Vimentin,CTGF和AQP5在烟尘诱导肺泡Ⅱ型上皮细胞恶性转化模型中的表达  

The Expression of Vimentin,CTGF and AQP5 in the Model of Yunnan Tin Mine Induced Malignant Transformation of Lung Alveolar Epithelial Type Ⅱ Cells in Vitro

作　　者：唐莹[1] 徐天勇 边莉[3] 孟庆印[4] 金克炜[5] 阮永华[5] 

机构地区：[1]昆明医科大学电子显微镜室,云南昆明650500 [2]昆明医科大学科研实验中心,云南昆明650500 [3]昆明医科大学第一附属医院病理科,云南昆明650031 [4]昆明医科大学附属肿瘤医院病理科,云南昆明650032 [5]昆明医科大学基础医学院,云南昆明650500

出　　处：《昆明医科大学学报》2015年第8期1-5,共5页Journal of Kunming Medical University

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

摘　　要：目的观察波形蛋白(Vimentin)、结缔组织生长因子(conective tissue growth factor,CTGF)和水通道蛋白5(Aquaporin5,AQP5)在烟尘诱导肺泡Ⅱ型上皮细胞(ATⅡ)恶性转化模型中的表达.方法以永生化ATⅡ(RLE-6TN细胞株)为实验对象,分为高剂量染毒组、低剂量染毒组和空白对照组.高、低剂量染毒组分别使用终浓度为200、50 mg/L的烟尘悬浊液在第1、3、5、7、9代对细胞染毒,同代对照组不加烟尘悬浊液同步培养.从第10代起,高、低剂量染毒组常规传代直至软琼脂克隆形成检测实验呈阳性.采用透射电镜观察细胞超微结构,以免疫印迹法及免疫组织化学法观察各组细胞肺表面活性蛋白SP(-B)、SP-C、波形蛋白(Vimentin)、结缔组织生长因子(CTGF)、水通道蛋白5(AQP5)表达.结果从形态改变、软琼脂克隆形成实验及肺泡表面活性蛋白(SP)-B、SP-C的表达上发现经烟尘诱导,RLE-6TN细胞超微结构会发生改变,且高、低剂量染毒组克隆形成率与染毒培养代数相关性有统计学意义(P<0.01),但与烟尘染毒水平相关性无统计学意义(P>0.05).免疫组织化学显示高、低剂量染毒组第30代细胞CTGF阳性、SP-B阴性,空白对照组CTGF阴性、SP-B阳性.Western blot示高、低剂量染毒组第25、30、35代细胞及空白对照组Vimentin和AQP5均为阴性.结论云锡炼厂烟尘导致ATⅡ恶性转化模型中CTGF表达阳性,但Vimintin及AQP5的表达呈阴性.Objective The aim of this manuscript is to observe the expression of Vimentin,CTGF and AQP5 in the malignant transformation model which established in vitro to mimic the chronic exposure of alveolar epithelial type Ⅱ cells to Yunnan Tin mine smelting dust.Methods Immortalized alveolar epithelial type Ⅱ cells（RLE- 6TN） were set into three groups. One group was exposed to high concentration of Yunnan Tin mine smeltingdust（200 mg/L） and another group was exposed to low concentration of Yunnan Tin mine smelting dust（50 mg/L）. Both the two groups were exposed to high and low concentration during 1,3,5,7,9 cycles of 72 h. The control group was not treated by the dust. From the 10 th cycle, all cells were transmitted routinely until anchorage independent growth were determined. The ultrastructure of RLE- 6TN cells were observed by electron microscope.Immunocytochemistry（ICC） and western blot（WB） were used to test the expression of SP- B, SP- C,Vimentin、 CTGF, AQP5.Results According to the changes of cell morphology, the result of anchorage independent growth test and the expression of SP- B,SP- C,we find that the ultrastructure of RLE- 6TN could be injured by the dust. At the same time, statistics analysis shows a significant difference could be found between cloning efficiency and cell cycles（P〈0.01）,while there is no statistic relative between cloning efficiency and dust concentration（P〈0.05）. Immunocytochemistry（ICC） and western blot（WB） results were SP- C（-） SP- B（-） CTGF（＋） Vimentin（-） AQP5（-） in both two concentration groups while SP- C（＋） SP- B（＋） CTGF（-） Vimentin（-） AQP5（-） in control group.Conclusions Chronic exposure of alveolar epithelial typeⅡ cells to Yunnan Tin mine smelting dust could lead to RLE- 6 TN cells malignant transformation in vitro and induce the expression of CTGF

关 键 词：肺泡Ⅱ型上皮细胞 RLE-6TN 云锡烟尘 恶性转化 肺表面活性蛋白 结缔组织生长因子 波形蛋白 水通道蛋白5 

分 类 号：R734.2[医药卫生—肿瘤]