Department of Biology, Lander College, Touro University, New York, NY, 11367, USA.
Biology and Anatomy, New York Medical College, 10595, Valhalla, NY, USA.
Respir Res. 2024 Aug 19;25(1):317. doi: 10.1186/s12931-024-02924-w.
Basal cells (BCs) are bronchial progenitor/stem cells that can regenerate injured airway that, in smokers, may undergo malignant transformation. As a model for early stages of lung carcinogenesis, we set out to characterize cytologically normal BC outgrowths from never-smokers and ever-smokers without cancers (controls), as well as from the normal epithelial "field" of ever-smokers with anatomically remote cancers, including lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC) (cases).
Primary BCs were cultured and expanded from endobronchial brushings taken remote from the site of clinical or visible lesions/tumors. Donor subgroups were tested for growth, morphology, and underlying molecular features by qRT-PCR, RNAseq, flow cytometry, immunofluorescence, and immunoblot.
(a) the BC population includes epithelial cell adhesion molecule (EpCAM) positive and negative cell subsets; (b) smoking reduced overall BC proliferation corresponding with a 2.6-fold reduction in the EpCAM/ITGA6 /CD24 stem cell fraction; (c) LUSC donor cells demonstrated up to 2.8-fold increase in dysmorphic BCs; and (d) cells procured from LUAD patients displayed increased proliferation and S-phase cell cycle fractions. These differences corresponded with: (i) disparate NOTCH1/NOTCH2 transcript expression and altered expression of potential downstream (ii) E-cadherin (CDH1), tumor protein-63 (TP63), secretoglobin family 1a member 1 (SCGB1A1), and Hairy/enhancer-of-split related with YRPW motif 1 (HEY1); and (iii) reduced EPCAM and increased NK2 homeobox-1 (NKX2-1) mRNA expression in LUAD donor BCs.
These and other findings demonstrate impacts of donor age, smoking, and lung cancer case-control status on BC phenotypic and molecular traits and may suggest Notch signaling pathway deregulation during early human lung cancer pathogenesis.
基底细胞（BCs）是支气管祖/干细胞，可以再生受损的气道，而在吸烟者中，气道可能会发生恶性转化。作为肺癌发生早期阶段的模型，我们着手对从不吸烟者和无癌症（对照组）的吸烟者以及解剖上远处有癌症（包括肺腺癌[LUAD]和鳞状细胞癌[LUSC]）的正常上皮“场”中正常的 BC 外生进行细胞特征描述。
从远离临床或可见病变/肿瘤部位的支气管刷取物中培养和扩增原发性 BC。通过 qRT-PCR、RNAseq、流式细胞术、免疫荧光和免疫印迹对供体亚组进行生长、形态和潜在分子特征的检测。
（a）BC 群体包括上皮细胞黏附分子（EpCAM）阳性和阴性细胞亚群；（b）吸烟减少了整体 BC 增殖，EpCAM/ITGA6/CD24 干细胞比例降低了 2.6 倍；（c）LUSC 供体细胞显示出多达 2.8 倍的畸形 BC 增加；（d）从 LUAD 患者中获取的细胞显示出增殖和 S 期细胞周期分数增加。这些差异与：（i）不同的 NOTCH1/NOTCH2 转录本表达和潜在下游（ii）E-钙黏蛋白（CDH1）、肿瘤蛋白 63（TP63）、分泌球蛋白家族 1a 成员 1（SCGB1A1）和 Hairy/enhancer-of-split 相关与 YRPW 基序 1（HEY1）的表达改变；以及（iii）LUAD 供体 BC 中 EPCAM 减少和 NK2 同源盒-1（NKX2-1）mRNA 表达增加有关。
这些发现和其他发现表明供体年龄、吸烟和肺癌病例对照状态对 BC 表型和分子特征的影响，并可能提示 Notch 信号通路在人类肺癌发病早期失调。
