Data_Sheet_1_Differential effects of areca nut, tobacco, and lime on oral squamous cell carcinoma progression.docx

BackgroundConsumption of tobacco and its associated products has been linked with a high incidence of oral cancer in the Indian subcontinent. The present study aims to assess the differential effects of areca nut, tobacco, and slaked lime on the pathogenesis and progression of oral cancer.
MethodologyExtracts of areca nut (ANE), tobacco leaf (TLE), and lime water (LWE) were prepared and characterized. Oral keratinocyte cells (BICR10) were treated with the prepared extracts alone and in combinations. Cell apoptosis, necrosis, nuclear aberrations, and transcript expression profiles were assessed to study the cancer pathogenesis.
ResultsAn increase in apoptosis and necrosis was observed in the cells treated with ANE and ANE+LWE, while a decrease in apoptosis was observed in cells treated with TLE. We also found a variation in the size of nuclei and an increase in the number of multinuclei in the cells treated with TLE and TLE+LWE. Out of 24 head and neck cancer-related genes, we found significant upregulation in seven genes in ANE, six in ANE+LWE, 13 in TLE, 20 in LWE, and 14 in TLE+LWE treated cells. On pathway enrichment analysis, there were alterations in the “pathways in cancer,” “focal adhesion,” and “amoebiasis.”
ConclusionAlthough areca nut has been regarded as a carcinogen, its carcinogenic potential is lesser than tobacco and lime. Moreover, adjunctive use of tobacco and lime along with areca nut may be an essential factor in the onset and progression of oral cancer.

背景 在印度次大陆地区，烟草及其相关制品的消费与口腔癌的高发病率密切相关。本研究旨在评估槟榔果（Areca nut）、烟草及熟石灰（Slaked lime）对口腔癌发病机制与疾病进展的差异化影响。
方法学 制备并表征了槟榔果提取物（Areca nut Extract，简称ANE）、烟草叶提取物（Tobacco Leaf Extract，简称TLE）及石灰水提取物（Lime Water Extract，简称LWE）。采用单一或联合配制的提取物处理人口腔角质形成细胞（BICR10）。通过检测细胞凋亡、细胞坏死、核异常及转录表达谱，以探究癌症发病机制。
结果 经ANE及ANE+LWE处理的细胞中，细胞凋亡与坏死水平均升高；而经TLE处理的细胞则呈现细胞凋亡水平降低。此外，经TLE及TLE+LWE处理的细胞出现细胞核大小异常，且多核细胞数量增加。在24个头颈部癌相关基因中，ANE处理组有7个基因显著上调，ANE+LWE处理组有6个，TLE处理组有13个，LWE处理组有20个，TLE+LWE处理组有14个。通路富集分析显示，"癌症通路"、"黏着斑"及"阿米巴病"相关通路发生了显著改变。
结论 尽管槟榔果已被认定为致癌物，但其致癌潜力弱于烟草与熟石灰。此外，槟榔果与烟草、熟石灰的联合使用，可能是口腔癌发生与进展的关键影响因素。