Proteasome inhibition sensitizes non-small-cell lung cancer to gemcitabine-induced apoptosis
Proteasome inhibition sensitizes non-small-cell lung cancer to gemcitabine-induced apoptosis
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DOI：
10.1016/j.athoracsur.2004.04.029
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发表时间：
2004-10-01
影响因子：
4.6
通讯作者：
Jones, DR
中科院分区：
文献类型：
--
作者：
Denlinger, CE;Rundall, BK;Jones, DR
背景。我和我的同事之前已经证明，化疗激活非小细胞肺癌（NSCLC）的抗凋亡转录因子核因子(NF)- κ b。我们假设使用蛋白酶体抑制剂硼替佐米（Velcade）抑制NF-kappaB会使NSCLC对吉西他滨诱导的细胞凋亡敏感。致瘤性NSCLC细胞系（H157和A549）不使用吉西他滨、硼替佐米或两种化合物治疗。NF-kappaB的活性是通过核p65蛋白水平、电泳迁移率转移试验和NF-kappaB调控基因白介素-8、c-IAP2和Bcl-xL的逆转录聚合酶链反应来确定的。在同样处理的细胞中测定p21和p53蛋白水平。通过荧光活化细胞分选分析评估细胞周期失调。通过克隆实验、caspase-3激活和DNA片段化来定量细胞死亡和凋亡。如前所述，生成和处理NSCLC A549异种移植物。在4周的治疗期间评估肿瘤生长情况。采用方差分析进行统计学分析。吉西他滨增强核p65水平，NF-kappaB与DNA结合，以及所有NF-kappaB调节基因的转录。硼替佐米抑制了这些作用。吉西他滨和硼替佐米联合使用可增强p21和p53的表达，并分别诱导s期和G(2)/M细胞周期阻滞。通过caspase-3激活（p = 0.05）和DNA断裂（p = 0.02）测定，联合治疗可杀死80%的NSCLC细胞并诱导凋亡。联合治疗的非小细胞肺癌异种移植物生长明显慢于单独使用吉西他滨治疗的异种移植物（p = 0.02）。硼替佐米抑制吉西他滨诱导的NF-kappaB活化，使NSCLC在体内和体外致敏死亡。鉴于目前两种药物的临床可用性，这种联合治疗策略值得进一步研究，并可能代表一种合理的非小细胞肺癌患者的治疗策略。(C) 2004年由胸外科学会出版。
Background. My colleagues and I have previouslyshown that chemotherapy activates the antiapoptotic transcription factor nuclear factor (NF)-kappaB in non-small-cell lung cancer (NSCLC). We hypothesized that inhibition of NF-kappaB by using the proteasome inhibitor bortezomib (Velcade) would sensitize NSCLC to gemcitabine-induced apoptosis.Methods. Tumorigenic NSCLC cell lines (H157 and A549) were treated with nothing, gemcitabine, bortezomib, or both compounds. NF-kappaB activity was determined by nuclear p65 protein levels, electrophoretic mobility shift assays, and reverse transcription-polymerase chain reaction of the NF-kappaB-regulated genes interleukin-8, c-IAP2, and Bcl-xL. The p21 and p53 protein levels were determined in similarly treated cells. Cell-cycle dysregulation was assessed by fluorescence-activated cell sorting analysis. Cell death and apoptosis were quantified by clonogenic assays, caspase-3 activation, and DNA fragmentation. NSCLC A549 xenografts were generated and treated as noted previously. Tumor growth was assessed over a 4-week treatment period. Statistical analysis was performed with analysis of variance.Results. Gemcitabine enhanced nuclear p65 levels, NF-kappaB binding to DNA, and transcription of all NF-kappaB-regulated genes. Bortezomib inhibited each of these effects. Combined gemcitabine and bortezomib enhanced p21 and p53 expression and induced S-phase and G(2)/M cell-cycle arrests, respectively. Combined treatment killed 80% of the NSCLC cells and induced apoptosis, as determined by caspase-3 activation (p = 0.05) and DNA fragmentation (p = 0.02). NSCLC xenografts treated with combination therapy grew significantly slower than xenografts treated with gemcitabine alone (p = 0.02).Conclusions. Bortezomib inhibits gemcitabine-induced activation of NF-kappaB and sensitizes NSCLC to death in vitro and in vivo. This combined treatment strategy warrants further investigation and may represent a reasonable treatment strategy for select patients with NSCLC given the current clinical availability of both drugs. (C) 2004 by The Society of Thoracic Surgeons.