Tamoxifen induces expression of immune response-related genes in cultured normal human mammary epithelial cells.
Tamoxifen induces expression of immune response-related genes in cultured normal human mammary epithelial cells.
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DOI：
10.1158/0008-5472.can-08-2806
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发表时间：
2009-02-01
期刊：
影响因子：
11.2
通讯作者：
Poirier MC
中科院分区：
文献类型：
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作者：
Schild-Hay LJ;Leil TA;Divi RL;Olivero OA;Weston A;Poirier MC
使用他莫昔芬（TAM）与乳腺癌发病率降低50%和子宫内膜癌发病率增加有关。在这里，我们记录了tam诱导的正常人乳腺上皮细胞（NHMEC菌株编号5、16和40）的基因表达变化，这些细胞是从3个个体的乳房缩小成形术中提取的组织中建立的。细胞暴露于0、10或50 μM的TAM中48小时，采用TAM-DNA（经dG-N2-TAM修饰的DNA）化学发光免疫分析法（CIA）检测(E)-α-（脱氧鸟苷- n2 -酰基）-他莫西芬（dG-N2-TAM）加合物的形成，采用NCI DNA寡核苷酸芯片和实时(RT)- pcr检测基因表达变化。48小时时，10 μM和50 μM TAM的细胞存活率分别为85.6%和48.4%，未检测到dG-N2-TAM加合物。对于微阵列，细胞暴露在10 μM TAM中，表达变化≥3倍的基因如下：菌株16有13个基因上调，1个基因下调；菌株5有17个基因上调；菌株40有11个基因表达上调。干扰素诱导基因（IFITM1、IFIT1、IFNA1、MXI和GIP3）和钾离子通道（KCNJ1）均上调。雌激素和外源代谢相关基因的表达未见明显变化。RT-PCR结果显示干扰素α (IFNA1)上调，证实了tam诱导的微阵列鉴定基因上调，但GIP3和MX1在菌株40中未上调。在三种NHMEC菌株中诱导干扰素相关基因表明，除了激素作用外，TAM暴露还可能增强正常乳腺组织的免疫反应。
Use of tamoxifen (TAM) is associated with a 50% reduction in breast cancer incidence and an increase in endometrial cancer incidence. Here, we documented TAM-induced gene expression changes in cultured normal human mammary epithelial cells (NHMEC strains numbered 5, 16 and 40), established from tissue taken at reduction mammoplasty from 3 individuals. Cells exposed to 0, 10 or 50 μM TAM for 48 hours were evaluated for (E)-α-(deoxyguanosin-N2-yl)-tamoxifen (dG-N2-TAM) adduct formation by TAM-DNA (DNA modified with dG-N2-TAM) chemiluminescence immunoassay (CIA), gene expression changes using NCI DNA-oligonucleotide microarray, and real time (RT)-PCR. At 48 hr, cells exposed to 10 μM and 50 μM TAM were 85.6% and 48.4% viable, respectively, and there were no measurable dG-N2-TAM adducts. For microarray, cells were exposed to 10 μM TAM and genes with expression changes of ≥ 3-fold were as follows: thirteen genes up-regulated and one down-related for strain 16; seventeen genes up-regulated for strain 5; and eleven genes up-regulated for strain 40. Interferon-inducible genes (IFITM1, IFIT1, IFNA1, MXI and GIP3), and a potassium ion channel (KCNJ1) were up-regulated in all 3 strains. No significant expression changes were found for genes related to estrogen or xenobiotic metabolism. RT-PCR revealed up-regulation of interferon α (IFNA1) and confirmed the TAM-induced up-regulation of the genes identified by microarray, with the exception of GIP3 and MX1, which were not up-regulated in strain 40. Induction of interferon-related genes in the three NHMEC strains suggests that, in addition to hormonal effects, TAM exposure may enhance immune response in normal breast tissue.