Quantitative Measurement of Serum MicroRNA-21 Expression in Relation to Breast Cancer Metastasis in Chinese Females

BACKGROUND: Breast cancer is the most common type of cancer in females. Aberrant expression of microRNA-21 (miR-21) has previously been reported in breast cancer tissue. The aim of this study was to investigate expression levels of serum miR-21 in breast cancer patients and evaluate its prognostic value in Chinese females. METHODS: Real-time quantitative (RQ)-PCR was used to analyze miR-21 expression in archived serum, tumor tissue, and adjacent normal tissue from 549 participants (326 with breast cancer, 223 without breast cancer). We also analyzed associations between serum miR-21 expression and breast cancer subtypes and patient prognosis. Recurrence and survival were analyzed by using the multivariate Cox proportional hazards model. RESULTS: Expression of miR-21 was significantly higher in breast cancer tissues compared with normal adjacent breast tissues (P<0.001). The 2(-DeltaDeltaCt) values for serum miR-21 in breast cancer patients versus healthy controls were 9.12+/-3.43 and 2.96+/-0.73, respectively. Multivariate Cox proportional hazards model suggested that serum miR-21 expression was an independent poor prognostic factor for both recurrence (hazard ratio [HR]= 2.942; 95% confidence interval [CI]=1.420-8.325; P=0.008) and disease-free survival (HR=2.732; 95% CI=1.038-7.273, P=0.003) in breast cancer. CONCLUSIONS: Increased serum miR-21 expression level was correlated with poor prognosis of breast cancer patients, indicating that serum miR-21 may be a novel prognostic marker for recurrence and survival of breast cancer patients before resection.

Comparison of Methylation Profiling in Cancerous and Their Corresponding Normal Tissues from Korean Patients with Breast Cancer

BACKGROUND: Aberrant DNA hypermethylation plays a pivotal role in carcinogenesis and disease progression; therefore, accurate measurement of differential gene methylation patterns among many genes is likely to reveal biomarkers for improved risk assessment. We evaluated the gene hypermethylation profiles of primary breast tumors and their corresponding normal tissues and investigated the association between major clinicopathological features and gene hypermethylation. METHODS: A single reaction using methylation-specific multiplex ligation-dependent probe amplification was used to analyze the DNA methylation status of 24 tumor suppressor genes in 60 cancerous tissues and their corresponding normal tissues from patients with primary breast cancer. RESULTS: In cancerous breast tissues, 21 of 24 genes displayed promoter methylation in one or more samples. The most frequently methylated genes included RASSF1 (43.3%), APC (31.7%), CDKN2B (25.0%), CDH13 (23.3%), GSTP1 (16.7%), and BRCA1 (10%). APC was associated with lymph node metastasis, and BRCA1 was associated with negative estrogen receptor and negative progesterone receptor expression. In normal breast tissues, 8 of 24 tumor suppressor genes displayed promoter hypermethylation; CDKN2B (28.3%) and RASSF1 (8.3%) hypermethylation were most frequently observed. CONCLUSIONS: RASSF1 and CDKN2B hypermethylation in Korean breast cancer patients were the most frequent in cancerous tissue and corresponding normal tissue, respectively. Our data indicates that methylation of specific genes is a frequent event in morphologically normal breast tissues adjacent to breast tumors as well as the corresponding breast cancers. This study also suggests that gene methylation is linked to various pathological features of breast cancer; however, this requires confirmation in a larger study.

Diffuse Intense 18F-FDG Uptake at PET in Unilateral Breast Related to Breastfeeding Practice

We present an interesting case of incidental diffuse fluorodeoxyglucose (FDG) uptake at PET in her left breast, related to atypical breastfeeding practice. Clinically, differential diagnoses of diffuse intense FDG uptake in unilateral breast include advanced breast cancer, breast lymphoma and inflammatory condition. However, normal physiologic lactation may also show increased FDG uptake in the breasts. Therefore, if we encounter that finding in daily practice, we should question the patient regarding unilateral breastfeeding. In addition, mammography and ultrasound would be helpful to confirm the diagnosis.

La relación núcleo-citoplasmática en la diferenciación y transformación celular del epitelio mamario / The nucleo-cytoplasmic relationship in the differentiation and cellular transformation of the mammary epithelium

ANTECEDENTES: Las células pertenecientes a la línea HC11 son estimuladas tanto a proliferar mediante el Factor de Crecimiento Epidérmico, como a diferenciar estimuladas con dexametasona, insulina y prolactina, dando origen a los tipos celulares HC11 GM y HC11 IM, respectivamente. De igual manera cuando ellas son transfectadas con el oncogén ras generan Q6 GM y Q6 IM, células transformadas. Producto de ambos mecanismos estas células asumen distintas propiedades modificando tanto sus componentes como sus funciones, los cuales pueden ser cuantificados mediante técnicas morfométricas. OBJETIVO: Evidenciar en términos cuantitativos y morfológicos las variaciones experimentadas tanto por los núcleos como los citoplasmas y su correspondiente relación núcleo-citoplasmática (N/C) pertenecientes a células mamarias de la línea HC11 con el decorrer de los mecanismos de diferenciación y de transformación celular. MÉTODO: Se estudió a nivel de microscopia electrónica de transmisión los tipos celulares en proceso de diferenciación y transformación, cuantificando variaciones de la relación núcleo-citoplasmática y su respectiva funcionalidad. RESULTADOS: Se evidencian diferencias estadísticamente significativas referentes a las áreas nucleares y citoplasmáticas pertenecientes a estos tipos celulares. CONCLUSIÓN: Las células del epitelio mamario en proceso de diferenciación como de transformación, presentan diferentes valores en su relación N/C hecho que responde a funcionalidades específicas en cada tipo celular.

