Breast Hormone Concentrations in Random Fine-Needle Aspirates of Healthy Women Associate with Cytological Atypia and Gene Methylation

Abstract

Sex steroid hormones contribute to breast cancer development, but data on concentrations of these within breast tissue are limited. We performed simultaneous multiparameter measurement of breast sex steroids, breast epithelial cytology, and DNA methylation in 119 healthy women (54 pre- and 65 postmenopausal) without a history of breast cancer. Random fine-needle aspiration (rFNA) of the breast was performed simultaneously with blood collection. Breast samples were analyzed by LC/MS-MS for estrone, estradiol, progesterone, androstenedione, and testosterone. Blood samples were assayed for estradiol and progesterone by immunoassay. Cytomorphology was classified using the Masood Score, and DNA methylation of eight genes was analyzed using quantitative multiplexed methylation-specific PCR, and expressed as the cumulative methylation index (CMI). Serum and breast concentrations of estradiol and progesterone showed significant correlation (Spearman = 0.34, = 0.001 and = 0.69, < 0.0006, respectively). Progesterone concentration was significantly higher in the premenopausal breast ( < 0.0008), and showed a luteal surge. Breast estrone and estradiol concentrations did not differ significantly by menopause, but androstenedione concentration was higher in the breasts of postmenopausal women ( = 0.026 and = 0.208). Breast androgens were significantly correlated with breast density (Spearman = 0.27, = 0.02 for testosterone) and CMI (Spearman = 0.3, = 0.038 for androstenedione). Our data indicate that future larger studies of breast steroid hormones along with other parameters are feasible. Significant associations of breast androgen concentrations with breast density and gene methylation warrant future study..

Association of Serum Concentrations of Testosterone and Insulin with the Degree of Cervical Squamous Intraepithelial Lesions in Cuban Women

“…The fact that testosterone is one of the precursors of estrogen synthesis is a doubtful factor in determining the role of androgen concentration in breast carcinogenesis and other cancer sites in women, such as the cervix. It is necessary to measure the levels of steroid hormones at the tissue level to establish the differences between serum and local hormone concentrations [55,56], and thus obtain valuable data on susceptibility to them.…”

The Other Side of the Coin: May Androgens Have a Role in Breast Cancer Risk?

“…However, since most of the studies evaluated circulating T levels that are metabolized in estrogens [ 103 ] and the effects of T on mammary cell proliferation are closely related to 5α-reductase and aromatase tissue expression and activity, these evidences cannot totally reflect the hormone concentration within normal breast tissue because the gland follows its own specific intracrinology [ 32 ]. Therefore, a promising method to measure breast steroid hormone levels could be the fine nipple aspiration that, allowing to distinguish the differences between local and serum hormone concentrations [ 104 , 105 ], might provide helpful data to assess breast cancer susceptibility. Moreover, AR expression was not assessed in these studies, making it difficult to understand if AR activation might promote, or not, the malignant transformation.…”

Measurement of Breast Tissue Estrogens by Liquid Chromatography-tandem Mass Spectrometry

“…), после раствор переносили в стеклянные виалы для анализа. Подготовку образцов проводили по методике, описанной ранее [11] Рисунок 1. Абсолютные площади пиков эстрадиола (A) и эстрона (Б) в трех участках одного биоптата, для каждого из которых использовали три варианта измельчения перед этапом экстракции.…”

“…Сравнение методов нормировки на массу образца ткани или массу липидного остатка, которые были ранее описаны в работах [11,12], выполнено с использованием трех образцов примерно одинаковой массы, взятых из трех разных частей биоптата от одного пациента и проанализированы. Нормированные площади пиков эстрона и эстрадиола представлены в таблице 4.…”

“…Предыдущие работы по измерению эстрогенов в образцах ткани молочной железы (МЖ) с помощью ВЭЖХ-МС были сфокусированы на анализе тонкоигольных аспиратов ткани МЖ [11], образцов жировой ткани МЖ [12], и клеточной культуры РМЖ [13]. Тем не менее, разработка метода исследования ткани МЖ с быстрым приготовлением образцов, не требующим использования трудоемких процедур экстракции, остается многообещающей.…”