2000 Fiscal Year Annual Research Report

血管新生抑制および転移・浸潤抑制を目的とした卵巣癌遺伝子治療に関する基礎的研究

Research Project

Project/Area Number	11671645
Research Institution	Jichi Medical University
Principal Investigator	大和田倫孝自治医科大学, 医学部, 助教授 (40203955)
Co-Investigator(Kenkyū-buntansha)	小澤敬也自治医科大学, 医学部, 教授 (30137707) 小林浩浜松医科大学, 医学部, 講師 (40178330) 鈴木光明自治医科大学, 医学部, 教授 (50110870)
Keywords	卵巣癌 / AAVベクター / 遺伝子導入 / NK4 / UTI / thymidylate synthase
Research Abstract	Hepatocyte growth factor(HGF)のantagonistであるNK4,およびurinary trypsin inhibitor(UTI)には,卵巣癌の腹膜播種,浸潤抑制作用があることをすでに我々は確認している。本年度は卵巣癌遺伝子治療への臨床応用を目指し,NK4およびUTIのウイルスベクターの作製を試みた。非病原性で,非分裂細胞への導入が可能であるアデノ随伴ウイルス(AAV)への導入を行った。NK4発現ベクター(pCMV-NK4-IRES-bsr)およびUTI発現ベクター(pCMV-UTI)にITR配列を付加し,アデノフリーシステムによりNK4発現AAVベクター(AAV-NK4)およびUTI発現ベクター(AAV-UTI)の作製に成功した。これらのウイルスベクターを卵巣癌細胞株OVCAR-3を対象に,感染実験を施行した。その結果,MOI10^5〜10^6において培養上清中に,NK4およびUTIの分泌が確認され,またscratch wound assay,skim-milk assayによりそれぞれの活性も確認された。またUTIの活性基であるHI-8を取り出し,ATF-HI-8発現ベクター(pCMV-ATF-HI-8-IRES-bsr)作製に成功した。子宮頚癌細胞株SKG-IIにthymidylate synthase(TS)を遺伝子導入して作製されたTS強発現株(SKG-II/TS)は,コントロール(SKG-II/LUC)に比べて放射線感受性の低下が確認された。したがってTS高発現子宮頚癌症例の予後不良の原因のひとつに,放射線治療に対する抵抗性が示唆された。TS強発現とDNA修復能の関係をみるため,DNA修復関連物質であるXRCC-1,DNA polymerase β,DNA ligase IIIなどの活性を検討している。

Research Products
(10 results)

All Other

All Publications (10 results)

[Publications] Ohwada,M.: "Mutational analysis of transforming growth factor beta receptor type II and DNA mismatch repair genes in sporadic endometria carcinomas with microsatellite instability"Oncol Rep. 7. 789-792 (2000)
[Publications] Ohwada,M.: "DNA replication errors are frequent in mucinous cystadenocarcinoma of the ovary"Cancer Genet Cytogenet. 117. 61-65 (2000)
[Publications] Suzuki,M.: "DNA replication error is frequent in ovarian granulosa cell tumors"Cancer Genet Cytogenet. 122. 55-58 (2000)
[Publications] Suzuki,M.: "Lymph node metastasis in stage I epithelial ovarian cancer"Gynecol Oncol. 79. 305-308 (2000)
[Publications] Suzuki,M.: "Assessment of proliferation index with MIB-1 as a prognostic factor in radiation therapy for cervical cancer"Gynecol Oncol. 79. 300-304 (2000)
[Publications] Sugiyama,T.: "Clinical characteristics of clear cell carcinoma of the ovary : a distinc histologic type with poor prognosis and resistance to platinum-based chemotherapy"Cancer. 88. 2584-2589 (2000)
[Publications] Suzuki,M.: "Clinical significance of combined use of macrophage colony-stimulating factor and squamous cell carcinoma antigen as a selective diagnostic marker for squamous cell carcinoma arising in mature cystic teratoma of the ovary"Gynecol Oncol. 77. 405-409 (2000)
[Publications] Suzuki,M.: "Mutation of K-RAS protooncogene and loss of heterozygosity on 6q27 in serous and mucinous ovarian carcinomas"Cancer Genet Cytogenet. 118. 132-135 (2000)
[Publications] Saga,Y.,Suzuki,M.: "An in vitro examination of a 5-fluorouracil regimen involving continuous venous infusion using cultured cell lines derived from ovarian cancers"Oncol Rep. 118. 625-628 (2000)
[Publications] Kobayashi,H.: "Suppression of urokinase-type plasminogen activator expression from human ovarian cancer cells by urinary trypsin inhibitor"Biochim Biophys Acta. 1481. 310-316 (2000)

2000 Fiscal Year Annual Research Report

血管新生抑制および転移・浸潤抑制を目的とした卵巣癌遺伝子治療に関する基礎的研究

Principal Investigator

大和田 倫孝 自治医科大学, 医学部, 助教授 (40203955)

Research Products

[Publications] Ohwada,M.: "Mutational analysis of transforming growth factor beta receptor type II and DNA mismatch repair genes in sporadic endometria carcinomas with microsatellite instability"Oncol Rep. 7. 789-792 (2000)

[Publications] Ohwada,M.: "DNA replication errors are frequent in mucinous cystadenocarcinoma of the ovary"Cancer Genet Cytogenet. 117. 61-65 (2000)

[Publications] Suzuki,M.: "DNA replication error is frequent in ovarian granulosa cell tumors"Cancer Genet Cytogenet. 122. 55-58 (2000)

[Publications] Suzuki,M.: "Lymph node metastasis in stage I epithelial ovarian cancer"Gynecol Oncol. 79. 305-308 (2000)

[Publications] Suzuki,M.: "Assessment of proliferation index with MIB-1 as a prognostic factor in radiation therapy for cervical cancer"Gynecol Oncol. 79. 300-304 (2000)

[Publications] Sugiyama,T.: "Clinical characteristics of clear cell carcinoma of the ovary : a distinc histologic type with poor prognosis and resistance to platinum-based chemotherapy"Cancer. 88. 2584-2589 (2000)

[Publications] Suzuki,M.: "Clinical significance of combined use of macrophage colony-stimulating factor and squamous cell carcinoma antigen as a selective diagnostic marker for squamous cell carcinoma arising in mature cystic teratoma of the ovary"Gynecol Oncol. 77. 405-409 (2000)

[Publications] Suzuki,M.: "Mutation of K-RAS protooncogene and loss of heterozygosity on 6q27 in serous and mucinous ovarian carcinomas"Cancer Genet Cytogenet. 118. 132-135 (2000)

[Publications] Saga,Y.,Suzuki,M.: "An in vitro examination of a 5-fluorouracil regimen involving continuous venous infusion using cultured cell lines derived from ovarian cancers"Oncol Rep. 118. 625-628 (2000)

[Publications] Kobayashi,H.: "Suppression of urokinase-type plasminogen activator expression from human ovarian cancer cells by urinary trypsin inhibitor"Biochim Biophys Acta. 1481. 310-316 (2000)

大和田倫孝自治医科大学, 医学部, 助教授 (40203955)