CSC诱导BEAS-2B细胞恶性转化中miRNA表达谱变化  

Identification of differentially miRNAs expression profile induced by CSC in malignant transformation of BEAS-2B cells

作　　者：张亚平 张佳彤[2] 杨萨 李中秋 周洋洋 张巧[1] ZHANG Ya-ping;ZHANG Jia-tong;YANG Sa;LI Zhong-qiu;ZHOU Yang-yang;ZHANG Qiao(College of public health,Zhengzhou University,Zhengzhou Henan 450001,China;The hospital of Zhengzhou University,Zhengzhou Henan 450001,China)

机构地区：[1]郑州大学公共卫生学院,河南郑州450001 [2]郑州大学医院,河南郑州450001

出　　处：《毒理学杂志》2018年第6期436-442,共7页Journal of Toxicology

基　　金：国家自然科学基金(81172717)

摘　　要：目的通过烟草烟雾凝集物(cigarette smoke condensate,CSC)慢性染毒永生化人支气管上皮细胞BEAS-2B体外构建恶性转化细胞模型,miRNA芯片技术筛选模型中差异表达的miRNAs,发现与恶性转化相关的miRNAs,为进一步研究CSC诱导恶性转化的作用机制提供实验基础。方法CCK-8检测细胞存活率,确定CSC染毒终浓度。划痕实验、克隆形成实验、MTT实验及凋亡检测评估细胞恶性转化情况。MiRNAs芯片检测BEAS-2B和CSC30组细胞,明确差异表达谱。表达下调的hsa-miR-2115-3p和hsa-miR-4521进行RT-PCR验证。结果CSC诱导的恶性转化细胞迁移能力、克隆形成能力和增殖活力明显增强,凋亡率明显下降。相对于BEAS-2B组,CSC30组细胞中有78种miRANs差异表达,其中52种表达上调,26种表达下调。RT-PCR验证hsa-miR-2115-3p和hsa-miR-4521表达下调与芯片结果一致。结论CSC可诱导BEAS-2B细胞发生恶性转化,采用miRNA芯片技术筛选到miRNA差异表达谱,其可能与CSC诱导恶行转化的发生发展有关。Objective To establish malignant transformation of human bronchial epithelial(BEAS-2 B) cells by chronically infected with cigarette smoke condensate(CSC). To screen and identify the miRNAs differential expressed profile by the miRNA assay to make further study in the mechanism of CSC-induced malignant transformation. Methods Cell viability was detected via CCK-8 assay to determine final concentration of CSC. Scratch test, colony formation test, MTT assay, and apoptosis assay were applied to evaluate the malignant transformation of cells. MiRNAs assay was applied to detect differential expression profile between BEAS-2 B and CSC30 groups. Down-regulated hsa-miR-2115-3 p and hsa-miR-4521 were verified by RT-PCR. Result Cells migration ability, colony formation ability and proliferation activity were significantly increased. In addition, the cell apoptosis rate was significantly decreased. Seventy-eight miRANs were differentially expressed in CSC30 group. Compared with BEAS-2 B group, of which 52 up-regulated and 26 down-regulated miRNAs were observed. RT-PCR confirmed that the down-regulation of hsa-miR-2115-3 p and hsa-miR-4521 was consistent with the microarray result. Conclusion CSC had the ability to induce the malignant transformation of BEAS-2 B cells. MiRNAs differential expression profile of cell model was obtained, which may be related to the occurrence and development of CSC induced malignant transformation.

关 键 词：CSC BEAS-2B MIRNA芯片 MIRNA 

分 类 号：R114[医药卫生—卫生毒理学]