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PubMed1996

Angiotensin-converting enzyme and enkephalinase in human breast cyst fluid.

Palpable breast cysts with an apocrine epithelial lining (type 1) are reported to be associated with a higher risk of developing breast cancer. The composition of breast cyst fluid (BCF) might include those factors involved in this increased risk. In this study peptidase activities that were active against the substrate [125I]metenkephalin-Arg-Phe were detected in BCF. The products were identified by reversed phase high-performance liquid chromatography (HPLC) as [125I]Tyr-Gly-Gly and [125I]Met-enkephalin. This proteolysis was not inhibited by PCMB, pepstatin A, leupeptin or aprotinin but was by EDTA, showing that the activity was due to metalloproteases. The production of [125I]Try-Gly-Gly was inhibited by phosphoramidon and thiorphan, whereas that of [125I]met-enkephalin was inhibited by captopril and Bothrops jararaca peptide, indicating that these activities are enkephalinase and angiotensin-converting enzyme (ACE) respectively. A fluorometric assay for ACE demonstrated that ACE levels are significantly higher in type 2 BCF than in type 1 BCF (30.8 vs 6.1 nmol hr-1 10 microliters-1, P < 0.001). As the increased risk of cancer is linked to type 1 cysts it is possible that higher levels of peptidase in type 2 BCF reflect a protective environment in the breast in which mitogenic peptide growth factors are neutralised by proteolysis.

Amino Acid Sequence↗
PubMed2002

[Relationship between estrogen, progesterone, total testosterone, androstenedione, and epidural growth factor in blood and breast cyst fluid].

In breast cyst fluid obtained from 50 patients age (47.1 +/- 2.3) sodium, potassium, oestrone (E1), oestradiol (E2) androgens, progesterone, (P) and Epidermal Growth Factor--EGF were determined. Markedly higher oestrogens, progesterone, testosterone, and EGF-levels were detected in cyst fluid than in blood. These data have confirmed previous results and have clearly shown that breast cyst fluid with high potassium (ratio K/Na > 1.0) contains more oestrogens, progesterone, testosterone, androstenedione and EGF than the group with low electrolyte ratio. This finding might be put in correlation with the fact that patients with high ratio K/Na may a higher risk of breast cancer than those with low electrolyte ratio.

Adult↗
PubMed1999

Prostate-specific antigen found in type I breast cyst fluids is a secretory product of the apocrine cells lining breast gross cysts.

Prostate-specific antigen (PSA), a serine protease thought to be exclusively produced by the prostate epithelial cells, has been recently found in human breast tissues and fluids. PSA in breast cancer is associated with the presence of steroid-hormones and receptors, and its presence seems to be a favourable prognostic indicator. In order to clarify whether the cells lining breast cysts may represent the source of PSA found in human breast cyst fluid, we performed an ultrastructural immunolocalization of PSA in the cells surrounding Type I breast cysts, obtained from breast cyst fluids of women affected by breast gross cystic disease, the most commonly occurring benign breast lesions associated with increased cancer risk. These apocrine cells show morphological features typical of actively synthesizing and secreting cells, and a PSA labelling distributed on free ribosomes, RER cisternae, and secretory granules, indicating that the metabolically active apocrine cells lining the Type I cysts are responsible for the production and secretion of PSA in Type I breast cyst fluids. The synthesis and intracystic accumulation of this serine protease in biosynthetically active apocrine Type I cysts can play an important role in the natural history of breast gross cystic disease as well as in the mechanism of cyst evolution.

Adult↗
PubMed1994

Oncostatin M, interleukin 2, interleukin 6 and interleukin 8 in breast cyst fluid.

