Epidermal growth factor receptor signalling in human breast cancer cells operates parallel to estrogen receptor α signalling and results in tamoxifen insensitive proliferation.
Epidermal growth factor receptor signalling in human breast cancer cells operates parallel to estrogen receptor α signalling and results in tamoxifen insensitive proliferation.
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人乳腺癌细胞中的表皮生长因子受体信号传导平行于雌激素受体α信号传导，并导致他莫昔芬不敏感增殖。
DOI：
10.1186/1471-2407-14-283
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发表时间：
2014-04-23
期刊：
影响因子：
3.8
通讯作者：
Meerman JH
中科院分区：
文献类型：
--
作者：
Moerkens M;Zhang Y;Wester L;van de Water B;Meerman JH
三苯氧胺耐药是雌激素受体（ER）α阳性乳腺癌患者治疗中的主要问题。虽然他莫昔芬耐药背后的机制尚未完全了解，临床数据表明，受体酪氨酸激酶的表达增加参与。在这里，我们研究了人乳腺癌MCF 7细胞的雌激素和抗雌激素敏感性，该细胞具有适度的、逆转录病毒介导的、异位表达的表皮生长因子受体（MCF 7-EGFR）。MCF 7-EGFR和亲本细胞的增殖由17β-雌二醇（E2）、表皮生长因子（EGF）或它们的组合诱导。在这些条件下，使用10-12至10-6 M的4-羟基-他莫昔芬（TAM）或氟维司群研究增殖抑制。在不同时间点裂解细胞，检测EGFR、MAPK 1/3、AKT的磷酸化状态及ERα的表达。使用smartpool siRNA建立靶基因的敲低。在用生长因子刺激后6小时，使用Affytron HG-U133 PM阵列板进行转录组学分析。MCF 7-EGFR细胞的增殖可被EGF或E2所诱导。TAM或氟维司群处理显著抑制单独E2刺激的MCF 7-EGFR细胞的增殖。EGF处理E2/TAM处理的细胞导致显著的细胞增殖，从而推翻抗雌激素介导的细胞增殖抑制。然而，在这些条件下，TAM仍然抑制ERα介导的转录。在TAM/E2/EGF条件下，siRNA介导的EGFR敲低抑制EGF驱动的增殖，而ERα敲低则没有。由条件性EGFR信号传导介导的TAM抗性细胞增殖可能依赖于PI 3 K/Akt通路而不是MEK/MAPK通路，因为MEK抑制剂（U 0126）不阻断增殖。在各种E2/TAM/EGF条件下的转录组学分析显示，E2和EGF依赖性转录几乎没有重叠，而是以平行方式操作。我们的数据表明增强的EGFR驱动的信号传导足以推翻TAM介导的对E2驱动的细胞增殖的抑制。这可能对EGFR水平升高的ER阳性乳腺癌的抗雌激素治疗具有深远的意义。
Tamoxifen resistance is a major problem in the treatment of estrogen receptor (ER) α -positive breast cancer patients. Although the mechanisms behind tamoxifen resistance are still not completely understood, clinical data suggests that increased expression of receptor tyrosine kinases is involved. Here, we studied the estrogen and anti-estrogen sensitivity of human breast cancer MCF7 cells that have a moderate, retroviral-mediated, ectopic expression of epidermal growth factor receptor (MCF7-EGFR). Proliferation of MCF7-EGFR and parental cells was induced by 17β-estradiol (E2), epidermal growth factor (EGF) or a combination of these. Inhibition of proliferation under these conditions was investigated with 4-hydroxy-tamoxifen (TAM) or fulvestrant at 10-12 to 10-6 M. Cells were lysed at different time points to determine the phosphorylation status of EGFR, MAPK1/3, AKT and the expression of ERα. Knockdown of target genes was established using smartpool siRNAs. Transcriptomics analysis was done 6 hr after stimulation with growth factors using Affymetrix HG-U133 PM array plates. While proliferation of parental MCF7 cells could only be induced by E2, proliferation of MCF7-EGFR cells could be induced by either E2 or EGF. Treatment with TAM or fulvestrant did significantly inhibit proliferation of MCF7-EGFR cells stimulated with E2 alone. EGF treatment of E2/TAM treated cells led to a marked cell proliferation thereby overruling the anti-estrogen-mediated inhibition of cell proliferation. Under these conditions, TAM however did still inhibit ERα- mediated transcription. While siRNA-mediated knock-down of EGFR inhibited the EGF- driven proliferation under TAM/E2/EGF condition, knock down of ERα did not. The TAM resistant cell proliferation mediated by the conditional EGFR-signaling may be dependent on the PI3K/Akt pathway but not the MEK/MAPK pathway, since a MEK inhibitor (U0126), did not block the proliferation. Transcriptomic analysis under the various E2/TAM/EGF conditions revealed that E2 and EGF dependent transcription have little overlap and rather operate in a parallel fashion. Our data indicate that enhanced EGFR-driven signalling is sufficient to overrule the TAM- mediated inhibition of E2-driven cell proliferation. This may have profound implications for the anti-estrogen treatment of ER-positive breast cancers that have increased levels of EGFR.
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影响因子：
5.3
作者：
Ishida, S;Huang, E;Nevins, JR
通讯作者：
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