环境与职业医学2018，Vol.35Issue(4)：316-322,7.DOI:10.13213/j.cnki.jeom.2018.17584
苯并[α]芘对人支气管上皮细胞H19与SAHH表达的影响
Effects of benzo[α]pyrene on expression of H19 and SAHH in human bronchial epithelial cells
摘要
Abstract
[Objective] To investigate the effects of benzo[a]pyrene (BaP) on the expression of long non-coding RNA (IncRNA) H19 and S-adenosylhomocysteine hydrolase (SAHH) in human bronchial epithelial cells (16HBE),and to assess the relationship between H19 and SAHH.[Methods] A batch of 16HBE cells were exposed to BaP at 1,2,4,8,16,and 32μmol/L for 24h,and a control group (culture medium only) and a solvent control group were established.Another batch of 16HBE cells were exposed to 16 μnol/L BaP for 1,2,4,8,12,and 24h,and a control group (0h) was also established.The relative expression of H19 was detected by reverse transeriptionPCR (RT-PCR).The protein expression of SAHH was detected by Western blot.The distribution of H19 and SAHH were detected by lncRNA fluorescence in situ hybridization (FISH) combined with laser scanning confocal microscopy.[Results] The expression levels of H19 were not different between the solvent control group and the untreated group.The relative expression levels of H19 in the 16HBE cells were gradually elevated with higher BaP concentrations and extended exposure time,according to the results of RT-PCR.The expression levels of H19 were significantly higher in the 16HBE ceils exposed to BaP at concentrations of 8,16,and 32 μmol/L for 24 h than those of the solvent control group (Ps ＜ 0.05),and higher in the 16HBE cells exposed to 16 μmol/L BaP for 4,8,12,and 24 h than those of the 0 h group (Ps ＜ 0.05).The relative expression levels of H19 reached a peak at 32 μmol/L BaP,which increased 78.0％ compared with the solvent control group.For 16 μmol/L BaP exposure,the relative expression levels of H19 reached a peak at 24h,which increased 48.3％ compared with the 0h group.The protein expression levels of SAHH in 16HBE were not different between the solvent control group and the untreated group,and significantly decreased with higher BaP concentrations and extended exposure time (P ＜ 0.05),according to the results of Western blot.The protein expression levels of SAHH reached a bottom at 32 μmol/L BaP,which was 37.2％ lower than that of the solvent control group.For 16 μmol/L BaP exposure,the protein expression levels of SAHH reached a bottom at 24h,which was 33.1％ lower than that of the 0h group.The immunofluorescence results showed that both H19 and SAHH were distributed in cytoplasm and nucleus,predominantly in perinuclear cytoplasm and nuclear membrane.In the 16HBE cells exposed to 16 μmol/L BaP for 24 h,more H 19 were detected in perinuclear cytoplasm,while less SAHH were detected as compared to the 0h group,and the immunofluorescence intensity of H 19 and SAHH co-localized in cytoplasm and nucleus was increased.[Conclusion] Exposure to BaP could increase the relative expression levels of H19 in 16HBE cells,while decrease the protein expression levels of SAHH.Co-localization of H19 and SAHH in 16HBE cells suggests an interaction between H19 and SAHH.关键词
苯并[α]芘/长链非编码RNA H19/逆转录-聚合酶链式反应/S-腺苷高半胱氨酸水解酶/原位荧光杂交Key words
Benzo[a]pyrene/long non-coding RNA H19/reverse transcription-PCR/S-adenosylhomocysteine hydrolase/fluorescence in situ hybridization分类
医药卫生引用本文复制引用
傅晔,杨瑾,王武斌,张红杰,刘艳丽,牛莹莹..苯并[α]芘对人支气管上皮细胞H19与SAHH表达的影响[J].环境与职业医学,2018,35(4):316-322,7.基金项目
山西省自然科学基金项目(编号:201701D121146) （编号:201701D121146）
国家自然科学基金项目(编号:81273041) （编号:81273041）