Women with breast cysts lined by apocrine epithelium (intracystic Na/K < 3) may have a higher risk of developing breast cancer than those whose breast cysts are lined by "flattened" epithelium (intracystic Na/K < 3). In this study, the concentrations of the cytokines, oncostatin M (OSM), interleukin 2 (IL-2), interleukin 6 (IL-6) and interleukin 8 (IL-8) were measured in breast cyst fluid, OSM, IL-2 and IL-6 having been shown to have growth-inhibitory actions on tumour cells. All cytokines were measured by "sandwich" enzyme immunoassays. IL-2 was not detectable in breast cyst fluid. OSM, IL-6 and IL-8 concentrations were significantly higher in the high-electrolyte-ratio group. Although significant positive correlations were found between OSM and IL-6, OSM and IL-8, OSM and Na/K, IL-6 and IL-8, IL-6 and Na/K and IL-8 and Na/K when all samples were analysed together, analysis of the correlations between these analytes in each subgroup separately suggests that the control of production of these cytokines or the mechanism of entry into cyst fluid is likely to be different for the 2 cyst types. The significantly higher intracystic concentrations of OSM and IL-6 in the high-electrolyte-ratio group may partly explain the lower risk of breast cancer in this group of women.

Breast Neoplasms↗
PubMed1999

Prostate specific antigen molecular forms in breast cyst fluid and serum of women with fibrocystic breast disease.

We have analyzed matched serum and breast cyst fluid samples for total PSA from 148 patients with fibrocystic breast disease. We have also determined the molecular forms of PSA (free PSA and PSA bound to alpha1-antichymotrypsin) in 78 breast cyst fluid samples. We found that total PSA can be detected in all cyst fluids and in about 75% of female sera. The median total PSA concentration in breast cyst fluid (bcf) is about 30 times higher than the median in the corresponding sera. Breast cyst fluid and serum PSA are not correlated with each other. Total serum PSA is inversely associated with patient age but the inverse association between bcf PSA and age is weak. Lower total PSA in bcf was seen in women who breast feed, and higher bcf PSA is associated with multiple cysts. Type I cysts (with a high K+/ Na+ ratio) tend to have higher total PSA than Type II cysts. All but three of the fractionated cyst fluids (75/78; 96%) had free PSA as the predominant molecular form. The most consistent finding of our study was the positive association between the cyst fluid K+/Na+ ratio and the free to bound PSA ratio. This association was confirmed by Spearman correlation as well as by Wilcoxon and chi-square analysis. Secretory/apocrine cysts (Type I) tend to have more total PSA and proportionally more free PSA than transudative/flattened cysts (Type II).

Adult↗
PubMed2006

Melatonin and estrogen in breast cyst fluids.

Increased breast cancer risks have been reported among women with gross cystic breast disease (GCBD), although the mechanism for this increase remains unexplained. Relationships between GCBD characteristics, breast cancer risk factors, and the biochemical composition and growth properties of 142 breast cyst fluid (BCF) samples were studied among 93 women with GCBD. Concentrations of melatonin, estrogen (17-beta-estradiol), dehydroepiandrosterone-sulfate (DHEA-S), epidermal growth factor (EGF), transforming growth factor beta (TGF-B1 and TGF-B2), sodium (Na), and potassium (K) were quantified in BCF samples, and human breast cancer cells (MCF-7) were treated with BCF in vitro. Patients were grouped according to BCF Na:K ratios previously linked with increased breast cancer risks (Na:K </= 3, Type 1), and mean concentrations of BCF constituents were compared with low risk (Na:K > 3, Type 2) and mixed cyst groups. Women with larger and more frequently occurring cysts had higher BCF estrogen and DHEA-S, and lower TGF-B1 levels. Women with Type 1 cysts had elevated BCF melatonin, estrogen, DHEA-S, and EGF, and lower concentrations of TGF-B2 compared to women with Type 2 cysts. BCF generally inhibited cell growth relative to serum-treated controls, consistent with previous studies. Melatonin and estrogen in BCF independently predicted growth inhibition and stimulation, respectively. Biological monitoring of BCF may help identify women with GCBD at greatest risk for breast cancer development.

Adult↗
PubMed1992

Relationships between androgen and estrogen sulfates in breast cyst fluid.

