云锡矿粉离体诱导永生化大鼠肺泡Ⅱ型上皮细胞RLE-6TN恶性转化  

Study on the malignant transformation of immortalized rat alveolar cells type Ⅱ RLE-6TN induced by Yunnan tin mine dust

作　　者：孟庆印[1] 阮永华[2] 金克炜[2] 唐莹[2] 高倩[2] 边莉[3] 

机构地区：[1]昆明医学院附属肿瘤医院病理科,云南昆明650106 [2]昆明医学院病理教研室,650500 [3]昆明医学院第一附属医院病理科,650032

出　　处：《中国职业医学》2011年第4期276-279,282,共5页China Occupational Medicine

基　　金：国家自然科学基金项目(81060189);云南省应用基础研究基金项目(2010ZC107)

摘　　要：目的建立本实验云锡矿粉(无氡)诱导永生化大鼠肺泡Ⅱ型上皮细胞RLE-6TN恶性转化模型,观察在诱导过程中不同阶段的细胞形态、超微结构和某些相关蛋白改变。方法高、低2个剂量(200和50 mg/L)矿粉对RLE-6TN细胞进行72 h染毒,隔代染毒至第10代,常规传代直至软琼脂克隆形成实验阳性;通过透射电镜、流式细胞术、免疫印迹技术及免疫细胞化学观察实验细胞的改变。结果染毒24 h细胞内见大量矿粉,板层小体变性减少,出现细胞凋亡;第25代高剂量组及第30代高低剂量组的细胞均可在软琼脂上形成细胞集落,空白对照组克隆形成实验阴性(P<0.01);第30代高剂量组细胞增殖指数(PI)高于空白对照组;高剂量组转化细胞p53(+)、MDM2(+)、FHIT(-);空白对照组p53(-)、MDM2(+)、FHIT(+)。结论云锡矿粉可引起离体培养的RLE-6TN细胞发生恶性转化。p53阳性表达及FHIT失表达可作为RLE-6TN细胞转化的参考指标。Objective To set up a malignant transformation model of immortalized rat alveolar cells type Ⅱ（RLE-6TN）induced by Yunnan tin mineral dust（non radon）in vitro,to observe the changes of morphology,ultrastructure and the related proteins.Methods RLE-6TN cells were treated with tin mine dusts at the concentrations of 200 and 50 mg/L for 72 hours on every other generations for 10 generations,then the cells were transmitted routinely until the malignant transformation phenotype of cells were identified through observing the anchorage independent growth.The changes of RLE-6TN cells were observed by electron microscopy（EM）,flow cytometry（FCM）,immunocytochemistry（ICC）and western blot（WB）.Results 24 hours after the reaction of tin mine dust,a lot of tin mine dust inside the cells,decreased and degenerated osmiophilic lamellar bodies,and apoptosis were found in the RLE-6TN cells.At the 25th generation of high concentration group and at the 30th generation of low concentration group,intensive cell colonies in soft agar were found,showing a significant difference with the negative control（P0.01）.The cell proliferation index（PI）of the high concentration group was significantly higher than that of the control group.The results of ICC were p53（＋）、MDM2（＋）、FHIT（-）in high concentration group cells and p53（-）、MDM2（＋）、FHIT（＋）in the control group.Conclusion Yunnan tin mine dust can induce malignant transformation of RLE-6TN cells in vitro.Expression of p53 and lost expression of FHIT could be used as reference targets of malignant transformation of RLE-6TN cells.

关 键 词：肺泡Ⅱ型上皮 云锡矿粉 恶性转化 

分 类 号：R730.2[医药卫生—肿瘤] R135.2[医药卫生—临床医学]