Epidermal growth factor receptors in breast cancer: association with early relapse and death, poor response to hormones and interactions with neu

Epidermal growth factor receptors (EGFRs) were measured in 221 primary breast cancers by ligand binding with 125I-labelled EGF, and high-affinity sites were quantitated. There was a highly significant inverse relationship between oestrogen receptor (ER) and EGFR (15 EGFR-positive [EGFR+]ER+ and 92 EGFR-negative [EGFR-]ER+: 54 EGFR- ER- and 60 EGFR+ ER-). The relapse-free survival and overall survival were significantly shorter for EGFR+ vs EGFR- tumours (P less than 0.001) by about 2 yr in the case of relapse-free survival. When ER- tumours were substratified by EGFR status, the EGFR- ER- tumours had a prognosis almost as good as the ER+ tumours. In 31 of 184 cases, high expression of neu, correlating with amplification, was found. Expression of neu conferred similar poor prognosis to EGFR expression in all prognostic subgroups. Coexpression of neu and EGFR had an additive adverse effect. Epidermal growth factor receptors (EGFR) and oestrogen receptors (ER) were analysed in 221 patients with primary operable breast cancer by means of radioligand assays. After median follow-up of 24 months (range 3-60 months), there had been recurrences in 99 patients, of whom 72 (median age 56 yr, range 32-77 yr) received tamoxifen alone as first-line treatment for recurrence. 14 patients (19%) showed a response to this therapy and 58 (81%) did not. Of 32 ER+ tumours, 12 (37.5%) showed an objective response to tamoxifen compared with only 2 of 40 (5%) ER- tumours (P less than 0.005). Of 35 EGFR+ tumours, 3 (8.5%) achieved an objective response compared with 11 of 36 (30%) EGFR tumours (P less than 0.05). Only 1 of 28 EGFR+, ER- tumours achieved an objective response. Including patients whose disease remained stable for more than 6 months with the responders, however, EGFR status was a better predictor of response to tamoxifen; 15 of 37 EGFR- patients and 5 of 35 EGFR+ patients responded (P less than 0.01).     

Estrogen receptor and C19-5-ene-steroid concentrations in the nuclear fraction from human breast carcinoma tissue.

The adrenal-derived estrogen 5-androstene-3 beta,17 beta-diol (ADIOL) is estrogenic at the concentrations found in the blood of Western women. We have now measured the concentrations of both ADIOL and the estrogen receptor (ER) in the nuclear fraction (800 g pellet) of 89 primary human mammary tumors. No difference was found in nuclear ADIOL concentrations in tumors from 45 pre- and 44 postmenopausal women. Significantly higher nuclear ADIOL concentrations were found in 49 ER negative tumors compared to 40 ER positive tumors (P < 0.005). A similar relationship applied in the postmenopausal group (P = 0.01) and the premenopausal group, but in this latter instance failed to reach significance (P = 0.1). In ER positive tumors there was no correlation between ADIOL and ER nuclear levels. ADIOL was present in the total particulate fraction (100,000 g pellet) at twice the concentration found in the nuclear 800 g pellet and again no difference was found in its concentration in tumors from 20 pre- compared to 34 postmenopausal women. Dehydroepiandrosterone was also measured in the 800 g fraction of 45 tumors and its concentration, which was some 10-fold higher than ADIOL and significantly correlated with that steroid, was again independent of menopausal status. The higher concentration of C19-5-ene-steroids in ER negative cellular fractions could be due to differences in their metabolism; ER negative tumors either lack, or possess very low levels of, hydroxysteroid sulfotransferase which catalyzes formation of sulfate esters of C19-5-ene-steroids previously observed to be major metabolites produced by ER positive cells. Higher concentrations of free steroids in ER negative cells would then be available for combination with membranes and non-specific binding sites throughout the cell.     