In 28 breast cyst fluids obtained from 20 patients (age 29-65 years) sodium, potassium and the sulfates (S) of estrone (E1), estradiol (E2), dehydroepiandrosterone (DHEA) and androsterone (A) were determined. The radioimmunoassays (RIA) used were validated for this particular biological fluid. According to electrolyte ratio (Na+/K+) the cyst fluids were subdivided into two groups: the first with low (less than 3) (n = 16) and the other with high (greater than 3) (n = 12) values. Markedly higher steroid sulfate levels were observed in the first group, the mean levels being: 147.7 nmol/l, 54.6 nmol/l, 108.1 mumol/l and 158.0 mumol/l for E1S, E2S, DHEAS and AS respectively. The mean levels in the second group were: 13.6 nmol/l, 6.7 nmol/l, 68.8 mumol/l and 33.6 mumol/l for E1S, E2S, DHEAS and AS, respectively. In the first group only E1S and E2S levels were significantly correlated (r = 0.51; P less than 0.05). Conversely, the steroid sulfate levels were significantly correlated with each other in the group with high electrolyte ratio. These data have confirmed preceding results and have clearly shown that breast cyst fluids with low electrolyte ratio contain more E2S than the other group. This finding might be correlated with the fact that patients with these breast cysts lined by with apocrine epithelium may be at a greater risk of breast cancer than those with the other type.

Adult↗
PubMed1998

Organochlorine compounds (DDE and PCB) in plasma and breast cyst fluid of women with benign breast disease.

The organochlorines, dichloro-diphenyl-trichloroethane and polychlorinated biphenyl (PCB) are pervasive environmental contaminants. Results from previous studies have been conflicting regarding the relationship between the internal dose of these organochlorine residues and breast cancer risk. To determine whether these compounds are present in breast cyst fluids and whether cyst fluid and plasma concentrations are correlated, we analyzed organochlorines in paired cyst fluid and plasma samples from 24 subjects using gas chromatography and electron capture detection. All but one of the women had a history of multiple cysts, suggesting that they were at elevated risk for future breast cancer. DDE (a metabolite of dichloro-diphenyl-trichloroethane) was present in 22 of the cyst samples and PCB was detected in 19 of the cyst samples. Organochlorine levels were more concentrated in the plasma than in breast cyst fluids. Levels of DDE in plasma were significantly correlated with those in cyst fluid (r = 0.73; P < 0.001); in contrast to PCB levels in cyst and plasma (r = 0.37; P = 0.12). Congener specific analysis of the PCBs showed that some individual congeners were preferentially excluded from or concentrated in the cyst fluid. To our knowledge, this study is the first to demonstrate that PCB and DDE are present in cyst fluids and thus in contact with the ductal epithelium of the breast. These results support the use of plasma DDE as a proxy for DDE in the target tissue in research on the role of environmental factors in breast cancer.

Adult↗
PubMed2003

Enterolactone in breast cyst fluid: correlation with EGF and breast cancer risk.

The purpose of our study was to investigate whether enterolactone does accumulate into breast cyst fluid and whether it correlates with breast cancer risk. We included 258 women who had at least one cyst aspiration and known intracystic cation and epidermal growth factor (EGF) concentration values. For 191 of such women serum aliquots were also available. The median value of serum enterolactone was 17 nM/l (range 1-140 nM/l). The median intracystic level of enterolactone was much higher (63 nM/l, range 0-872 nM/l) and was significantly higher in type I cysts (p = 0.000). This cyst type contained also significantly higher levels of EGF (p = 0.000). A direct relationship was found between serum and cyst fluid enterolactone levels (p = 0.000) and between cyst enterolactone and EGF levels (p = 0.03), the latter correlation being evident especially in type II cysts. Twelve patients in the cohort of women were found to have developed a breast cancer. After univariate analysis breast cancer risk was associated with cyst type and especially with EGF concentration. No association was evident for enterolactone concentration. However, enterolactone concentration appeared to significantly decrease the risk of patients with high EGF concentrations. Our results show that enterolactone does accumulate in breast cysts, and that it modulates the risk related to the intracystic level of EGF, which is confirmed to be a strong predictor of breast cancer risk.

4-Butyrolactone↗
PubMed1989

Relationship between epidermal growth factor and dehydroepiandrosterone and its sulphate in breast cyst fluid.