BACKGROUND: Cells of the HC11 line are stimulated to proliferate using the Epidermic Growth Factor, and to differentiate with dexamethasone, insulin and prolactin, giving rise to cell types HC11 GM and HC11 IM, respectively. Likewise when they are transfected with the ras oncogen they generate Q6 GM and Q6 IM transformed cells. As a result of these two mechanisms, these cells assume different properties, in which both their components and their functions are modified. The modifications can be quantified by morphometric techniques. OBJECTIVE: To show in quantitative and morphological terms the variations effected in both the nuclei and the cytoplasms, and the corresponding nuclear-cytoplasmic ratio of mammary cells of the HC11 line, under the effects of cellular differentiation and transformation mechanisms. METHOD: The cell types undergoing differentiation and transformation processes were studied by transmission electron microscope, permitting quantification of variations in the nuclear-cytoplasmic ratio and its relation with the respective functions. RESULTS: Statistically significant differences were found in the nuclear and cytoplasmic areas of these cell types. CONCLUSION: cells of the mammary epithelium undergoing differentiation and transformation processes present different values for their nuclear-cytoplasmic ratio, and this responds specific functions in each cell-type.

Ação da tibolona sobre o parênquima mamário / Action of tibolone on the mammary parenchyma

Tibolona é uma substância hormonal, com ação tecido-específica, usada no tratamento de sintomas do climatério e na prevenção de osteoporose em mulheres na pós-menopausa. Esse trabalho objetivou revisar os efeitos da tibolona no parênquima mamário. Sob ação da tibolona, as atividades da sulfatase que retiram os radicais sulfatos ligados aos estrogênios naturais (estradiol e estrona) estão inibidas. Já as atividades da sulfotransferase que sulfatam os estrogênios naturais estão estimuladas, os inativando. A consequência é o desvio para as formas sulfatadas, que são as formas inativas dos estrogênios. Isto é um efeito direto da tibolona e de seus metabólitos ao nível de parênquima mamário. A tibolona inibe a proliferação de células epiteliais de tecido mamário normal e estimula a apoptose. Estudos clínicos têm mostrado que usuárias de tibolona apresentam menor sensibilidade e dor mamária e não mostram aumento da densidade mamográfica. A relação entre tibolona e risco de câncer de mama é inconclusiva e requer mais investigação

Tibolone is a tissue-specific hormone used for the treatment of climacteric symptoms and the prevention of osteoporosis in post-menopausal women. This work aimed at reviewing the effects of tibolone in the breast. Under action of tibolone, sulphatase activities, which take out sulfate binding to natural estrogens (estradiol and estrone) are inhibited. Sulphotransferase actions, which sulfate the natural estrogens are stimulated, inhibiting them. The consequence is the shift to inactive sulfated forms. This is a direct effect of tibolone and its metabolites in the mammary parenchyma level. Tibolone inhibits epithelial cells proliferation of normal breast tissue and stimulates apoptosis. Clinical researches have shown that tibolone users present low breast sensitivity and breast pain and they have not shown an increase in mammographic density. Tibolone and breast cancer risk are inconclusive and require further investigation

Tall stature and poor breast development after estrogen replacement in a hypergonadotrophic hypogonadic patient with a 45,X/46,X,der(X) karyotype with SHOX gene overdosage / Estatura alta e hipodesenvolvimento mamário após reposição estrogênica em paciente com hipogonadismo hipergonadotrófico e cariótipo 45,X/46,X, der(X) com superdosagem do gene SHOX

SHOX is exclusively expressed in the developing distal limb bones of human embryos and in the first and second pharyngeal arches. It works as a promoter for linear growth and as a repressor of growth plate fusion. It was reported, recently, that SHOX overdosage and gonadal estrogen deficiency have led to tall stature due to continued growth. We report, in the present study, a female patient with 45,X/46,X, psu idic(X)(pter→q21::q21→pter) karyotype, tall stature, and hypergonadotrophic hypogonadism without Turner stigmas. She did not present breast development even after long term therapy with high estrogen doses. Fluorescence in situ hybridization depicted the presence of three copies of SHOX gene. Microsatellite studies showed paternal origin of der(X). Further studies in similarly affected patients will clarify if the absence of breast development, despite previous high-dose estrogen treatment, is associated to triple copy of SHOX gene.

