个旧矿粉诱导的肺癌细胞中分化抗原9的表达及基因突变分析  

Expression and gene mutation of cluster of differentiation 9 in lung cancer cells induced by mineral powder in Gejiu

作　　者：梁锐[1] 金克炜[1] 王志强[2] 畅继武[3] 马富玲[3] 王一[3] 高倩[1] 

机构地区：[1]昆明医学院病理学教研室,650031 [2]昆明医学院第一附属医院干疗外科,650031 [3]天津市泌尿外科研究所

出　　处：《中华劳动卫生职业病杂志》2007年第9期537-540,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金(30560053);高等学校博士学科点专项科研基金(20050678004)

摘　　要：目的探讨分化抗原9(CD9)在个旧矿粉诱导永生化人支气管上皮细胞(BEAS-2B)恶性转化为肺癌细胞过程中的异常表达和基因突变。方法以BEAS-2B为对照组(20瓶),以个旧矿粉体外诱导BEAS-2B所产生的恶性转化细胞为实验组(20瓶)。采用免疫细胞化学技术检测CD9蛋白在BEAS- 2B和恶性转化细胞中的表达。采用逆转录-聚合酶链反应(RT-PCR)技术检测CD9 mRNA在BEAS-2B和恶性转化细胞中的表达;并对其RT-PCR产物进行DNA序列测定。结果CD9蛋自在BEAS-2B中的表达率(100%,20/20)与其在恶性转化细胞中的表达率(35%,7/20)差异有统计学意义(P<0.01)。CD9 mRNA在BEAS-2B中的表达强度(0.91±0.09)明显高于其在恶性转化细胞中的表达强度(0.34±0.14),差异有统计学意义(P<0.01)。DNA序列测定显示,在BEAS-2B的恶性转化细胞中CD9基因存在2处点突变,在第231位置发生G-→T转换;在第119位置发生T→A转换,而且这2处的替代引起的错译突变均导致了编码氨基酸的改变,第40位氨基酸由谷胺酰胺(Gln,Q)变为组氨酸(His,H);第3位氨基酸由缬氨酸(Val,V)变为天冬氨酸(Asp,D)。结论CD9的表达缺失或下调以及基因突变在个旧矿粉诱导BEAS-2B恶性转化为肺癌细胞的过程中可能发挥重要作用。Objective To investigate the expression and gene mutation of cluster of differentiation 9 （CD9） in the pathway of the mineral powder induced malignant transformation in immortalized human bronchial epithelial cells （BEAS-2B） in Gejiu. Methods BEAS-2B cells served as the control group and its malignant transformation cells induced by mineral powder in Gejiu were considered as experiment group. The expression of CD9 protein in 20 bottles of BEAS-2B cells and 20 bottles of malignant transformation cells was evaluated by immunocytochemistry. The mRNA expression of CD9 in 10 bottles of BEAS-2B cells and 10 bottles of malignant transformation cells was examined by reverse transcriptase polymerase chain reaction （RT-PCR）. Gene mutation was detected in the products of RT-PCR by DNA sequencing. Results There was significant difference between the expression of CD9 protein in BEAS-2B cells （100%, 20/20） and that in its malignant trans- formation cells （35%, 7/20） （P〈0.01）. The expression of CD9 mRNA in BEAS-2B cells （0.91＋0.09） was significantly higher than that in its malignant transformation cells （0.34±0.14） （P〈0.01）. Two point mutation of CD9 gene was detected in the malignant transformation cells of BEAS-2B by DNA sequencing. The change of G→T in the base of 231 led to the change of Gln→His in the amino acids of 40. The change of T→A in the base of 119 led to the change of Val→Asp in the amino acids of 3. Conclusion The absence or down-regnlation of CD9 expression and point mutation in the malignant transformation cells of BEAS-2B may play a considerable role in the pathway of the malignant transformation in the BEAS-2B cells induced by mineral powder in Gejiu.

关 键 词：粉尘 矿物质 分化抗原9 肺肿瘤 基因表达 

分 类 号：R734.2[医药卫生—肿瘤]