The basic study of the metabolic enzyme for prodrug gene therapy, and its application to gynecological malignant tumors
前药基因治疗代谢酶的基础研究及其在妇科恶性肿瘤中的应用
基本信息
- 批准号:11671609
- 负责人:
- 金额:$ 2.18万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:1999
- 资助国家:日本
- 起止时间:1999 至 2000
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
We had studied the basic research at the view points for prodrug gene therapy. Among the anti-cancer drugs for gynecologic tumors, now we have studied CPT-11 (as a pro-drug), carboxylesterase (as an activator for CPT-11), and the cancer cells' up-sensitibity to prodrug CPT-11 through the insersion of this enzyme's molecular gene.By Potter et al. reported that enzyme activity of Rabbit carboxylhydrate was seven times superior to Human carboxylhydrate. Using the RT-PCR methods, we had picked up the gene clone of 1.7 KB Rabbit carboxylesterase c-DNA from Rabbit liver, and also done the gene clone of 1.7 KB Human carboxylesterase c-DNA from human leukemia cell HEL.There after, to compare the activities among these enzymes, we made three chimeras by some combining those cDNAs, and put those into the pCMV-tag2a mammalian expression vectors that had frag-tag sequence at the N-terminals. Five kinds of expression vectors were introduced into gynecological tumor cells. Transiently every vectors were introduced into 293T cell line. After forty hours, every carboxylesterase activities were determined. Introduced two cell lines showed higher enzyme activity.At last, using Tet-off system, Rabbit carboxylesterase c-DNA inserted into expression vector, was introduced into ovarian cancer cell line SKOV3, and established two cell lines of hishly expression carboxylhydrate.
从前体药物基因治疗的角度对基础研究进行了综述。在妇科肿瘤的抗癌药物中,我们已经研究了CPT-11(作为前药)、羧酸酯酶(作为CPT-11的激活剂),以及通过插入该酶的分子基因使癌细胞对前药CPT-11增敏,Potter等报道兔羧化物的酶活性是人羧化物的上级7倍。本研究采用RT-PCR方法,从兔肝中克隆了1.7kB兔羧酸酯酶c-DNA,从人白血病细胞HEL中克隆了1.7kB人羧酸酯酶c-DNA,并将这两种酶的cDNA进行嵌合,并将其插入到N端带有片段标签序列的pCMV-tag 2a哺乳动物表达载体中。将5种表达载体导入妇科肿瘤细胞。将每种载体瞬时导入293 T细胞系。40 h后测定各羧酸酯酶活性。最后,利用Tet-off系统,将兔羧酸酯酶c-DNA插入表达载体,导入卵巢癌细胞株SKOV 3,建立了两株高表达羧酸酯酶的细胞株。
项目成果
期刊论文数量(9)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Mari Yotsu-Yamashita, et al: "Binding Properties of ^3H-PbTx-3 and ^3H-Saxitoxin to Brain Membranes and to Skeletal Muscle Membranes of Puffer Fish Fugu pardalies and the Primary Struture of a Voltage-Gated Na^+Channel α-Subunit (fMNal) from Skeletal Musc
Mari Yotsu-Yamashita 等人:“^3H-PbTx-3 和 ^3H-石房蛤毒素与河豚河豚鱼脑膜和骨骼肌膜的结合特性以及电压门控 Na^+通道 α 的主要结构-来自骨骼肌的亚基(fMNal)
- DOI:
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- 影响因子:0
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Julia A.Elvin, et al: "Paracrine Actions Of Grouth Diffrentiation Factor-9 in the Mammalian Ovary"Mol Endo. 13(6). 1035-1048 (1999)
Julia A.Elvin 等人:“哺乳动物卵巢中生长分化因子 9 的旁分泌作用”Mol Endo。
- DOI:
- 发表时间:
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- 影响因子:0
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Mari Yotsu-Yamashita, et al: "Binding Properties of ^3H-PbTx-3 and ^3H-Saxitoxin to Brain Membranes and to Skeletal Muscle Membranes of Puffer Fish Fugu pardalies and the Primary Struture of a Voltage-Gated Na^+Channel α -Subunit (fMNa1) from Skeletal Mus
Mari Yotsu-Yamashita 等人:“^3H-PbTx-3 和 ^3H-石房蛤毒素与河豚河豚鱼脑膜和骨骼肌膜的结合特性以及电压门控 Na^+通道 α 的主要结构-来自骨骼肌的亚基 (fMNa1)
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- 影响因子:0
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Akihiro Nawa, et al: "Prognostic factors of patients with yolk sac tumors of ovary"Am J Obstet Gynecol. 184. 1182-1188 (2001)
Akihiro Nawa 等人:“卵巢卵黄囊肿瘤患者的预后因素”Am J Obstet Gynecol。
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- 影响因子:0
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ANTHONY L.LAU, et al: "Acivin βC and βE Genes Are Not Essential for Mouse Liver Growth, Differentiation, and Regeneration"Mol Cell Biol. 20(16). 6127-6137 (2000)
ANTHONY L.LAU 等人:“Acivin βC 和 βE 基因对于小鼠肝脏生长、分化和再生不是必需的”Mol Cell Biol. 20(16)。
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{{ truncateString('GOTO Setsuko', 18)}}的其他基金
effect of the inhibitor to membrane-bound enzyme on the growth of invasion of cancer cells
膜结合酶抑制剂对癌细胞侵袭生长的影响
- 批准号:
06671642 - 财政年份:1994
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Induction of NHC antigens of trophoblastic cells using some cytokines and its application to the immunotherapy in choriocarcinoma
某些细胞因子诱导滋养层细胞NHC抗原及其在绒毛膜癌免疫治疗中的应用
- 批准号:
03670778 - 财政年份:1991
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)