Overexpression of adenovirus-mediated p27kip1 lacking the Jab1-binding region enhances cytotoxicity and inhibits xenografted human cholangiocarcinoma growth.
Overexpression of adenovirus-mediated p27kip1 lacking the Jab1-binding region enhances cytotoxicity and inhibits xenografted human cholangiocarcinoma growth.
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发表时间：
2009-06
影响因子：
2
通讯作者：
Satoru Shiraso;Y. Katayose;Kuniharu Yamamoto;M. Mizuma;S. Yabuuchi;Akira Oda;T. Rikiyama;T. Onogawa;H. Yoshida;Hiroki Hayashi;H. Ohtsuka;F. Motoi;S. Egawa;Junya Kato;M. Unno
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作者：
Satoru Shiraso;Y. Katayose;Kuniharu Yamamoto;M. Mizuma;S. Yabuuchi;Akira Oda;T. Rikiyama;T. Onogawa;H. Yoshida;Hiroki Hayashi;H. Ohtsuka;F. Motoi;S. Egawa;Junya Kato;M. Unno
细胞周期蛋白依赖性激酶抑制剂（CDK 1）p27（kip 1）是细胞周期的负调节剂，具有抗肿瘤作用。在我们前期的研究中，表达野生型p27（kip 1）的重组腺病毒（Adp 27-wt）诱导了细胞周期阻滞和凋亡，证明了p27与p53一样是一种抑癌基因。另一种腺病毒载体表达突变型p27（kip 1）（Adp 27-mt），抑制降解的泛素-蛋白酶体系统，表现出增加的蛋白质稳定性，并引起更强的诱导凋亡。最近，发现与Jab 1（Jun活化结合蛋白1）结合的p27（kip 1）蛋白从细胞核易位到胞质溶胶中，然后被26 S蛋白酶体系统降解。核质易位的抑制增加了p27（kip 1）的蛋白质稳定性，并且缺失Jab 1结合区的p27（kip 1）（p27-jab-d）不易位且不降解。因此，制备了一种新的表达p27-jab-d的重组腺病毒（Adp 27-jab-d），其能够诱导更大的细胞毒性。Adp 27-jab-d在比Adp 27-wt低3.3倍（IC（50））的浓度下抑制人胆管癌细胞系（TFK-1）的体外生长。此外，在皮下组织中注射TFK-1细胞的异种移植严重联合免疫缺陷（SCID）小鼠模型中，在肿瘤建立后通过肿瘤内注射Adp 27-jab-d（每天一次，持续3天）进行治疗，比Adp 27-wt或Adp 27-mt更强烈地抑制肿瘤生长，甚至诱导肿瘤消退。然而，流式细胞术TUNEL检测显示细胞凋亡几乎没有增强。Adp 27-jab-d被认为不仅诱导细胞凋亡而且诱导坏死，这是由于Adp 27-jab-d的特异性作用。因此，通过抑制p27（kip 1）的移位来增强细胞毒性，缺乏Jab 1结合区的p27（kip 1）可能用于癌症治疗。控制蛋白定位不仅可能成为癌症治疗的新靶点，而且可能成为其他疾病的新靶点。
The cyclin-dependent kinase inhibitor (CDK1) p27(kip1) is a negative regulator of cell cycling and has antitumor effects. In our previous study, the recombinant adenovirus expressing wild-type p27(kip1) (Adp27-wt) induced cell cycle arrest and apoptosis, and proved that p27 is a tumor suppressor gene like p53. Another adenovirus vector expressing mutant p27(kip1) (Adp27-mt), which inhibited degradation by the ubiquitin-proteasome system, showed increased protein stability and caused a stronger induction of apoptosis. Recently, the p27(kip1) protein binding with Jab1 (Jun activating binding protein 1) was found to translocate from the nucleus into the cytosol, and then become degraded by the 26S proteasome system. The inhibition of nuclear-cytoplasmic translocation increases the protein stability of p27(kip1) and p27(kip1) with a deletion of the Jab1-binding region (p27-jab-d) is not translocated and not degraded. Therefore, a new recombinant adenovirus (Adp27-jab-d) expressing p27-jab-d was made which was able to induce greater cytotoxicity. Adp27-jab-d inhibited the growth of human cholangiocarcinoma cell line (TFK-1) cells in vitro at 3.3 times (IC(50)) lower concentration than Adp27-wt. Moreover, in a xenografted severe combined immuno-deficient (SCID) mouse model injected with TFK-1 cells in the subcutaneous tissue, treatment by intratumor injection of Adp27-jab-d once a day for 3 days after the tumor was established, inhibited tumor growth more strongly than Adp27-wt or Adp27-mt and even induced tumor regression. However, the flow cytometric TUNEL assay showed little enhancement of apoptosis. Adp27-jab-d was thought to induce not only apoptosis but also necrosis, which was due to a specific effect of the Adp27-jab-d. Thus, by enhancing the cytotoxicity through inhibiting the translocaton of p27(kip1), p27(kip1) lacking the Jab1-binding region might be useful for cancer therapy. The control protein localization might also be a new target not only for cancer treatment, but also other diseases.