Masciale Valentina, Grisendi Giulia, Banchelli Federico, D'Amico Roberto, Maiorana Antonino, Sighinolfi Pamela, Stefani Alessandro, Morandi Uliano, Dominici Massimo, Aramini Beatrice
Division of Thoracic Surgery, Department of Medical and Surgical Sciences for Children & Adults, University of Modena and Reggio Emilia, Modena, Italy.
Division of Oncology, Department of Medical and Surgical Sciences for Children & Adults, University of Modena and Reggio Emilia, Modena, Italy.
Front Oncol. 2019 Dec 18;9:1394. doi: 10.3389/fonc.2019.01394. eCollection 2019.
Lung cancer stem cells (CSCs) share many characteristics with normal stem cells, such as self-renewal and multipotentiality. High expression of aldehyde dehydrogenase (ALDH) has been detected in many tumors, particularly in the CSC compartment, and it plays an important role in tumor proliferation, metastasis, and drug resistance. CD44 is commonly used as a cell surface marker of cancer stem-like cells in epithelial tumors. The aim of this study was to isolate and analyze cancer stem-like cells from surgically removed specimens to compare lung adenocarcinoma (ADENO) and squamous (SQUAMO) cell carcinoma. The ALDEFLUOR assay was used to identify and sort ALDH and ALDH human lung cancer cells following tissue digestion. Fluorescence-activated cell sorting analysis for CD44 was performed with tumor cells. Quantitative real-time PCR was performed to assess the expression of SOX2 and NANOG as stemness markers. ALDH1A1 expression was additionally determined by immunohistochemistry. Anchorage-independent ALDH cell growth was also evaluated. ALDH ADENO and SQUAMO cells were cultured to analyze spheroid formation. All specimens contained 0.5-12.5% ALDH cells with 3.8-18.9% CD44-positive cells. SOX2 and NANOG relative expression in ALDH compared to ALDH cells in ADENO and SQUAMO was analyzed and compared between the histotypes. Immunohistochemistry confirmed the presence of ALDH1A1 in the sections. SOX2 and NANOG were expressed at higher levels in the ALDH subpopulation than in the ALDH subpopulation only in ADENO cells, and the opposite result was seen in SQUAMO cells. functional assays demonstrated that ALDH cells exhibited migration capacity with distinct behaviors between ALDH spheres in ADENO vs. SQUAMO samples. Our results highlight the importance of a better characterization of cancer stem-like cells in ADENO and SQUAMO histotypes. This may suggest new differential approaches for prognostic and therapeutic purposes in patients with non-small-cell lung cancer.
肺癌干细胞（CSCs）与正常干细胞具有许多共同特征，如自我更新和多能性。在许多肿瘤中，尤其是在癌症干细胞部分，已检测到醛脱氢酶（ALDH）的高表达，它在肿瘤增殖、转移和耐药性中起重要作用。CD44通常用作上皮肿瘤中癌症干细胞样细胞的细胞表面标志物。本研究的目的是从手术切除的标本中分离和分析癌症干细胞样细胞，以比较肺腺癌（ADENO）和鳞状细胞癌（SQUAMO）。采用ALDEFLUOR检测法在组织消化后鉴定和分选ALDH阳性和ALDH阴性人肺癌细胞。对肿瘤细胞进行CD44的荧光激活细胞分选分析。进行定量实时PCR以评估SOX2和NANOG作为干性标志物的表达。另外通过免疫组织化学测定ALDH1A1的表达。还评估了不依赖贴壁的ALDH细胞生长。培养ALDH阳性的ADENO和SQUAMO细胞以分析球体形成。所有标本含有0.5 - 12.5%的ALDH阳性细胞和3.8 - 18.9%的CD44阳性细胞。分析并比较了ADENO和SQUAMO中ALDH阳性细胞与ALDH阴性细胞相比SOX2和NANOG的相对表达。免疫组织化学证实切片中存在ALDH1A1。仅在ADENO细胞中，SOX2和NANOG在ALDH阳性亚群中的表达高于ALDH阴性亚群，而在SQUAMO细胞中则观察到相反的结果。功能测定表明，ALDH阳性细胞表现出迁移能力，ADENO与SQUAMO样本中ALDH阳性球体之间存在不同行为。我们的结果强调了更好地表征ADENO和SQUAMO组织学类型中癌症干细胞样细胞的重要性。这可能为非小细胞肺癌患者的预后和治疗目的提示新的鉴别方法。