Gross cystic breast disease is a common condition. Women with apocrine breast cysts may be at higher risk of breast cancer than women with cysts which are lined by flattened epithelium. Apocrine cysts have been shown to be associated with higher intracystic levels of dehydroepiandrosterone sulphate and intracystic sodium to potassium ratios of less than 3. In this study we measured the concentrations of epidermal growth factor, dehydroepiandrosterone and its sulphate in breast cyst fluid. The concentrations of all three analytes were significantly higher in cysts with intracystic Na+/K+ ratios of less than 3 than in cysts with electrolyte ratios of greater than or equal to 3 (P less than 0.001). The higher levels of EGF in cysts with low intracystic electrolyte ratios may provide an explanation of why women with apocrine cysts may be at greater risk of breast cancer. Positive correlations were obtained between concentrations of EGF and DHAS and between EGF and DHA, compatible with the view that intracystic EGF levels may be androgen-modulated. A positive correlation was also obtained between DHA and DHAS concentrations which supports the view that DHA in cyst fluid may be derived from the metabolism of DHAS in the breast cyst wall.

Dehydroepiandrosterone↗
PubMedDate not supplied

Effect of breast cyst fluid on oestrone sulphatase activity in MCF-7 breast cancer cells.

Breast cyst fluid is a rich source of growth factors and sex hormones but the pathophysiology of cystic breast disease is largely unknown. In this study the net effects of breast cyst fluid on growth of, and oestrone sulphatase activity (oestrone sulphate --> oestrone) in the hormone-dependent MCF-7 and hormone-independent MDA-MB-231 breast cancer cell lines were assessed. Using a final breast cyst fluid dilution of 16.7% (v/v) MCF-7 cell growth was significantly inhibited by 7 of 21 samples (16% - 54%) while MDA-MB-231 cell growth was significantly inhibited by 17 of 20 samples (15% - 45%). Oestrone sulphatase activity was significantly inhibited in the MCF-7 cell line by 19 of 21 samples (22% - 81%) while significant stimulation of oestrone sulphatase activity was observed in MDA-MB-231 cells in 11 of 20 samples (17% - 149%). The presence of endogenous substances in breast cyst fluid which inhibit oestrone sulphatase activity in the MCF-7 cell line is exciting because of the therapeutic potential of oestrone sulphatase inhibition in hormone-dependent breast cancers.

Arylsulfatases↗
PubMed1988

Proteases in cyst fluid from human gross cyst breast disease.

Cyst fluid from women with gross cystic breast disease was found to contain protease activity when assayed against [14C]albumin. At least six different proteases were detected when the fluid was fractionated by a combination of S-300 Sephacel, hydroxylapatite, and DEAE-Sephacel chromatographic techniques. The distribution of the proteases appeared to be related to the ionic composition of the fluids. A major protease component, found in both high Na and high K fluids, was isolated. It showed chymotryptic cleavage characteristics against the beta-chain of insulin. It was partially inhibited by alpha 2-macroglobulin, N-tosyl-L-phenylalanine chloromethyl ketone, and benzamidine but not by leupeptin, pepstatin, N-tosyl-L-lysine chloromethyl ketone, or alpha 1-protease inhibitor. The protease has an apparent molecular weight of 110,000 with Mr 24,000 subunits. This protease may be identical or closely associated with Haagensen's GCDFP-24 progesterone binding protein which was isolated in a similar manner. An imbalance between protease and protease inhibitors in cyst fluid may account for gross cyst formation and may be involved in the tumorigenic process. The accumulation of poorly diffusible peptide fragments, as a result of protease activity, would increase the oncotic pressure leading to enlargement of the cyst cavity as water enters to reestablish osmotic equilibrium.

Female↗
PubMedDate not supplied

Transforming growth factor alpha, beta 1 and beta 2 in breast cyst fluid.

