Enzastaurin shows preclinical antitumor activity against human transitional cell carcinoma and enhances the activity of gemcitabine
Enzastaurin shows preclinical antitumor activity against human transitional cell carcinoma and enhances the activity of gemcitabine
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DOI：
10.1158/1535-7163.mct-09-0141
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发表时间：
2009-07-01
影响因子：
5.7
通讯作者：
Sonpavde, Guru
中科院分区：
文献类型：
--
作者：
Jian, Weiguo;Yamashita, Hideyuki;Sonpavde, Guru
恩扎洛宁是一种口服丝氨酸/苏氨酸激酶抑制剂，通过蛋白激酶C（PKC）-β和磷脂酰肌醇3-激酶/AKT途径抑制信号传导。我们在临床前评估了恩扎鲁肽单药治疗和恩扎鲁肽与吉西他滨联合治疗移行细胞癌（TCC）的疗效。对来自微阵列的105个人类样本进行免疫组织化学（IHC）以显示PKC-β的表达。测定了恩杂鲁肽和吉西他滨作为单药和联合用药对侵袭性人细胞系（p53和5637）和携带5637细胞的小鼠皮下异种移植物的临床前抗肿瘤活性。在体外对肿瘤细胞进行蛋白质印迹以检测通过PKC-β、GSK-3 β和AKT的信号传导。在体外测定对细胞迁移的影响。通过IHC测定体内增殖（Ki-67）、凋亡（裂解的半胱天冬酶-3）和血管生成（CD 31）的调节。对来自微阵列的人类TCC样本进行的IHC显示33%的肿瘤中有PKC-β的表达。在体外低微摩尔浓度下，Enzaelatin诱导显著的细胞凋亡并抑制增殖。通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四唑溴化铵试验，恩扎鲁肽和吉西他滨联合给药的体外抑制活性似乎具有协同作用。蛋白质印迹显示Akt、PKC-β和GSK-3 β磷酸化下调。Enzaelatin在较早的时间点抑制迁移，与抗增殖活性无关。与未处理的对照组相比，联合治疗在小鼠异种移植物中具有显著的上级抗肿瘤活性，而单一药物则没有。IHC显示，与对照组相比，联合治疗组Ki-67和CD 31降低，裂解的caspase-3增加。恩扎利汀对人TCC显示出临床前抗肿瘤活性，并增强吉西他滨的活性。[Mol癌症治疗2009;8（7）：1772-8]
Enzastaurin, an oral serine/threonine kinase inhibitor, suppresses signaling through protein kinase C (PKC)-beta and the phosphatidylinositol 3-kinase/AKT pathways. We preclinically evaluated enzastaurin alone and in combination with gemcitabine for transitional cell cancer (TCC). Immunohistochemistry (IHC) was done on 105 human samples from a microarray to show the expression of PKC-beta. The preclinical antitumor activity of enzastaurin and gemcitabine as single agents and in combination against aggressive human -lines (-SUP and 5637) and murine subcutaneous xenografts bearing 5637 cells was determined. Western Blot was done on tumor cells in vitro to detect signaling through PKC-beta, GSK-3 beta, and AKT. The effect on cell migration was determined in vitro. Modulation of proliferation (Ki-67), apoptosis (cleaved caspase-3), and angiogenesis (CD31) in vivo was determined by IHC. IHC done on human TCC samples from a microarray showed the expression of PKC-beta in 33% of tumors. Enzastaurin induced significant apoptosis and inhibited proliferation in vitro at low micromolar concentrations. The in vitro inhibitory activity of combination enzastaurin and gemcitabine by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay seemed synergistic. Western Blotting revealed down-regulation of Akt, PKC-beta, and GSK-3 beta phosphorylation. Enzastaurin inhibited migration at an earlier time point independent of antiproliferative activity. Combination therapy had significantly superior antitumor activity in murine xenografts compared with untreated controls, whereas single agents did not. IHC showed reduced Ki-67 and CD31 and increased cleaved caspase-3 with combination therapy compared with controls. Enzastaurin showed preclinical antitumor activity against human TCC and enhanced the activity of gemcitabine. [Mol Cancer Ther 2009;8(7):1772-8]