Ji Xiaoqing, Ji Jiang, Shan Fang, Zhang Yongsheng, Chen Yongbing, Lu Xueguan
Department of Radiation Oncology, The Second Affiliated Hospital of Soochow University Suzhou, China.
Department of Pathology, The Second Affiliated Hospital of Soochow University Suzhou, China.
Int J Clin Exp Med. 2015 May 15;8(5):7002-8. eCollection 2015.
The purpose of present study was to investigate the radiosensitivity of lung cancer cells affected by human lung cancer-associated fibroblasts (CAFs) in transwell and direct co-culture in vitro. Human lung CAFs was obtained from fresh human lung adenocarcinoma tissue specimens by primary culture with tissue explants method, and was identified by immunofluorescence staining. The radiosensitivity of A549 and H1299 lung cancer cells in mono-culture, transwell and direct co-culture with human lung CAFs were investigated by clonogenic survival assay, respectively. Human lung CAFs were obtained successfully, and had specific immuno-staining with α-SMA, Vimentin, and FAP, and no cytokeratin-18 expression. The plating efficiency (PE) of A549 cells without receiving irradiation were 0.206±0.031, 0.210±0.007 (>0.05) and 0.352±0.023 (<0.05) in mono-culture, transwell and direct co-culture, respectively. The PE of H1299 cells were 0.200±0.039, 0.259±0.024 (>0.05) and 0.323±0.025 (<0.05) in mono-culture, transwell and direct co-culture, respectively. The clonogenic survival assay showed that the protection enhancement ratio of human lung CAFs in transwell and direct co-culture were 1.11 and 1.29 for A549 cells, and 1.15 and 1.25 for H1299 cells, respectively. The results suggested that human lung CAFs promote the radioresistance of lung cancer cells significantly. Its radioprotective effect may attribute to CAFs stimulating lung cancer cell proliferation.
本研究旨在探讨人肺癌相关成纤维细胞（CAFs）在体外Transwell小室共培养和直接共培养条件下对肺癌细胞放射敏感性的影响。采用组织块法原代培养新鲜人肺腺癌组织标本获取人肺CAFs，并通过免疫荧光染色进行鉴定。分别采用克隆形成存活试验研究A549和H1299肺癌细胞在单培养、Transwell小室共培养及与人肺CAFs直接共培养条件下的放射敏感性。成功获取人肺CAFs，其α -平滑肌肌动蛋白（α - SMA）、波形蛋白（Vimentin）和脂肪酸结合蛋白（FAP）呈特异性免疫染色，细胞角蛋白18（cytokeratin - 18）无表达。未接受照射的A549细胞在单培养、Transwell小室共培养及直接共培养条件下的接种效率（PE）分别为0.206±0.031、0.210±0.007（>0.05）和0.352±0.023（<0.05）。H1299细胞在单培养、Transwell小室共培养及直接共培养条件下的PE分别为0.200±0.039、0.259±0.024（>0.05）和0.323±0.025（<0.05）。克隆形成存活试验显示，Transwell小室共培养及直接共培养条件下人肺CAFs对A549细胞的保护增强率分别为1.11和1.29，对H1299细胞分别为1.15和1.25。结果表明，人肺CAFs可显著促进肺癌细胞的放射抗性。其辐射防护作用可能归因于CAFs刺激肺癌细胞增殖。
