Tamoxifen induces pleiotrophic changes in mammary stroma resulting in extracellular matrix that suppresses transformed phenotypes.
Tamoxifen induces pleiotrophic changes in mammary stroma resulting in extracellular matrix that suppresses transformed phenotypes.
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DOI：
10.1186/bcr2220
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Schedin P
中科院分区：
文献类型：
--
作者：
Hattar R;Maller O;McDaniel S;Hansen KC;Hedman KJ;Lyons TR;Lucia S;Wilson RS Jr;Schedin P
乳腺的功能单位被定义为上皮细胞及其微环境，这一假设预测上皮细胞功能的变化将伴随着乳腺基质的同时变化。为了验证这一假设，我们研究了抗雌激素药物他莫昔芬是否会改变乳腺间质的功能。将40只70日龄的雌性大鼠随机分为两组，每组20只，每天皮下注射1.0 mg/kg他莫昔芬或溶媒，持续30天，随后为3天洗脱期。收集乳腺组织，测定他莫昔芬对乳腺上皮和间质的影响。正如预期，他莫昔芬抑制乳腺肺泡发育和乳腺上皮细胞增殖。与对照成纤维细胞相比，从他莫昔芬处理的大鼠中分离的原代乳腺成纤维细胞显示出运动性降低三倍，并且在其基质中掺入较少的纤连蛋白;指示成纤维细胞静止的属性。巨噬细胞溶酶体标记物CD 68的免疫组织化学分析表明，他莫昔芬处理的大鼠乳腺中巨噬细胞浸润减少。通过质谱的蛋白质组学分析鉴定了几种细胞外基质（ECM）蛋白，其表达水平与他莫昔芬治疗通过Western印迹验证。从他莫昔芬治疗的大鼠乳腺组织有减少纤维连接蛋白和增加胶原蛋白1水平。此外，ECM蛋白水解在他莫昔芬治疗的大鼠中减少，如通过纤连蛋白、层粘连蛋白1、层粘连蛋白5和胶原1切割片段的减少所检测到的。与他莫昔芬治疗抑制ECM蛋白水解一致，基质金属蛋白酶-2水平和活性降低。从他莫昔芬治疗的大鼠中生物化学提取的乳腺ECM抑制了体外巨噬细胞的运动，这是通过加入蛋白水解的胶原蛋白或纤连蛋白来拯救的。来自他莫昔芬处理的大鼠的乳腺ECM还抑制乳腺肿瘤细胞的运动性、侵袭性和趋合性，在三维培养中减小类器官的大小，并在原位异种移植模型中阻断肿瘤促进;通过添加外源性纤连蛋白可以部分逆转这种效应。这些数据支持这一假设，即乳腺间质对他莫昔芬治疗的反应与上皮细胞一致，并重塑为抑制肿瘤细胞进展的微环境。纤连蛋白水平降低和ECM周转减少似乎是静止乳腺微环境的标志。这些数据可以提供对促进肿瘤休眠的乳腺微环境的属性的洞察。
The functional unit of the mammary gland has been defined as the epithelial cell plus its microenvironment, a hypothesis that predicts changes in epithelial cell function will be accompanied by concurrent changes in mammary stroma. To test this hypothesis, the question was addressed of whether mammary stroma is functionally altered by the anti-oestrogen drug tamoxifen. Forty female rats at 70 days of age were randomised to two groups of 20 and treated with 1.0 mg/kg tamoxifen or vehicle subcutaneously daily for 30 days, followed by a three-day wash out period. Mammary tissue was harvested and effects of tamoxifen on mammary epithelium and stroma determined. As expected, tamoxifen suppressed mammary alveolar development and mammary epithelial cell proliferation. Primary mammary fibroblasts isolated from tamoxifen-treated rats displayed a three-fold decrease in motility and incorporated less fibronectin in their substratum in comparison to control fibroblasts; attributes indicative of fibroblast quiescence. Immunohistochemistry analysis of CD68, a macrophage lysosomal marker, demonstrated a reduction in macrophage infiltration in mammary glands of tamoxifen-treated rats. Proteomic analyses by mass spectrometry identified several extracellular matrix (ECM) proteins with expression levels with tamoxifen treatment that were validated by Western blot. Mammary tissue from tamoxifen-treated rats had decreased fibronectin and increased collagen 1 levels. Further, ECM proteolysis was reduced in tamoxifen-treated rats as detected by reductions in fibronectin, laminin 1, laminin 5 and collagen 1 cleavage fragments. Consistent with suppression in ECM proteolysis with tamoxifen treatment, matrix metalloproteinase-2 levels and activity were decreased. Biochemically extracted mammary ECM from tamoxifen-treated rats suppressed in vitro macrophage motility, which was rescued by the addition of proteolysed collagen or fibronectin. Mammary ECM from tamoxifen-treated rats also suppressed breast tumour cell motility, invasion and haptotaxis, reduced organoid size in 3-dimensional culture and blocked tumour promotion in an orthotopic xenograft model; effects which could be partially reversed by the addition of exogenous fibronectin. These data support the hypothesis that mammary stroma responds to tamoxifen treatment in concert with the epithelium and remodels to a microenvironment inhibitory to tumour cell progression. Reduced fibronectin levels and reduced ECM turnover appear to be hallmarks of the quiescent mammary microenvironment. These data may provide insight into attributes of a mammary microenvironment that facilitate tumour dormancy.
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影响因子：
11.2
作者：
Barkan D;Kleinman H;Simmons JL;Asmussen H;Kamaraju AK;Hoenorhoff MJ;Liu ZY;Costes SV;Cho EH;Lockett S;Khanna C;Chambers AF;Green JE
通讯作者：
Green JE
影响因子：
--
作者：
Buck, MB;Pfizenmaier, K;Knabbe, C
通讯作者：
Knabbe, C
影响因子：
4.4
作者：
Adair-Kirk, TL;Atkinson, JJ;Senior, RM
通讯作者：
Senior, RM
影响因子：
50.3
作者：
Allinen, M;Beroukhim, R;Polyak, K
通讯作者：
Polyak, K
影响因子：
3.8
作者：
JORDAN, VC
通讯作者：
JORDAN, VC