Patterns of estrogen and progesterone receptors in rhesus monkey endometrium during secretory phase of normal menstrual cycle and preimplantation stages of gestation

Using validated methods, estradiol receptor (ER) and progesterone receptor (PgR) levels have been estimated in endometria collected in secretory phase of normal menstrual cycle and preimplantation stages of gestation from rhesus monkeys (Macaca mulatta). Endometrial PgR in both cytoplasmic and nuclear compartments decreased significantly (P less than 0.001) from day 2 to day 6 post-ovulation in both groups, but in fertile cycle, absolute levels of nuclear PgR remained significantly higher (P less than 0.05) on days 4, 5 and 6 of gestation, ER concentrations, both total (P less than 0.02), as well as cytoplasmic (P less than 0.01) declined significantly in secretory phase of normal menstrual cycle while nuclear ER levels remained unchanged. In the preimplantation period, ER patterns remained unvarying on days 2-6 of gestation in both cytoplasmic and nuclear compartments; their levels in nuclear fraction were significantly higher from day 3 onwards while, total cytoplasmic ER concentrations were higher from day 4 of gestation compared with the values obtained for secretory phase tissues from normal ovulatory cycles. No changes were, however, detected in apparent equilibrium dissociation constants (Kd) for the sex steroid receptors in endometria obtained from fertile and non-fertile cycles. It has been suggested that in prenidatory stage rhesus monkey endometrium elevated concentrations of nuclear ER and PgR possibly indicate higher degree of nuclear occupancy required for endometrial differentiation permitting blastocyst implantation.     

S100A14在乳腺癌不同分子亚型中表达的临床病理研究

目的:探讨S100钙结合蛋白A14(S100A14)在乳腺癌不同分子亚型中的表达及临床病理意义,为确定新的分子分型标志物提供参考依据。方法:254例乳腺癌石蜡组织来源于2013年1月16日至2014年5月22日在中南大学湘雅医学院附属肿瘤医院暨湖南省肿瘤医院进行乳腺癌根治术的患者。应用免疫组织化学方法检测S100A14在乳腺癌组织中的表达,分析其S100A14在不同分子亚型乳腺癌组织中表达及其与患者临床病理指标间的相关性,采用Kaplan-Meier法分析S100A14蛋白表达与乳腺癌患者预后的关系。结果:S100A14在ER+/PR+/HER2+型、ER+/PR+/HER2-型、ER-/PR-/HER2+型、ER-/PR-/HER2-型乳腺癌四种分子亚型中的阳性表达分别为38.5%、47.1%、75.5%、80.0%,以在ER-/PR-/HER2-型中表达最高,在ER+/PR+/HER2+型中表达最低,四组间的阳性表达比较差异有显著统计学意义(P0.01);S100A14的表达与乳腺癌患者术后肝转移呈正相关(r=0.134,P0.05),与ER、PR表达均呈负相关(r=-0.353,P0.01),而与ER+/PR+/HER2+型、ER+/PR+/HER2-型乳腺癌的临床病理特征无显著相关性(P0.05)。在ER-/PR-/HER2+型乳腺癌中,有腋窝淋巴结转移组患者的S100A14阳性表达率明显高于无腋窝淋巴结转移组,差异有统计学意义(P0.05);在ER-/PR-/HER2-型中,S100A14表达与术后肺转移呈负相关(r=-0.272, P=0.044)。结论:S100A14在不同分子亚型乳腺癌中表达存在差异,其表达与不同分子类型乳腺癌转移或复发有关,可能作为乳腺癌分子分型的候选标记物。     

Epidermal growth factor receptor (EGFr) as a marker for poor prognosis in node-negative breast cancer patients: Neu and tamoxifen failure

Analysis of EGFr and ER was performed on tumour samples from 231 patients with operable breast cancer followed for up to 6 yr after surgery. The median duration of follow-up in patients still alive at the time of analysis was 45 months.

Thirty-five percent of patients (82) had tumours greater than 10 fmol/mg ¹²⁵I-EGF binding (EGFr⁺) and 47% (109) had cystolic ER concentration >5 fmol/mg (ER⁺), with a marked inverse relationship between EGFr and ER (P<0.00001). EGFr was second only to axillary node status as a prognostic marker for all patients both in terms of relapse-free and overall survival (P<0.001, logrank EGFr⁺ vs EGFr⁻).

