Modulation of Pancreatic Cancer Chemoresistance by Inhibition of TAK1
Modulation of Pancreatic Cancer Chemoresistance by Inhibition of TAK1
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DOI：
10.1093/jnci/djr243
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发表时间：
2011-08-01
影响因子：
10.3
通讯作者：
Chiao, Paul J.
中科院分区：
文献类型：
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作者：
Melisi, Davide;Xia, Qianghua;Chiao, Paul J.
研究背景TGF-β激活的激酶1（TAK 1）是一种丝裂原激活的蛋白激酶激酶，通过激活核因子κ B（NF-κ B）和激活蛋白1（activator protein-1）而抑制细胞凋亡信号通路，从而促进化疗药物的耐药。然而，它是不知道，如果抑制TAK 1是有效的，在减少化疗药物对pancreaticcancer.Methods的耐药性在人胰腺癌细胞株AsPc-1，PANC-1，MDAPanc-28，其中TAK 1表达沉默的小发夹RNA的荧光素酶报告基因测定NF-κ B活性。在AsPc-1、PANC-1、MDAPanc-28和Colo 357 FG细胞中靶向TAK 1激酶活性，暴露于增加剂量的选择性小分子抑制剂LYTAK 1 24小时。为了测试LYTAK 1与化学治疗剂组合的作用，用递增剂量的奥沙利铂、SN-38或吉西他滨与LYTAK 1组合处理AsPc-1、PANC-1、MDAPanc-28细胞和对照细胞。口服LYTAK 1的体内活性在具有表达胰蛋白酶的AsPc-1胰腺癌细胞的原位裸鼠模型（n = 40，每组5只）中进行评价。通过非线性回归分析体外增殖结果的差异的统计学显著性;使用对数秩检验确定小鼠存活率的差异。结果AsPc-1和MDAPanc-28 TAK 1基因敲除细胞的NF-κ B B活性显著低于其各自的对照细胞（相对荧光素酶活性：AsPc-1，平均值= 0.18，95%置信区间[ CI] = 0.10至0.27;对照，平均值= 3.06，95% CI = 2.31至3.80; MDAPanc-28，平均值= 0.30，95%CI = 0.13至0.46;对照，平均值= 4.53，95%CI = 3.43至5.63;均P < .001）。TAK 1抑制剂LYTAK 1在AsPc-1、PANC-1、MDAPanc-28和Colo 357 FG细胞中具有有效的体外细胞毒性活性，IC 50在5至40 nM之间。与对照细胞相比，LYTAK 1还增强了化疗剂奥沙利铂、SN-38和吉西他滨在AsPc-1、PANC-1和MDAPanc-28细胞中的细胞毒性（P < .001）。在裸鼠中，口服LYTAK 1加吉西他滨在统计学上显著降低了肿瘤负荷（吉西他滨vs吉西他滨+LYTAK 1，P = .03）和延长生存期（中位生存期：吉西他滨，82天vs吉西他滨+LYTAK 1，122天;结论基因沉默或抑制TAK 1激酶活性是一种潜在的逆转胰腺癌内在耐药的治疗方法。J Natl Cancer Inst 2011; 103：1190-1204
Background TGF-beta-activated kinase-1 (TAK1), a mitogen-activated protein kinase kinase kinase, functions in the activation of nuclear factor kappa B (NF-kappa B) and activator protein-1, which can suppress proapoptotic signaling pathways and thus promote resistance to chemotherapeutic drugs. However, it is not known if inhibition of TAK1 is effective in reducing chemoresistance to therapeutic drugs against pancreatic cancer.Methods NF-kappa B activity was measured by luciferase reporter assay in human pancreatic cancer cell lines AsPc-1, PANC-1, and MDAPanc-28, in which TAK1 expression was silenced by small hairpin RNA. TAK1 kinase activity was targeted in AsPc-1, PANC-1, MDAPanc-28, and Colo357FG cells with exposure to increasing doses of a selective small-molecule inhibitor, LYTAK1, for 24 hours. To test the effect of LYTAK1 in combination with chemotherapeutic agents, AsPc-1, PANC-1, MDAPanc-28 cells, and control cells were treated with increasing doses of oxaliplatin, SN-38, or gemcitabine in combination with LYTAK1. In vivo activity of oral LYTAK1 was evaluated in an orthotopic nude mouse model (n = 40, 5 per group) with luciferase-expressing AsPc-1 pancreatic cancer cells. The results of in vitro proliferation were analyzed for statistical significance of differences by nonlinear regression analysis; differences in mouse survival were determined using a log-rank test. All statistical tests were two-sided.Results AsPc-1 and MDAPanc-28 TAK1 knockdown cells had a statistically significantly lower NF-kappa B activity than did their respective control cell lines (relative luciferase activity: AsPc-1, mean = 0.18, 95% confidence interval [ CI] = 0.10 to 0.27; control, mean = 3.06, 95% CI = 2.31 to 3.80; MDAPanc-28, mean = 0.30, 95% CI = 0.13 to 0.46; control, mean = 4.53, 95% CI = 3.43 to 5.63; both P < .001). TAK1 inhibitor LYTAK1 had potent in vitro cytotoxic activity in AsPc-1, PANC-1, MDAPanc-28, and Colo357FG cells, with IC50 between 5 and 40 nM. LYTAK1 also potentiated the cytotoxicity of chemotherapeutic agents oxaliplatin, SN-38, and gemcitabine in AsPc-1, PANC-1, and MDAPanc-28 cells compared with control cells (P < .001). In nude mice, oral administration of LYTAK1 plus gemcitabine statistically significantly reduced tumor burden (gemcitabine vs gemcitabine plus LYTAK1, P = .03) and prolonged survival duration (median survival: gemcitabine, 82 days vs gemcitabine plus LYTAK1, 122 days; hazard ratio = 0.334, 95% CI = 0.027 to 0.826, P = .029).Conclusions The results of this study suggest that genetic silencing or inhibition of TAK1 kinase activity in vivo is a potential therapeutic approach to reversal of the intrinsic chemoresistance of pancreatic cancer. J Natl Cancer Inst 2011; 103: 1190-1204