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[目的]探讨苯并[α]芘(BaP)对人支气管上皮细胞(16HBE)长链非编码RNA(IncRNA)H19和S-腺苷高半胱氨酸水解酶(SAHH)表达的影响,并初步探究H19和SAHH的关系.[方法]用不同浓度(1、2、4、8、16、32.μmol/L)BaP染毒16HBE细胞24h,设置未处理组(仅加培养液)和溶剂对照组;以16 μmol/L BaP染毒16HBE细胞不同时间(1、2、4、8、12、24h),以0h为对照组.用实时荧光定量逆转录-聚合酶链式反应(RT-PCR)检测H19的表达水平;通过免疫印迹试验(Western blot)检测SAHH的蛋白表达水平;采用lncRNA原位荧光杂交(FISH)技术结合激光共聚焦显微镜观察H19和SAHH的定位.[结果] RT-PCR结果显示,溶剂对照组与未处理组相比,H19表达差异无统计学意义.随着染毒浓度和时间的增加,16HBE细胞H19表达呈现逐渐升高的趋势.8、16、32μmol/L BaP染毒24h及16 μmol/L BaP染毒4、8、12、24h时,H19表达量分别高于溶剂对照组和0h组(均P＜0.05).BaP染毒浓度为32 μtmol/L时,H19表达量达到峰值,较溶剂对照组增加了78.0％;16μmol/L BaP染毒24h时,H19表达量达到峰值,较0h组增加了48.3％.Western blot结果显示,溶剂对照组与未处理组的SAHH表达差异无统计学意义.随着染毒时间和浓度的增加,16HBE细胞SAHH表达逐渐降低(P＜0.05).BaP浓度为32μmol/L时,SAHH表达降至最低,较溶剂对照组降低了37.2％;16μmol/L BaP染毒24h时,SAHH表达降至最低,较0h组降低了33.1％.免疫荧光结果显示,H19和SAHH均在细胞质及细胞核中表达,表达主要分布在核周质及核膜;与0h组相比,16 μmol/L BaP染毒细胞24h后H19在核周质处的荧光加强,而SAHH的荧光明显减弱,两者在胞质、胞核中共定位的荧光强度增加.[结论] BaP染毒可引起16HBE细胞H19表达升高而SAHH表达降低;H19与SAHH在细胞内的共定位表达提示两者可能存在相互作用.“,”[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.