Lopes Floro Kylie, Gillman Rhys, Wankell Miriam, Dewdney Brittany, Chilkuri Madhavi, Shackelford Ashley, Kuma Leslie, Powers Marcus, Hebbard Lionel
Radiation Oncology, Townsville University Hospital, Townsville, QLD 4814, Australia.
Centre for Molecular Therapeutics, Department of Molecular Genetics, Australian Institute of Tropical Medicine and Health, College of Medicine and Dentistry, James Cook University, Townsville, QLD 4811, Australia.
Cancers (Basel). 2025 Apr 17;17(8):1346. doi: 10.3390/cancers17081346.
: Smokers are at an increased risk of developing mucosal head and neck squamous cell cancers (HNSCCs) and have a worse prognosis when treated. The cellular and molecular mechanisms underlying the latter has not been established. We therefore developed an in vitro model to investigate the effects of radiation and smoking on mucosal HNSCCs. FaDu hypopharyngeal cancer cells were subjected to daily fractionated radiation and cultured with and without cigarette-smoke-exposed media. : The cells were characterised using assays for tumour sphere formation, proliferation, migration, invasion, CD44 and ALDH expression, and next generation sequencing. We also evaluated CD44 and ALDH1 expression in patient tumour samples. : Radiation and smoking separately reduced FaDu tumour sphere/Cancer Stem Cell (CSC) number and proliferation, and increased cell migration and invasion. Combined, they further reduced CSC number proliferation and promoted migration. CD44 and ALDH co-expression was reduced in conditions with cigarette smoke. Through next generation sequencing, radiation and smoking produced a gene signature related to cell invasion, angiogenesis, and survival. Immunohistochemistry for CD44 and ALDH1 on patient tumour specimens did not demonstrate a relationship with smoking status, supported our in vitro findings. : The data show the utility of a new experimental model to test the combination of radiation and smoking on mucosal HNSCCs behaviour.
吸烟者患头颈部黏膜鳞状细胞癌（HNSCCs）的风险增加，且治疗时预后较差。后者潜在的细胞和分子机制尚未明确。因此，我们建立了一个体外模型来研究辐射和吸烟对黏膜HNSCCs的影响。将FaDu下咽癌细胞进行每日分次辐射，并在有或没有接触香烟烟雾的培养基中培养。通过肿瘤球形成、增殖、迁移、侵袭、CD44和ALDH表达检测以及下一代测序对细胞进行表征。我们还评估了患者肿瘤样本中CD44和ALDH1的表达。辐射和吸烟分别减少了FaDu肿瘤球/癌症干细胞（CSC）数量和增殖，并增加了细胞迁移和侵袭。两者联合作用时，进一步减少了CSC数量和增殖，并促进了迁移。在接触香烟烟雾的条件下，CD44和ALDH的共表达降低。通过下一代测序，辐射和吸烟产生了与细胞侵袭、血管生成和存活相关的基因特征。对患者肿瘤标本进行CD44和ALDH1免疫组织化学检测未发现与吸烟状态有关，这支持了我们的体外研究结果。这些数据表明了一种新的实验模型在测试辐射和吸烟联合作用对黏膜HNSCCs行为影响方面的实用性。
