Medical treatment research by the gene cluster (EPR-1, dCK, and TP) which guides antineoplastic drug susceptibility reinforcement

指导抗肿瘤药物敏感性强化的基因簇（EPR-1、dCK、TP）的医疗研究

• 批准号: 13470262
• 负责人: TANIGAWA Nobuhiko
• 金额: $ 9.86万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470262/
• 关键词: survivin; EPR-1; HT29; apoptosis; Adenovirus vector; retrovirus vector; dThdPase cDNA; dThdPase cDNA

EPR-1 cDNA, a potential survivin antisense, was ligated into a plasmid vector (pMTN2) with metallothionein promoter. Then HT29 human colon adenocarcinoma cells (with positive survivin expression) were transfected with the pMTN2-EPR-1 plasmid vector. Apoptosis was induced in HT29-AS33 and HT29-AS49 subolones with high EPR-1 expression in the presence of Zinc and furthermore sensitivity to Cis-DDP and 5-FU in vitro was increased. Then, on the basis of future clinical development, EPR-1 expressing adenovirus vector (ad.CMV-EPR-1) was made and the in vivo antitumor treatment was assessed on nude mouse HT29 xenografts. When the xenografts were 8mm or more, Ad.CMV-EPR1 and Ad.CMV-LacZ were injected to the tumor bearing mice once. In former group tumor formation was reduced to 1/4 comparing to the controls. This, suggested that the effect was due to HT29 cell line apoptosis induction. In addition, efficacy of CDDP or 5-FU combination with Ad.CMV-EPR-1 was enhanced, with reduction in tumor vol … More ume of 25% and 16% respectively (Eur J Cancer, in Press). Moreover, we have previously shown that transfected cells with plasmid vector expressing dThdPase(TP) cDNA were more sensitive to 5'-DFUR and 5-FU(Br J Cancer 75,1997). Therefore, at the time of this study, TPcDNA (1.5 kb) was transferred to EcoRI site of a retroviral vector, -pNV7 and transfected murine adenocarcinoma cell lines of MC38-TP and MC38-Neo were obtained. In Western blot and ELISA analysis the expression of TP was detected in MC38-TP, but not in the controls and the growth rate of both subclones was identical. As increased sensitivity to 5-FU,5'-DFUR and Capecitabine was significantly observed in vitro for the former group, in vivo experiment was also studied using subcutaneously inoculated MC38-TP mouse models which confirmed the enhancement of 5'-DFUR and Capecitabine chemotherapeutic effect in MC38-TP (data in submission). Such chemosensitivity related gene studies may be useful to provide effective improvement in current cancer chemotherapy. Less

将survivin反义基因EPR-1 cDNA克隆到含有金属硫蛋白启动子的质粒载体pMTN 2中。然后用pMTN 2-EPR-1质粒载体转染Survivin阳性表达的HT 29人结肠腺癌细胞。锌可诱导EPR-1高表达的HT 29-AS 33和HT 29-AS 49亚克隆细胞凋亡，并可提高细胞对顺铂和5-氟尿嘧啶的体外敏感性。然后，在未来临床开发的基础上，制备表达EPR-1的腺病毒载体（ad.CMV-EPR-1），并在裸鼠HT 29异种移植物上评估体内抗肿瘤治疗。当异种移植瘤直径大于等于8 mm时，将Ad. CMV-EPR 1和Ad.CMV-LacZ注射给荷瘤小鼠一次。与对照组相比，前者肿瘤形成减少至1/4。这表明，该作用是由于HT 29细胞系凋亡诱导。此外，CDDP或5-FU联合Ad.CMV-EPR-1的疗效增强，肿瘤体积减少， ...更多信息 分别为25%和16%（Eur J Cancer，出版中）。此外，我们先前已经表明，用表达dThdR（TP）cDNA的质粒载体转染的细胞对5 ′-DFUR和5-FU更敏感（Br J Cancer 75，1997）。因此，在本研究时，将TPcDNA（1.5kb）转移到逆转录病毒载体-pNV 7的EcoRI位点，并获得了转染的鼠腺癌细胞系MC 38-TP和MC 38-Neo。在Western blot和ELISA分析中，在MC 38-TP中检测到TP的表达，而在对照中未检测到，并且两个亚克隆的生长速率相同。由于在体外观察到前一组对5-FU、5 ′-DFUR和卡培他滨的敏感性显著增加，因此还使用皮下接种的MC 38-TP小鼠模型研究了体内实验，其证实了MC 38-TP中5 ′-DFUR和卡培他滨化疗效果的增强（提交的数据）。这些化疗敏感性相关基因的研究可能有助于提供有效的改善目前的癌症化疗。少

项目成果

H.Shinohara: "Expression of HER2 human gastric cancer cells directly correlates with antitumor activity of a recombinant disulfide-stabilized anti-HER2 immunotoxin"Journal of Surgical Research. 102. 169-177 (2002)

H.Shinohara：“HER2 人胃癌细胞的表达与重组二硫键稳定的抗 HER2 免疫毒素的抗肿瘤活性直接相关”《外科研究杂志》。

M.Iwamoto, N.Tanigawa: "Prognostic value of tumor-infiltrating dendritic cells expressing CD38 in human breast carcinomas"Int J Cancer. 104(1). 92-97 (2003)

M.Iwamoto、N.Tanikawa：“表达 CD38 的肿瘤浸润树突状细胞在人类乳腺癌中的预后价值”Int J Cancer。

T.Nohara: "Expression of cell-cycle regulator p27 is correlated to the prognosis and ER expression in breast carcinoma patients"Oncology. 60. 94-100 (2001)

T.Nohara：“细胞周期调节因子 p27 的表达与乳腺癌患者的预后和 ER 表达相关”。

H.Kawasaki: "Expression of survivin correlates with apoptosis, proliferation, and angiogenesis during human colorectal tumorigenesis"Cancer. 91(11). 2026-2032 (2001)

H.Kawasaki：“生存素的表达与人类结直肠肿瘤发生过程中的细胞凋亡、增殖和血管生成相关”癌症。

T.Yamamoto: "Down-regulation of survivin expression by induction of effector cell protease receptor-1 reduces tumor growth potential with increased sensitivity to anticancer agents in human colon cancer"Eur J Cancer. 38(17). 2316-2324 (2002)

T.Yamamoto：“通过诱导效应细胞蛋白酶受体 1 下调生存素表达可降低肿瘤生长潜力，同时增加人类结肠癌对抗癌药物的敏感性”Eur J Cancer。