CAJ | 학술논문

目的：构建肺癌放射抗性细胞系，观察细胞在形态、凋亡、侵袭迁移能力和上皮间质转化的改变。方法采用小剂量等分割照射法、亚致死剂量照射法及梯度递增照射法筛选肺癌A549和H1299放射抗性细胞系，显微镜下观察细胞形态学改变，克隆形成实验检测放射敏感性，CCK?8法检测细胞受照后存活率；流式细胞仪检测凋亡率；Transwell实验检测细胞侵袭迁移能力的变化；蛋白印迹法检测上皮间质转化标志蛋白表达。结果梯度递增照射法构建肺癌放射抗性细胞系可行性好，得到放射抗性细胞A549R和H1299R，形态学观察示放射抗性细胞向间质细胞形态转变。分别以A549和H1299细胞为对照，A549R和H1299R细胞D0、Dq、SF2增加（P＝0．017和0．033、0．001和0．000、0．000和0．008），α和α／β值减少（ P＝0．018和0．001、0．007和0．009）。 A549R和H1299R细胞在不同照射剂量下存活率均大于对照组（P值均＜0．05），受照后凋亡率降低（P＝0．02和0．01），侵袭率及迁徙率增高（ P＝0．000和0．001及0．001和0．002），E?钙黏蛋白表达下调和波形蛋白表达增高（ P＝0．00和0．01及P＝0．02和0．01）。结论成功构建肺癌放射抗性细胞系，其侵袭迁移能力增强，并且可能与上皮间质转化有关系。
목적：구건폐암방사항성세포계，관찰세포재형태、조망、침습천이능력화상피간질전화적개변。방법채용소제량등분할조사법、아치사제량조사법급제도체증조사법사선폐암A549화H1299방사항성세포계，현미경하관찰세포형태학개변，극륭형성실험검측방사민감성，CCK?8법검측세포수조후존활솔；류식세포의검측조망솔；Transwell실험검측세포침습천이능력적변화；단백인적법검측상피간질전화표지단백표체。결과제도체증조사법구건폐암방사항성세포계가행성호，득도방사항성세포A549R화H1299R，형태학관찰시방사항성세포향간질세포형태전변。분별이A549화H1299세포위대조，A549R화H1299R세포D0、Dq、SF2증가（P＝0．017화0．033、0．001화0．000、0．000화0．008），α화α／β치감소（ P＝0．018화0．001、0．007화0．009）。 A549R화H1299R세포재불동조사제량하존활솔균대우대조조（P치균＜0．05），수조후조망솔강저（P＝0．02화0．01），침습솔급천사솔증고（ P＝0．000화0．001급0．001화0．002），E?개점단백표체하조화파형단백표체증고（ P＝0．00화0．01급P＝0．02화0．01）。결론성공구건폐암방사항성세포계，기침습천이능력증강，병차가능여상피간질전화유관계。
Objective To establish radiation?resistant lung carcinoma cell lines, and to investigate the changes in morphology, apoptosis, invasive migration, and epithelial?mesenchymal transition ( EMT) in cells. Methods The radiation?resistant lung carcinoma cell lines were obtained by exposure of lung carcinoma cell lines, A549 and H1299, to radiation with a low dose in fractions, a sublethal dose, or a gradually increasing dose. The morphological changes in cells, radiosensitivity, survival rates after exposure, apoptosis rates, changes in invasive migration, and expression of EMT marker proteins were evaluated using microscopy, colony formation assay, CCK?8 assay, flow cytometry, transwell migration assay, and Western blot, respectively. Results Radiation with a gradually increasing dose successfully induced the radiation?resistant cell lines, A549R and H1299R. The morphological study showed that the morphology of radiation?resistant cells was converted to the morphology of mesenchymal cells. Compared with A549 and H1299 cells, the values of D0 , Dq , and SF2 were significantly increased in A549R ( P=0.017,P=0.001,P=0.000) and H1299R (P=0.033,P=0.000,P=0.008) cells, respectively;the values of α and α/β were significantly reduced in A549R (P=0.018;P=0.007) and H1299R (P=0.001;P=0.009) cells, respectively. The survival rates in A549R and H1299R cells after exposure to radiation with various doses were significantly higher than those in the control groups (all P<0.05). After exposure, the apoptosis rates were significantly reduced in A549R and H1299R cells ( P=0.02,P=0.01);the invasion and migration rates were significantly increased in A549R (P=0.000;P=0.001) and H1299R (P=0.001,P=0.002) cells;the expression of E?cadherin was significantly down?regulated in A549R and H1299R cells (P=0.00,P=0.01), while the expression of vimentin was significantly elevated in A549R and H1299R cells ( P= 0. 02, P= 0. 01 ) . Conclusions The radiation?resistant lung carcinoma cell lines are successfully established. Both cell lines show enhanced invasion and migration, which may be associated with EMT.