Department of Otolaryngology‑Head and Neck Surgery, University of California at San Francisco, San Francisco, CA 94143, USA.
Department of Medicine‑Hematology/Oncology, University of Arizona, Tucson, AZ 85724, USA.
Mol Med Rep. 2019 Dec;20(6):5335-5344. doi: 10.3892/mmr.2019.10766. Epub 2019 Oct 23.
Patients who have undergone curative‑intent therapy for head and neck squamous cell carcinoma (HNSCC) exhibit a high rate of development of second primary tumors (SPTs), which are frequently lethal. A chemoprevention strategy that prevents SPTs would have a major impact on patient outcomes. Sulforaphane, a naturally‑occurring compound derived from cruciferous vegetables exhibits chemopreventive activity against HNSCC in a preclinical model. The effects of sulforaphane are considered to be mediated, in large part, through increased protein expression of the transcription factor nuclear factor erythroid 2‑related factor 2 (NRF2). Development of sulforaphane chemoprevention for HNSCC would benefit from the identification of robust biomarkers of sulforaphane activity in HNSCC cells and normal mucosal epithelial cells. The present study revealed that sulforaphane potently induces multiple oxidative stress‑associated genes at the RNA and protein levels, in HNSCC cells and Het‑1A cells, a non‑tumorigenic mucosal epithelial cell line. In the present analysis, HMOX1 and HSPA1A were identified as the most highly upregulated genes following sulforaphane treatment, suggesting their potential value as biomarkers to guide clinical trials. Sulforaphane induction of HMOX1 and HSPA1A was validated in vivo in murine tissues. Furthermore, the impact of sulforaphane treatment of HNSCC cells on the expression levels of natural killer group 2D (NKG2D) and DNAX accessory molecule‑1 (DNAM‑1) ligands, which are activators of natural killer (NK) cells, was examined. NRF2‑dependent upregulation of the NKG2D ligand MICA/B was observed. However, only one of the six HNSCC cell lines studied exhibited enhanced sensitivity to NK cell‑mediated killing following sulforaphane treatment, suggesting that this may not be a general mechanism of sulforaphane chemopreventive activity in HNSCC. In summary, the present study identified robust biomarkers of sulforaphane activity in HNSCC and normal tissues, supporting their application in the development of sulforaphane chemoprevention approaches for HNSCC.
接受头颈部鳞状细胞癌(HNSCC)根治性治疗的患者发生第二原发肿瘤(SPT)的风险很高,而 SPT 通常是致命的。如果有一种化学预防策略能够预防 SPT,将对患者的预后产生重大影响。在临床前模型中,从十字花科蔬菜中提取的天然化合物——萝卜硫素对 HNSCC 具有化学预防作用。萝卜硫素的作用被认为主要是通过增加转录因子核因子红细胞 2 相关因子 2(NRF2)的蛋白表达来介导的。开发用于预防 HNSCC 的萝卜硫素化学预防剂将受益于在 HNSCC 细胞和正常黏膜上皮细胞中鉴定出萝卜硫素活性的稳健生物标志物。本研究表明,萝卜硫素在 HNSCC 细胞和非致瘤性黏膜上皮细胞 Het-1A 中,在 RNA 和蛋白质水平上强烈诱导多种与氧化应激相关的基因。在本分析中,发现 HO-1 和 HSPA1A 是萝卜硫素处理后上调最明显的基因,表明它们具有作为指导临床试验的生物标志物的潜在价值。在小鼠组织中验证了萝卜硫素诱导 HO-1 和 HSPA1A 的作用。此外,还研究了萝卜硫素处理 HNSCC 细胞对自然杀伤细胞(NK)激活物 NKG2D 和 DNAX 辅助分子-1(DNAM-1)配体的表达水平的影响。观察到 NRF2 依赖性上调 NKG2D 配体 MICA/B。然而,在所研究的 6 种 HNSCC 细胞系中,只有一种细胞系在萝卜硫素处理后对 NK 细胞介导的杀伤表现出增强的敏感性,这表明这可能不是萝卜硫素预防 HNSCC 化学预防活性的一般机制。总之,本研究鉴定了 HNSCC 和正常组织中萝卜硫素活性的稳健生物标志物,支持将其应用于开发预防 HNSCC 的萝卜硫素化学预防方法。