Women who have palpable breast cysts lined by apocrine metaplastic epithelium (intracystic Na/K < 3) may have a higher risk of breast cancer than those who have breast cysts lined by flattened epithelium (intracystic Na/K > 3). Transforming growth factor-alpha (TGF-alpha) exerts a proliferative effect on many cell types and may play a role in mammary carcinogenesis. Transforming growth factor-beta (TGF-beta) stimulates the proliferation of mesenchymal cells but inhibits mammary epithelial cell proliferation and may exert a protective effect against breast cancer development. The aim of this study was to measure the distribution of TGF-alpha, TGF-beta 1 and TGF beta 2 in the two sub-groups of breast cysts. TGF-alpha was undetectable in all but 2 of the 46 samples of breast cyst fluid tested. TGF-beta 1 ranged from undetectable up to 25.4 ng/ml (n = 46) while the intracystic concentrations of TGF-beta 2 were generally higher, ranging from 0.30 ng/ml to 125 ng/ml (n = 42). There was no significant difference between the distribution of TGF-beta 1 levels in the 2 cyst groups, but TGF-beta 2 levels were significantly higher (P < 0.001) in the high electrolyte ratio group (Na/K > 3). A negative correlation was found between TGF-beta 1 and TGF-beta 2 levels in the high electrolyte ratio group (rs = 0.605, p = 0.006, n = 19). TGF-alpha is unlikely to play an important role in breast cancer development in women with palpable breast cysts. The significantly higher concentrations of TGF-beta 2 in the high electrolyte ratio group may explain, in part, the lower risk of breast cancer in this group of women.

Biopsy, Needle↗
PubMed1992

A case control study of factors associated with macroscopic breast cysts.

Women who have a breast cyst aspirated are at increased risk of developing breast cancer. We present an age-matched case control study comparing the reproductive characteristics of 352 women who had a breast cyst aspirated with a control group of 352 contemporaneous clinically normal women. Women with breast cysts were more likely to be nulliparous [odds ratio (OR) = 2.28, 95% confidence interval (CI) = 1.34-3.88] or have a late age at first live birth (chi 2 trend = 5.6, P less than 0.025), and a late menopause (chi 2 trend = 4.3, P less than 0.05). They were less likely to have ever used the oral contraceptive pill (OR = 0.38, 95% CI = 0.26-0.55) or to have used the pill for a short duration (chi 2 trend = 16.8, P less than 0.001), and were less likely to have had a hysterectomy (OR = 0.58, 95% CI = 0.36-0.93). They were more likely to wear a small bra (chi 2 trend = 18.6, P less than 0.001) and bra cup (chi 2 trend = 5.6, P less than 0.025). Nulliparity, late age at first live birth and late menopause are factors common to breast cancer risk.

Adult↗
PubMed1991

Accumulation of active androgens in breast cyst fluids.

80 breast cyst fluids (BCF) from 57 patients were divided by K+/Na+ ratio: 56 with ratio over 1 (type I) and 24 with ratio less than 1 (type II). Significantly higher amounts of testosterone, dihydrotestosterone and dehydroepiandrosterone sulphate (DHAS) were found in type I than in type II cysts. A positive relation was found between testosterone and dihydrotestosterone in both types. DHAS was significantly correlated with testosterone and dihydrotestosterone in type I casts only. In 52 patients, blood was sampled after cyst evacuation. Testosterone was significantly higher in blood than in BCF while dihydrotestosterone and androstenedione were significantly higher in BCF. No relation was observed between circulating levels of androgens and their intracystic concentrations. Women bearing type I cysts may be at increased risk of developing cancer. These findings support the hypothesis that androgens play a role in the hormonal aetiology of breast cancer.

Adult↗
PubMed1995

Relationship between basic fibroblast growth factor and transforming growth factor-beta 2 in breast cyst fluid.

