【作者】 宾晓农； 谭敏； 吕嘉春； 纪卫东； 陈家堃； 蒋义国；

【Author】 Bin Xiaonong,Tan Min,Lu Jiachun,Ji Weidong,Chen Jiakung Guangzhou medical college,Guangzhou,510182,China

【机构】 广州医学院化学致癌研究所； 广州中医药大学第二附属医院病理科；

【摘要】 目的:探讨用BPDE和NiS转化的16HBE恶性细胞系BPDE-16HBE和NiS-16HBE的致癌性和转移潜能,并鉴定其生物学差异。方法:应用单个细胞克隆技术分离建立BPDE、NiS转化的16HBE恶性细胞株BPDE-16HBE和NiS-16HBE;基因芯片技术检测BPDE-16HBE和NiS-16HBE细胞株基因差异表达种类和数量;RT-PCR法分析E-cadherin的mRNA的表达和E-cadherin启动子甲基化状况;MTT法、平板法、boyden小室法、细胞染色体分型法以及动物实验检测BPDE-16HBE和NiS-16HBE细胞株的体内外生物学特征。结果:①成功分离建立了具有转移能力的BPDE-16HBE和NiS-16HBE恶性细胞系;②基因芯片技术检测BPDE-16HBE和NiS-16HBE恶性细胞系,呈现差异表达的基因数量有22个,其中11个(50%)下调,另11个(50%)上调;③BPDE-16HBE恶性细胞系E-cadherin的mRNA的表达水平低于NiS-16HBE恶性细胞系,E-cadherin启动子甲基化高于NiS-16HBE恶性细胞系;④BPDE-16HBE恶性细胞系的体外增殖能力、克隆形成率和体外侵袭性较NiS-16HBE恶性细胞系高;⑤BPDE-16HBE恶性细胞系体内成瘤性和转移性较NiS-16HBE高;⑥BPDE-16HBE和NiS-16HBE恶性细胞系染色体众数比较无差异。结论:BPDE-16HBE和NiS-16HBE恶性细胞系具有不同的生物学特性和分子生物学特性;BPDE-16HBE恶性细胞系高致癌性和转移潜能可能与E-cadherin的mRNA的表达水平低,而E-cadherin启动子高甲基化有关。更多还原

【Abstract】 Objective To explore the possibility of separating and establishing clonal transforming cell subpopulations with different metastatic phonetype from human bronchial epithelial cell line(16HBE)treated with BPDE and NiS,to identify the difference of biological and molecular biological between BPDE-16HBE and NiS-16HBE cell line.Methods Two transforming subpopulations cell lines(BPDE-16HBE and NiS-16HBE)were isolated and established by the single cell cloning techniques from 16HBE treated with BPDE and NiS.Gene chip patten,mRNA expression were detected in BPDE-16HBE and NiS-16HBE by cDNA microarray,RT-PCR.Biological characteristic of vivo and vitro were determined in BPDE-16HBE and NiS-16HBE cell lines by MTT, plate,Boyden chamber methods and animal models.Results①Two sub-cell lines,BPDE-16HBE and NiS-16HBE which had different metastatic phonetype,were successfully isolated and established from 16HBE treated with BPDE and NiS.②Twenty-two differential expressed genes were obtained, Among these genes,11 were up-regulated and 11 were down-regulated.③BPDE-16HBE cell line had lower level of mRNA expression of E-cadherin than those of NiS-16HBE cell lines.Aberrant DNA methylation(promoter hypermethylation)of E-cadherin in BPDE-16HBE cell line was identified signficantly higher than in NiS-16HBE cell lines.④The proliferation,clone formation and vitro invastion of BPDE-16HBE cell line was signficantly higher than in NiS-16HBE cell lines.⑤The tumorigenicity and metastasis rate in nude mouse of BPDE-16HBE cell line was remarkably higher than those of NiS-16HBE cell line.⑥No significant difference of the chromosome number was observed between BPDE-16HBE and NiS-16HBE cell lines.Conclusion BPDE-16HBE and NiS-16HBE cell lines established from human bronchial epithelial cell line treated with BPDE and NiS have different biological and molecular characteristics.The high invasion and metastasis ability of BPDE-16HBE cell line might be related to both the lower level ofmRNA expression and the promoter hypermethylation of E-cadherin gene.更多还原