For patients with histologically negative axillary nodes EGFr was superior to ER in predicting relapse and survival (P<0.01 and P<0.005, respectively, compared to P<0.1 and P<0.1, logrank). In a multivariate (Cox model) analysis only EGFr, out of EGFr, ER, size and grade, was predictive for either relapse-free or overall survival for patients with node-negative disease (P=0.052 and P=0.026, respectively).

The correlation of neu expression with response to tamoxifen in patients with recurrent disease was assessed immunochemically. Response rate was reduced in the presence of neu from 50 to 17% for ER⁺ cases and from 26 to 0% for ER⁻ cases.     

Ca2+-dependent facilitated shortening in isotonic contraction of trachealis muscle

We compared isotonic shortening with isometric force generation as a function of external Ca2+ in 166 tracheal smooth muscle (TSM) strips from 27 mongrel dogs in vitro. Concentration-response curves were generated with muscarinic stimulation (acetylcholine, ACh), alpha-adrenergic receptor activation (norepinephrine after beta-adrenoceptor blockade, NE), serotonin (5-HT), and KCl-substituted Krebs-Henseleit solution. The concentrations of 5-HT causing half-maximal shortening (ECS50, 1.54 +/- 0.14 X 10(-7) M) and half-maximal active isometric tension (ECT50, 1.72 +/- 0.30 X 10(-7) M) were similar (P = NS). Likewise, ECS50 (21.9 +/- 0.7 mM) and ECT50, (22.0 +/- 0.9 mM) were similar for KCl. In contrast, facilitated isotonic shortening (i.e., greater isotonic shortening for comparable degrees of force generation) was elicited with ACh and NE for all levels of force generation between 15 and 85% of maximum and for all concentrations of ACh from 3 X 10(-8) to 3 X 10(-5) M (P less than 0.05 for all points). Facilitated isotonic shortening also was elicited for all concentrations of NE from 10(-8) to 10(-6) M (P less than 0.05 for all points). Removal of Ca2+ from the perfusate substantially reduced the potency of ACh (P less than 0.001) and abolished differences between ECS50 (2.23 +/- 0.28 X 10(-5) M) and ECT50 (2.50 +/- 0.46 X 10(-5) M, P = NS). We demonstrate that for comparable degrees of force generation, muscarinic and alpha-adrenergic receptor activation cause greater isotonic shortening than KCl or 5-HT and that this facilitated shortening is associated with the concentration of external Ca2+.     

Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines.

Lack of estrogen receptor (ER) and presence of vimentin (VIM) associate with poor prognosis in human breast cancer. We have explored the relationships between ER, VIM, and invasiveness in human breast cancer cell lines. In the matrigel outgrowth assay, ER+/VIM- (MCF-7, T47D, ZR-75-1), and ER-/VIM- (MDA-MB-468, SK-Br-3) cell lines were uninvasive, while ER-/VIM+ (BT549, MDA-MB-231, MDA-MB-435, MDA-MB-436, Hs578T) lines formed invasive, penetrating colonies. Similarly, ER-/VIM+ cell lines were significantly more invasive than either the ER+/VIM- or ER-/VIM- cell lines in the Boyden chamber chemoinvasion assay. Invasive activity in nude mice was only seen with ER-/VIM+ cell lines MDA-MB-231, MDA-MB-435 and MDA-MB-436. Hs578T cells (ER-/VIM+) showed hematogenous dissemination to the lungs in one of five mice, but lacked local invasion. The ER-/VIM+ MCF-7ADR subline was significantly more active than the MCF-7 cells in vitro, but resembled the wild-type MCF-7 parent in in vivo activity. Data from these cell lines suggest that human breast cancer progression results first in the loss of ER, and subsequently in VIM acquisition, the latter being associated with increased metastatic potential through enhanced invasiveness. The MCF-7ADR data provide evidence that this transition can occur in human breast cancer cells. Vimentin expression may provide useful insights into mechanisms of invasion and/or breast cancer cell progression.     

