Kanzaki Hirotaka, Mukhopadhya Nishit K, Cui Xiaojiang, Ramanujan V Krishnan, Murali Ramachandran
1 Research Division of Immunology, Department of Biomedical Sciences, Cedars-Sinai Medical Center , Los Angeles, California.
5 Current address: Department of Pharmacy, Okayama University Hospital , Okayama, Japan .
Monoclon Antib Immunodiagn Immunother. 2016 Feb;35(1):1-11. doi: 10.1089/mab.2015.0056. Epub 2016 Jan 21.
A major clinical problem in the treatment of breast cancer is mortality due to metastasis. Understanding the molecular mechanisms associated with metastasis should aid in designing new therapeutic approaches for breast cancer. Trastuzumab is the main therapeutic option for HER2+ breast cancer patients; however, the molecular basis for trastuzumab resistance (TZR) and subsequent metastasis is not known. Earlier, we found expression of basal-like molecular markers in TZR tissues from patients with invasive breast cancer.(( 1 )) The basal-like phenotype is a particularly aggressive form of breast cancer. This observation suggests that TZR might contribute to an aggressive phenotype. To understand if resistance to TZR can lead to basal-like phenotype, we generated a trastuzumab-resistant human breast cancer cell line (BT-474-R) that maintained human epidermal growth factor receptor 2 (HER2) overexpression and HER2 mediated signaling. Analysis showed that nuclear factor-kappa B (NF-κB) was constitutively activated in the BT-474-R cells, a feature similar to the basal-like tumor phenotype. Pharmacologic inhibition of NF-κB improved sensitivity of BT-474-R cells to trastuzumab. Interestingly, activation of HER2 independent NF-κB is not shown in luminal B breast cancer cells. Our study suggests that by activating the NF-κB pathway, luminal B cells may acquire a HER2+ basal-like phenotype in which NF-κB is constitutively activated; this notion is consistent with the recently proposed "progression through grade" or "evolution of resistance" hypothesis. Furthermore, we identified IKK-α/IKK-β and nuclear accumulation of RelA/p65 as the major determinants in the resistant cells. Thus our study additionally suggests that the nuclear accumulation of p65 may be a useful marker for identifying metastasis-initiating tumor cells and targeting RelA/p65 may limit metastasis of breast and other cancers associated with NF-κB activation.
乳腺癌治疗中的一个主要临床问题是转移导致的死亡率。了解与转移相关的分子机制应有助于设计新的乳腺癌治疗方法。曲妥珠单抗是HER2阳性乳腺癌患者的主要治疗选择;然而,曲妥珠单抗耐药(TZR)及随后转移的分子基础尚不清楚。此前,我们在浸润性乳腺癌患者的TZR组织中发现了基底样分子标志物的表达。((1))基底样表型是一种特别侵袭性的乳腺癌形式。这一观察结果表明,TZR可能促成侵袭性表型。为了解对TZR的耐药是否会导致基底样表型,我们构建了一种曲妥珠单抗耐药的人乳腺癌细胞系(BT-474-R),该细胞系维持人表皮生长因子受体2(HER2)的过表达及HER2介导的信号传导。分析表明,核因子-κB(NF-κB)在BT-474-R细胞中持续激活,这一特征与基底样肿瘤表型相似。对NF-κB的药物抑制提高了BT-474-R细胞对曲妥珠单抗的敏感性。有趣的是,在腔面B型乳腺癌细胞中未显示HER2非依赖性NF-κB的激活。我们的研究表明,通过激活NF-κB途径,腔面B型细胞可能获得一种HER2阳性基底样表型,其中NF-κB持续激活;这一观点与最近提出的“逐级进展”或“耐药演变”假说一致。此外,我们确定IKK-α/IKK-β和RelA/p65的核积累是耐药细胞中的主要决定因素。因此,我们的研究还表明,p65的核积累可能是识别转移起始肿瘤细胞的有用标志物,靶向RelA/p65可能会限制乳腺癌和其他与NF-κB激活相关癌症的转移。