Antitumor effect of TAT-oxygen-dependent degradation-caspase-3 fusion protein specifically stabilized and activated in hypoxic tumor cells.
Antitumor effect of TAT-oxygen-dependent degradation-caspase-3 fusion protein specifically stabilized and activated in hypoxic tumor cells.
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TAT-氧依赖性降解-caspase-3融合蛋白的抗肿瘤作用在缺氧肿瘤细胞中特异性稳定和激活。
作者：
H. Harada;M. Hiraoka;S. Kizaka
人类实体瘤含有低氧区域，其具有比正常组织低得多的氧张力。这些赋予对放疗和抗癌化疗的抵抗力，以及增加肿瘤转移的倾向。为了开发对实体瘤高度特异性的潜在治疗性蛋白质药物，我们构建了在缺氧肿瘤细胞中选择性稳定的融合蛋白。缺氧诱导因子-1 α的一部分ODD结构域与β-半乳糖苷酶（β-Gal）融合形成的模型融合蛋白--氧依赖性降解（ODD）-β-半乳糖苷酶（β-Gal）在模拟缺氧条件下培养的细胞中表现出增加的稳定性。当ODD-β-Gal进一步融合到HIV-TAT蛋白转导结构域（达特（47-57））并腹膜内注射到荷瘤小鼠时，生物活性融合蛋白在实体瘤中特异性稳定，但在正常组织中几乎检测不到。此外，当野生型（WT）胱天蛋白酶-3（Casp 3（WT））或其催化失活突变体与TAT-ODD融合并腹膜内注射至荷瘤小鼠时，肿瘤的大小通过施用TAT-ODD-Casp 3（WT）而不是TAT-ODD-突变体Casp 3而减小。TAT-ODD-Casp 3（WT）对荷瘤小鼠没有引起任何明显的副作用，表明融合蛋白在缺氧肿瘤细胞中的特异性稳定和活化。这些结果表明，使用细胞毒性TAT-ODD融合蛋白的蛋白治疗与放疗和化疗的组合可能提供消灭实体瘤的新策略。
Human solid tumors contain hypoxic regions that have considerably lower oxygen tension than normal tissues. These impart resistance to radiotherapy and anticancer chemotherapy, as well as predisposing to increased tumor metastases. To develop a potentially therapeutic protein drug highly specific for solid tumors, we constructed fusion proteins selectively stabilized in hypoxic tumor cells. A model fusion protein, oxygen-dependent degradation (ODD)-beta-galactosidase (beta-Gal), composed of a part of the ODD domain of hypoxia-inducible factor-1alpha fused to beta-Gal, showed increased stability in cultured cells under a hypoxia-mimic condition. When ODD-beta-Gal was further fused to the HIV-TAT protein transduction domain (TAT(47-57)) and i.p. injected to a tumor-bearing mouse, the biologically active fusion protein was specifically stabilized in solid tumors but was hardly detected in the normal tissue. Furthermore, when wild-type (WT) caspase-3 (Casp3(WT)) or its catalytically inactive mutant was fused to TAT-ODD and i.p. injected to a tumor-bearing mouse, the size of tumors was reduced by the administration of TAT-ODD-Casp3(WT) but not by TAT-ODD-mutant Casp3. TAT-ODD-Casp3(WT) did not cause any obvious side effects on tumor-bearing mice, suggesting specific stabilization and activation of the fusion protein in the hypoxic tumor cells. These results suggest that the combination of protein therapy using a cytotoxic TAT-ODD fusion protein with radiotherapy and chemotherapy may provide a new strategy for annihilating solid tumors.
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DOI：
10.1073/pnas.080072497
发表时间：
2000-04-25
影响因子：
11.1
作者：
Sutter, CH;Laughner, E;Semenza, GL
通讯作者：
Semenza, GL
影响因子：
10.5
作者：
Woo, M;Hakem, R;Mak, TW
通讯作者：
Mak, TW
影响因子：
11.2
作者：
David Gius;S. Ezhevsky;M. Becker-Hapak;Hikaru Nagahara;Michael C. Wei;Steven F. Dowdy
通讯作者：
David Gius;S. Ezhevsky;M. Becker-Hapak;Hikaru Nagahara;Michael C. Wei;Steven F. Dowdy
DOI：
10.1073/pnas.95.15.8461
发表时间：
1998-07-21
影响因子：
11.1
作者：
Liu, XS;Li, P;Wang, XD
通讯作者：
Wang, XD
DOI：
10.1016/s0360-3016(98)00329-0
发表时间：
1998-11-01
影响因子：
7
作者：
Raleigh, JA;Chou, SC;Horsman, MR
通讯作者：
Horsman, MR