Women with "apocrine" breast cysts (usually having intracystic Na/K < 3) may have a higher risk of developing breast cancer than women with breast cysts lined by flattened epithelium (usually having intracystic Na/K > 3). In this study the concentrations of basic fibroblast growth factor (bFGF), a potent mitogen, and transforming growth factor-beta 2 (TGF-beta 2), which exerts a growth inhibitory effect on epithelial cell types, were measured in breast cyst fluid and their relationship studied. Both growth factors were measured by "sandwich" enzyme immunometric assays. The concentrations of both bFGF and TGF-beta 2 were significantly higher (P < 0.001) in the Na/K > 3 group (median 444 fmol/L, range: < 56 fmol/L-7,890 fmol/L, n = 23 and median 1,776 pmol/L, range: 20.4 pmol/L-5,000 pmol/L, n = 19 respectively) than in the Na/K < 3 group (median < 56 fmol/L, range: < 56 fmol/L-2,722 fmol/L, n = 21 and median 176 pmol/L, range: 12 pmol/L-1,940 pmol/L, n = 23 respectively). Significantly positive correlations were found between bFGF and TGF-beta 2 (rS = 0.496, n = 37, P = 0.002), bFGF and Na/K (rS = 0.599, n = 44, P < 0.001) and TGF-beta 2 and Na/K (rS = 0.521, n = 42, P < 0.001). The significantly higher concentrations of the growth inhibitory TGF-beta 2 in the Na/K > 3 cyst group may provide an explanation for the lower risk of breast cancer which has been observed in this group of women. The role of bFGF in mammary carcinogenesis is unclear as lower levels of this growth factor are present in breast cancer tissue and breast cancer cell lines than in normal breast tissue and cell lines. The positive correlation between bFGF and TGF-beta 2 may indicate regulation by a common factor or that one of these growth factors may regulate the production of the other. This is the subject of further study.

Electrolytes↗
PubMed2006

Breast cyst fluids increase the proliferation of breast cell lines in correlation with their hormone and growth factor concentration.

OBJECTIVE AND DESIGN: Gross cystic disease (GCD) of the breast is reported to occur in 7% of women in the developed world and, although not premalignant, is thought to be associated with an increased risk of breast cancer. Hormone and growth factor concentration levels were measured in breast cyst fluid (BCF) to correlate them with their mitogenic activity in tumour (MCF-7) or nontransformed (MCF-10A) cells. RESULTS: Oestradiol (E2), oestrone (E1), E2-sulfate (E2-S), E1-sulfate (E1-S) and epidermal growth factor (EGF) concentrations were, as expected, significantly higher in type I than in type II cysts, while transforming growth factor-beta 2 (TGF-beta2) showed higher levels in type II cysts. Fifty per cent of the BCF samples stimulated [3H]-thymidine incorporation into MCF-7 cells while 34.5% inhibited this parameter. In MCF-10A cells, most BCF samples were stimulatory (85%). E2, E1 and EGF concentrations in BCF samples correlated significantly and positively with cell proliferation in MCF-7 cells, whereas a significant negative correlation was found for TGF-beta2. In MCF-10A cells, only E2-S and E1-S exhibited significant positive correlation, whereas a significant negative correlation was found for TGF-beta2. Progesterone (Pg), E2 and EGF incubated under the same conditions had a stimulatory effect on [3H]-thymidine incorporation into MCF-7 cells, whereas TGF-beta2 inhibited this parameter. Pg, E2, E1 and EGF significantly stimulated this parameter in MCF-10A cells. CONCLUSIONS: The stimulatory action of BCF on cell proliferation in a model of human breast epithelial cells could partly explain the increased incidence of breast cancer in cyst-bearing women.

Adult↗
PubMedDate not supplied

Oestradiol and epidermal growth factor in breast cyst fluid.

Gross cystic breast disease may be categorised into 2 groups according to the intracystic cation ratios (Na+/K+ less than 3 and Na+/K+ greater than 3). It has been suggested that women in the low electrolyte ratio group may be at higher risk of developing breast cancer. Epidermal growth factor concentrations in breast cyst fluid were significantly higher in the low than in the high electrolyte ratio group but there was no significant difference in oestradiol concentrations between the two groups of breast cysts. A negative correlation was obtained between intracystic concentrations of epidermal growth factor and Na+/K+ (rs = -0.617, p less than 0.001). No evidence was found to support the hypothesis that concentrations of epidermal growth factor may be modulated by oestradiol. The higher concentrations of epidermal growth factor in the low electrolyte ratio group may provide an explanation for the higher risk of breast cancer observed in this group.

Biomarkers↗