Banerjee Nilasha, Miller Naomi, Allen Christine, Bendayan Reina
Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, 144 College Street, Toronto, ON, M5S 3M2, Canada.
Breast Cancer Res Treat. 2014 Jun;145(3):647-61. doi: 10.1007/s10549-014-2990-y. Epub 2014 May 16.
Two-thirds of newly diagnosed hormone-dependent (HR?) breast cancers are detected in post-menopausal patients where estrone-3-sulphate (E3S) is the predominant source for tumour estradiol. Understanding intra-tumoral fate of E3S would facilitate in the identification of novel molecular targets for HR? post-menopausal breast cancer patients. Hence this study investigates the clinical expression of (i) organic anion-transporting polypeptides (OATPs), (ii) multidrug resistance protein (MRP-1), breast cancer resistance proteins (BCRP), and (iii) sulphatase (STS), 17β-hydroxysteroid dehydrogenase (17β-HSD-1), involved in E3S uptake, efflux and metabolism, respectively. Fluorescent and brightfield images of stained tumour sections (n = 40) were acquired at 4× and 20× magnification, respectively. Marker densities were measured as the total area of positive signal divided by the surface area of the tumour section analysed and was reported as % area (ImageJ software). Tumour, stroma and non-tumour tissue areas were also quantified (Inform software), and the ratio of optical intensity per histologic area was reported as % area/tumour, % area/stroma and % area/non-tumour. Functional role of OATPs and STS was further investigated in HR? (MCF-7, T47-D, ZR-75) and HR-(MDA-MB-231) cells by transport studies conducted in the presence or absence of specific inhibitors. Amongst all the transporters and enzymes, OATPs and STS have significantly (p < 0.0001) higher expression in HR? tumour sections with highest target signals obtained from the tumour regions of the tissues. Specific OATP-mediated E3S uptake and STS-mediated metabolism were also observed in all HR? breast cancer cells. These observations suggest the potential of OATPs as novel molecular targets for HR? breast cancers.
三分之二新诊断的激素依赖性(HR?)乳腺癌是在绝经后患者中检测到的,其中硫酸雌酮(E3S)是肿瘤雌二醇的主要来源。了解E3S在肿瘤内的命运将有助于识别HR?绝经后乳腺癌患者的新型分子靶点。因此,本研究调查了(i)有机阴离子转运多肽(OATP)、(ii)多药耐药蛋白(MRP-1)、乳腺癌耐药蛋白(BCRP)以及(iii)硫酸酯酶(STS)、17β-羟基类固醇脱氢酶(17β-HSD-1)的临床表达,它们分别参与E3S的摄取、外排和代谢。分别在4倍和20倍放大倍数下获取染色肿瘤切片(n = 40)的荧光和明场图像。标记物密度通过阳性信号总面积除以分析的肿瘤切片表面积来测量,并报告为面积百分比(ImageJ软件)。还对肿瘤、基质和非肿瘤组织区域进行了定量(Inform软件),每个组织学区域的光强度比报告为%面积/肿瘤、%面积/基质和%面积/非肿瘤。通过在存在或不存在特异性抑制剂的情况下进行转运研究,进一步研究了OATP和STS在HR?(MCF-7、T47-D、ZR-75)和HR-(MDA-MB-231)细胞中的功能作用。在所有转运蛋白和酶中,OATP和STS在HR?肿瘤切片中的表达显著更高(p < 0.0001),从组织的肿瘤区域获得最高的靶信号。在所有HR?乳腺癌细胞中也观察到了特异性OATP介导的E3S摄取和STS介导的代谢。这些观察结果表明OATP作为HR?乳腺癌新型分子靶点的潜力。