Yang David T, Young Ken H, Kahl Brad S, Markovina Stephanie, Miyamoto Shigeki
Department of Pharmacology, University of Wisconsin School of Medicine and Public Health, Madison, USA.
Mol Cancer. 2008 May 19;7:40. doi: 10.1186/1476-4598-7-40.
The proteasome inhibitor bortezomib can inhibit activation of the transcription factor NF-kappaB, a mechanism implicated in its anti-neoplastic effects observed in mantle cell lymphoma (MCL). However, NF-kappaB can be activated through many distinct mechanisms, including proteasome independent pathways. While MCL cells have been shown to harbor constitutive NF-kappaB activity, what fraction of this activity in primary MCL samples is sensitive or resistant to inhibition by bortezomib remains unclear.
Proteasome activity in the EBV-negative MCL cell lines Jeko-1 and Rec-1 is inhibited by greater than 80% after exposure to 20 nM bortezomib for 4 hours. This treatment decreased NF-kappaB activity in Jeko-1 cells, but failed to do so in Rec-1 cells when assessed by electrophoretic mobility shift assay (EMSA). Concurrently, Rec-1 cells were more resistant to the cytotoxic effects of bortezomib than Jeko-1 cells. Consistent with a proteasome inhibitor resistant pathway of activation described in mouse B-lymphoma cells (WEHI231) and a breast carcinoma cell line (MDA-MB-468), the bortezomib-resistant NF-kappaB activity in Rec-1 cells is inhibited by calcium chelators, calmodulin inhibitors, and perillyl alcohol, a monoterpene capable of blocking L-type calcium channels. Importantly, the combination of perillyl alcohol and bortezomib is synergistic in eliciting Rec-1 cell cytotoxicity. The relevance of these results is illuminated by the additional finding that a considerable fraction of primary MCL samples (8 out of 10) displayed bortezomib-resistant constitutive NF-kappaB activity.
Our findings show that bortezomib-resistant NF-kappaB activity is frequently observed in MCL samples and suggest that this activity may be relevant to MCL biology as well as serve as a potential therapeutic target.
蛋白酶体抑制剂硼替佐米可抑制转录因子NF-κB的激活,这一机制与其在套细胞淋巴瘤(MCL)中观察到的抗肿瘤作用有关。然而,NF-κB可通过多种不同机制激活,包括蛋白酶体非依赖性途径。虽然MCL细胞已被证明具有组成性NF-κB活性,但原发性MCL样本中该活性的哪一部分对硼替佐米抑制敏感或耐药仍不清楚。
EBV阴性的MCL细胞系Jeko-1和Rec-1在暴露于20 nM硼替佐米4小时后,蛋白酶体活性被抑制超过80%。通过电泳迁移率变动分析(EMSA)评估,这种处理降低了Jeko-1细胞中的NF-κB活性,但在Rec-1细胞中未降低。同时,Rec-1细胞比Jeko-1细胞对硼替佐米的细胞毒性作用更具抗性。与在小鼠B淋巴瘤细胞(WEHI231)和乳腺癌细胞系(MDA-MB-468)中描述的蛋白酶体抑制剂耐药激活途径一致,Rec-1细胞中硼替佐米耐药的NF-κB活性被钙螯合剂、钙调蛋白抑制剂和紫苏醇(一种能够阻断L型钙通道的单萜)抑制。重要的是,紫苏醇和硼替佐米联合使用在诱导Rec-1细胞毒性方面具有协同作用。另外的发现表明相当一部分原发性MCL样本(10个中有8个)显示出硼替佐米耐药的组成性NF-κB活性,这说明了这些结果的相关性。
我们的研究结果表明,硼替佐米耐药的NF-κB活性在MCL样本中经常观察到,提示这种活性可能与MCL生物学相关,并可作为潜在的治疗靶点。