Loss of circadian clock gene expression is associated with tumor progression in breast cancer

Several studies suggest a link between circadian rhythm disturbances and tumorigenesis. However, the association between circadian clock genes and prognosis in breast cancer has not been systematically studied. Therefore, we examined the expression of 17 clock components in tumors from 766 node-negative breast cancer patients that were untreated in both neoadjuvant and adjuvant settings. In addition, their association with metastasis-free survival (MFS) and correlation to clinicopathological parameters were investigated. Aiming to estimate functionality of the clockwork, we studied clock gene expression relationships by correlation analysis. Higher expression of several clock genes (e.g., CLOCK, PER1, PER2, PER3, CRY2, NPAS2 and RORC) was found to be associated with longer MFS in univariate Cox regression analyses (HR<1 and FDR-adjusted P < 0.05). Stratification according to molecular subtype revealed prognostic relevance for PER1, PER3, CRY2 and NFIL3 in the ER+/HER2- subgroup, CLOCK and NPAS2 in the ER-/HER2- subtype, and ARNTL2 in HER2+ breast cancer. In the multivariate Cox model, only PER3 (HR = 0.66; P = 0.016) and RORC (HR = 0.42; P = 0.003) were found to be associated with survival outcome independent of established clinicopathological parameters. Pairwise correlations between functionally-related clock genes (e.g., PER2-PER3 and CRY2-PER3) were stronger in ER+, HER2- and low-grade carcinomas; whereas, weaker correlation coefficients were observed in ER- and HER2+ tumors, high-grade tumors and tumors that progressed to metastatic disease. In conclusion, loss of clock genes is associated with worse prognosis in breast cancer. Coordinated co-expression of clock genes, indicative of a functional circadian clock, is maintained in ER+, HER2-, low grade and non-metastasizing tumors but is compromised in more aggressive carcinomas.     

Estrogen-independent effects of ER-α36 in ER-negative breast cancer

Estrogen receptor-alpha 36 (ER-α36) is a variant of ER-α that has been found to be expressed in conventional ER (ER-α66)-negative breast cancer cell lines and human breast cancer samples. In this study, we found that, using immunohistochemical study, ER-α36 expression was significantly higher in ER-negative tumors than in ER-positive tumors although the expression was not associated with other clinicopathological characteristics. We then constructed an ER-α36-specific microRNA hairpin vector and established stable ER-α36 knockdown cells, and found that the knockdown cells were more sensitive to paclitaxel; the c-Jun N-terminal kinase pathway appeared to be involved in the mechanism. Downregulation of ER-α36 also resulted in decreased migration and invasion. These changes were estrogen independent. Our findings indicated that target ER-α36 may be a strategy for treating ER-negative breast cancers.     

Tissue microarray study for classification of breast tumors

Clinical and pathological heterogeneity of breast cancer hinders selection of appropriate treatment for individual cases. Molecular profiling at gene or protein levels may elucidate the biological variance of tumors and provide a new classification system that correlates better with biological, clinical and prognostic parameters. We studied the immunohistochemical profile of a panel of seven important biomarkers using tumor tissue arrays. The tumor samples were then classified with a monothetic (binary variables) clustering algorithm. Two distinct groups of tumors are characterized by the estrogen receptor (ER) status and tumor grade (p = 0.0026). Four biomarkers, c-erbB2, Cox-2, p53 and VEGF, were significantly overexpressed in tumors with the ER-negative (ER-) phenotype. Eight subsets of tumors were further identified according to the expression status of VEGF, c-erbB2 and p53. The malignant potential of the ER-/VEGF+ subgroup was associated with the strong correlations of Cox-2 and c-erbB2 with VEGF. Our results indicate that this molecular classification system, based on the statistical analysis of immunohistochemical profiling, is a useful approach for tumor grouping. Some of these subgroups have a relative genetic homogeneity that may allow further study of specific genetically-controlled metabolic pathways. This approach may hold great promise in rationalizing the application of different therapeutic strategies for different subgroups of breast tumors.     

Salivary IgA subclasses and infection risk in elite swimmers.

