Horne Andrew W, Brown Jeremy K, Nio-Kobayashi Junko, Abidin Hazirah B Z, Adin Zety E H A, Boswell Lyndsey, Burgess Stewart, Lee Kai-Fai, Duncan W Colin
MRC Centre for Reproductive Health, The Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, United Kingdom.
MRC Centre for Reproductive Health, The Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, United Kingdom ; Laboratory of Histology and Cytology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
PLoS One. 2014 Feb 20;9(2):e89400. doi: 10.1371/journal.pone.0089400. eCollection 2014.
Epidemiological studies have shown that cigarette smoking is a major risk factor for tubal ectopic pregnancy but the reason for this remains unclear. Here, we set out to determine the effect of smoking on Fallopian tube gene expression. An oviductal epithelial cell line (OE-E6/E7) and explants of human Fallopian tubes from non-pregnant women (n = 6) were exposed to physiologically relevant concentrations of cotinine, the principle metabolite of nicotine, and changes in gene expression analyzed using the Illumina Human HT-12 array. Cotinine sensitive genes identified through this process were then localized and quantified in Fallopian tube biopsies from non-pregnant smokers (n = 10) and non-smokers (n = 11) using immunohistochemistry and TaqMan RT-PCR. The principle cotinine induced change in gene expression detected by the array analysis in both explants and the cell line was significant down regulation (P<0.05) of the pro-apoptotic gene BAD. We therefore assessed the effect of smoking on cell turnover in retrospectively collected human samples. Consistent with the array data, smoking was associated with decreased levels of BAD transcript (P<0.01) and increased levels of BCL2 transcript (P<0.05) in Fallopian tube biopsies. BAD and BCL2 specific immunolabelling was localized to Fallopian tube epithelium. Although no other significant differences in levels of apoptosis or cell cycle associated proteins were observed, smoking was associated with significant changes in the morphology of the Fallopian tube epithelium (P<0.05). These results suggest that smoking may alter tubal epithelial cell turnover and is associated with structural, as well as functional, changes that may contribute to the development of ectopic pregnancy.
流行病学研究表明,吸烟是输卵管异位妊娠的主要风险因素,但其原因尚不清楚。在此,我们着手确定吸烟对输卵管基因表达的影响。将输卵管上皮细胞系(OE-E6/E7)和来自非孕妇女性(n = 6)的人输卵管外植体暴露于生理相关浓度的可替宁(尼古丁的主要代谢产物),并使用Illumina Human HT-12芯片分析基因表达的变化。然后,通过免疫组织化学和TaqMan RT-PCR对通过该过程鉴定出的可替宁敏感基因在非妊娠吸烟者(n = 10)和非吸烟者(n = 11)的输卵管活检组织中进行定位和定量。芯片分析在体外培养物和细胞系中检测到的可替宁诱导的主要基因表达变化是促凋亡基因BAD的显著下调(P<0.05)。因此,我们评估了吸烟对回顾性收集的人体样本中细胞更新的影响。与芯片数据一致,吸烟与输卵管活检组织中BAD转录本水平降低(P<0.01)和BCL2转录本水平升高(P<0.05)相关。BAD和BCL2特异性免疫标记定位于输卵管上皮。虽然未观察到凋亡或细胞周期相关蛋白水平的其他显著差异,但吸烟与输卵管上皮形态的显著变化相关(P<0.05)。这些结果表明,吸烟可能会改变输卵管上皮细胞更新,并与可能导致异位妊娠发生的结构和功能变化相关。