Aspartic acid content were higher in D. E. S. group and A. C. T. H. group without depend on caponization. 7) Treonine content was higher in Testo. group without caponization and in the group with caponization and without hormone treatment compared with those of control group without caponization. 8) Serine content was decreased in the group with caponization and increased by D. E. S. and A. C. T. H treatment groups and glutamic acid was also decreased in Testo. group with out caponization. 9) Cystine content was decreased by Testo. treatment and was not appeared in Testo. group without caponization. 10) Valine content was lower in control group with caponization but significance was not recognized between other groups and control group without caponization. 11) Glycine, Alanine, Methionine. Isoleucine, Leucine, Thyrosine and Phenylalanine contents were not so difference between hormone treated groups and control group without caponization. 3. The effects of caponization and hormone treatment upon the change of organs were: 1) The weight of all organs were heaviest in D. E. S. group without caponization (18.5g) and lightest in A. C. T. H. group without caponization (155. 3g) but no significance was recognized between hormone treatment groups. 2) Heart weight was heaviest in D. E. S. group without caponization (7.46 g) and lightest in Testo. group without caponization (5.95 g). 3) Liver weight was heaviest in D. E. S. group without caponization(32.89g) and lightest in hormone untreated group with caponization(29.66g). Significance was not recognized. 4) Spleen weight was heaivest in Testo. group with caponization (3.22 g) and lightest in D. E. S. group without caponization(2.00g) in contrast with the other groups. High significance was recognized among the groups (P<0.01). 5) Cloacal thymus weight was lightest in D. E. S. group with or without caponization compared with control group without caponization. High significance was recognized among the groups. 6) Muscle fat content was not appeared in A. C. T. H. group with caponization, but it was highly increased in D. E. S. group with or without caponization. 7) Testis weight was lightest in D. E. S. group (0.38g) compared with control group (2.66g). Significance was recognized among the groups. 8) Large intestine, small intestine and cecum weight and length were heavier and longer in D. E. S. group without caponization and control group without caponization was lighter than those of hormone treated groups. 4. The effects of caponization and hormone treatment upon histological change of testis and thyroid gland: 1) The histological change of testis was significantly appeared in D. E. S. group that seminifirous tubles was slowly atrophied, the funtion of spernatogenesis was ceased, spermatocyte was changed as degeneration by pyknosis and karyorrhexis and interstitial cell was also atrophied, but in Testo. and A. C. T. H. group were similar as control group. 2) The histological change of thyroid gland in Testo. and A. C. T. H. groups without caponization were similar to that of control group without caponization, but in D. E. S. group without caponization, was changed squamously. Thyroid gland of the groups with caponization, epithelium of was atrophied and changed squamously as degeneration by pyknosis and karyorrhexis and the function of thyroid gland was slowly ceased in colloid and in hormone treated group with caponization.