The concentrations of total IgA, IgA1 and IgA2 were measured in saliva collected from 25 elite swimmers in the early and late phases of a 7 month training season and compared with the number of respiratory infections during the season. The IgA1 concentrations in the early phase of the training season were significantly associated (P = 0.01) with the number of respiratory infection episodes during the training season. The lower the concentration of IgA1, the greater the number of infection episodes. Swimmers with four or more infections during the training season had significantly lower salivary IgA1 concentrations than those with less than four infection episodes (P = 0.01). The proportion of IgA1 in the saliva of the elite swimmers (80%) was higher than for normal non-exercising adults (60%). A small proportion of athletes had salivary IgA2 concentrations below the detection limit of the assay and the mean concentration of IgA2 was significantly lower than the concentrations for a normal adult population (P = 0.01). This study suggests that measurement of IgA subclasses, in particular IgA1, at the commencement of a training season may predict infection risk in elite swimmers.     

Effect of antigravity suit inflation on cardiovascular, PRA, and PVP responses in humans

Blood pressure, pulse rate (PR), serum osmolality and electrolytes, as well as plasma vasopressin (PVP) and plasma renin activity (PRA), were measured in five men and two women [mean age 38.6 +/- 3.9 (SE) yr] before, during, and after inflation of an antigravity suit that covered the legs and abdomen. After 24 h of fluid deprivation the subjects stood quietly for 3 h: the 1st h without inflation, the 2nd with inflation to 60 Torr, and the 3rd without inflation. A similar control noninflation experiment was conducted 10 mo after the inflation experiment using five of the seven subjects except that the suit was not inflated during the 3-h period. Mean arterial pressure increased by 14 +/- 4 (SE) Torr (P less than 0.05) with inflation and decreased by 15 +/- 5 Torr (P less than 0.05) after deflation. Pulse pressure (PP) increased by 7 +/- 2 Torr (P less than 0.05) with inflation and PR decreased by 11 +/- 5 beats/min (P less than 0.05); PP and PR returned to preinflation levels after deflation. Plasma volume decreased by 6.1 +/- 1.5% and 5.3 +/- 1.6% (P less than 0.05) during hours 1 and 3, respectively, and returned to base line during inflation. Inflation decreased PVP from 6.8 +/- 1.1 to 5.6 +/- 1.4 pg/ml (P less than 0.05) and abolished the significant rise in PRA during hour 1. Both PVP and PRA increased significantly after deflation: delta = 18.0 +/- 5.1 pg/ml and 4.34 +/- 1.71 ng angiotensin I X ml-1 X h-1, respectively. Serum osmolality and Na+ and K+ concentrations were unchanged during the 3 h of standing.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sweat lactate secretion during exercise in relation to women's aerobic capacity

The purpose of this investigation was to determine whether sweat lactate secretion during exercise [approximately 70% maximum O2 consumption (VO2max), 60 min] differed in active vs. sedentary female subjects. Sweat rate, total sweat lactate secretion, and sweat lactate concentration were monitored in a group of sedentary (VO2max = 41.0 +/- 1.62 ml X kg-1 X min-1) and active (VO2max = 51.2 +/- 3.20 ml X kg-1 X min-1) women. Sweat rate was significantly (P less than 0.05) greater in the active subjects. There was a significant difference between groups in total amount of sweat lactate secreted (P less than 0.05), with the active group secreting less lactate (29.8 +/- 5.03 mmol, mean +/- SE) than the sedentary group (50.2 +/- 6.61 mmol). Concomitant with the lower total sweat lactate secretion in the active subjects was a significantly (P less than 0.05) more dilute sweat lactate concentration (42.6 +/- 14.08 vs. 100.4 +/- 32.37 mM). In these female subjects, sweat lactate concentration was inversely correlated (r = -0.79, P less than 0.01, n = 10) to sweat rate. It is concluded that total sweat lactate loss is significantly less in active than in sedentary women and that the active subjects secrete a greater quantity of lactate dilute sweat.     

The nuclear DNA content of human breast carcinoma. Associations with clinical stage, axillary lymph node status, estrogen receptor status and outcome

Using Feulgen-DNA cytophotometry, the nuclear DNA content was determined in specimens from 169 female patients with unilateral primary carcinoma of the breast. The tumors were classified as either diploid (73 cases: 43%) or hyperdiploid (96 cases), according to the ploidy of the tumor cells. Statistically significant associations were found between the DNA content and other characteristics of the patients and their tumors. (1) In postmenopausal women, inoperable tumors were more likely to be hyperdiploid (P less than .005). (2) In patients with operable disease, diploid tumors were less likely to have metastasized to the axillary lymph nodes (P less than .005) and were also less likely to have four or more positive nodes (P = .0044). (3) Overall, 71% of the diploid tumors and 52% of the hyperdiploid tumors were estrogen-receptor (ER) positive. This difference in proportions was statistically significant (P less than .05), but when the patients were divided into premenopausal and post-menopausal groups, the proportions of ER-positive tumors were not significantly different in either group. (4) In 113 patients considered suitable for studies on outcome (mean length of follow-up of 27 months, with a range from 0 to 71 months), the rates of relapse were 3 of 55 diploid cases and 17 of 58 hyperdiploid cases. The rate of relapse was higher in the hyperdiploid group, irrespective of lymph node status.     

Estrogen Receptor Expression Is Associated with DNA Repair Capacity in Breast Cancer

Estrogen-receptor-positive (ER+) tumors employ complex signaling that engages in crosstalk with multiple pathways through genomic and non-genomic regulation. A greater understanding of these pathways is important for developing improved biomarkers that can better determine treatment choices, risk of recurrence and cancer progression. Deficiencies in DNA repair capacity (DRC) is a hallmark of breast cancer (BC); therefore, in this work we tested whether ER signaling influences DRC. We analyzed the association between ER positivity (% receptor activation) and DRC in 270 BC patients, then further stratified our analysis by HER2 receptor status. Our results show that among HER2 negative, the likelihood of having low DRC values among ER- women is 1.92 (95% CI: 1.03, 3.57) times the likelihood of having low DRC values among ER+ women, even adjusting for different potential confounders (p<0.05); however, a contrary pattern was observed among HER2 positives women. In conclusion, there is an association between DRC levels and ER status, and this association is modified by HER2 receptor status. Adding a DNA repair capacity test to hormone receptor testing may provide new information on defective DNA repair phenotypes, which could better stratify BC patients who have ER+ tumors. ER+/HER2- tumors are heterogeneous, incompletely defined, and clinically challenging to treat; the addition of a DRC test could better characterize and classify these patients as well as help clinicians select optimal therapies, which could improve outcomes and reduce recurrences.     

Somatomedin levels in the diagnosis and therapy of growth hormone deficiency

Serum somatomedin-C (SM-C) and somatomedin (SM) concentrations were measured by, respectively, radioimmuno (SM-C RIA) and radioreceptor assays (SM RRA) in 3 groups of children with short stature. The patient population was different from previously reported series in that it was urban Brazilian, low income, and significantly older. Group A consisted of 6 male and 3 female children, aged 7.7-16.0 years, whose average peak plasma immunoreactive growth hormone (GH) was above 10 ng/ml. Group B contained 8 male and 5 female untreated GH-deficient patients, ranging in age from 9.5 to 21.0 years. In Group C there were 4 male and 1 female GH-deficient subjects treated with I.M. injections of GH (0.1 U/kg) from 1 month to 7 years. The mean +/- SE basal RIA SM-C (ng/ml) concentrations were significantly lower in groups B (34.2 +/- 8.8) and C (43.8 +/- 13.7) than A (214.3 +/- 42.7): A X B, P less than 0.001 and A X C, P less than 0.02. Likewise the mean +/- SE basal RRA SM (ng/ml) concentrations were significantly lower in groups B (78.9 +/- 17.6) and C (90.8 +/- 19.3) than group A (316.3 +/- 43.0): A X B, P less than 0.001 and A X C, P less than 0.002. A significant linear correlation was observed between RIA and RRA in group B (r = 0.84; P less than 0.001) and C (r = 0.96; P less than 0.01), but not for A (r = 0.61; P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)