O gene SHOX é expresso, exclusivamente, no primeiro e no segundo arcos faríngeos, assim como nas extremidades dos ossos dos membros em embriões humanos. SHOX normalmente atua como um promotor para o crescimento linear e como um repressor do fechamento da placa de crescimento. Recentemente, foi descrito que o excesso da proteína SHOX associada à deficiência estrogênica gonadal leva à estatura alta devido ao contínuo crescimento. Neste estudo descrevemos uma paciente do sexo feminino com cariótipo 45,X/46,X,psu idic(X)(pter→q21::q21→pter), estatura alta, hipogonadismo hipergonadotrófico e sem estigmas de Turner. A paciente não apresentou desenvolvimento de mamas, mesmo depois do tratamento prolongado com altas doses de estrógenos. FISH evidenciou a presença de três cópias do SHOX. Estudo de microssatélites demonstrou a origem paterna do der(X). Estudos futuros em pacientes com semelhanças clínicas esclarecerão se a ausência de desenvolvimento de mamas, apesar do tratamento com altas doses de estrógenos, está associada à tripla cópia do SHOX.

Síndrome metabólica e câncer de mama em mulheres climatéricas / Metabolic syndrome and breast cancer in climateric women

O câncer de mama é um grave problema de saúde pública, que vem crescendo à medida que eleva sua incidência e a taxa de mortalidade. A natureza multifatorial dessa neoplasia é amplamente reconhecida, mas seus determinantes permanecem objeto de pesquisa e controvérsia; sendo assim, muito se tem pesquisado no sentido de identificar fatores que, controlados, atuem na redução da incidência e da recidiva dessa patologia. As pesquisas atuais têm sido dirigidas para a inter-relação entre alterações metabólicas e o câncer de mama, consideradas de caráter sistêmico ou mesmo no sítio mamário, admitindo o adipócito como sede de tais alterações. Entre os fatores metabólicos estão a obesidade, a hiperinsulinemia e , mais recentemente, a síndrome metabólica. Esta ultima consiste de uma conjunção de fatores que tem a resistência insulínica como substrato comum (hipertensão arterial, obesidade abdominal, hipoglicemia e dislipidemia), sendo outro grave problema que cresce na velocidade em que pioram os hábitos alimentares e aumenta o sedentarismo. No futuro, se pesquisas puderem confirmar essa associação, a manutenção do peso adequado poderá reduzir o riscode câncer de mama, e, por outro lado, a elucidação dessa associação poderá auxiliar no desenvolvimento de terapêutica medicamentosa capaz de reduzir o crescimento celular mediado pela hiperinsulinemia.

Breast cancer is a serious public health problem, which grows as its incidence and mortality rate raise. The multifactorial nature of this neoplasia is widely recognized, but its determinants remain controversy and object of researches. For this reason, much has been searched to identify factors, that controlled, can act upon incidence and recidive reduction of this pathology. Current researches have been directed to the interrelation between metabolic alterations and breast cancer, considered as systemic characteristics or located in mammary tissue, admitting adipocyte as the site of such alteration. Amongst the metabolic factors, there are: obesity, hyperinsulinemia and, more recently, metabolic syndrome. The last one consists on a conjunction of factors that has the insulin resistance as common substratum (arterial hypertension, abdominal obesity, hyperglycemia and dislipidemia), and is considered as another serious problem that grows with the same speed as alimentary habits get worse and sedentary increases. In the future, if the researches confirm this association, adequate weight maintenance will be able to reduce breast cancer risk, and, on the other hand, the briefing of this association will help the development of therapeutical drugs capable to reduce cellular growth mediated by hyperinsulinemia.

Expression of cyclooxygenase-2 and 12-lipoxygenase in human breast cancer and their relationship with HER-2/neu and hormonal receptors: impact on prognosis and therapy.

BACKGROUND: A number of studies have shown over-expression of cox-2 in breast cancer. Also it has been recorded that human breast cancer expresses high level of cox-2 and 12-lipoxygenase which may be beneficial in future therapy plan for those patients. AIMS: The present study aims to examine the level of transcripts of cox-2 and 12-lipoxygenase in Egyptian breast cancer patients and to compare between the expressions of both enzymes and TNM staging, hormone receptors status (including estrogen and progesterone) and HER2/neu expression. MATERIALS AND METHODS: Total cellular RNA was extracted from 64 frozen tissue samples of breast carcinoma and their corresponding normal adjacent tissues. Cox-2 and 12-lipooxygenase expressions were detected using RT-PCR. Hormonal receptors as well as HER2/neu were detected immuno-histochemically for each patient. RESULTS: About 47 and 62.5% of carcinoma samples showed over-expression of cox-2 and 12-lipooxygenase respectively as compared to their corresponding normal tissues. The results revealed that cox-2 significantly associated with TNM staging (P = 0.0047) and hormonal receptors status (P = 0.0201). The relationship between cox-2 and HER2/neu expression was close to a significant value (P = 0.0747). 12-lipooxygenase showed only significant association with TNM staging (P = 0.0076). Neither hormonal receptors nor HER2/neu showed significant association with this enzyme. CONCLUSION: Elevated levels of cox-2 and 12-lipoxygenase expression were detected in human breast cancer. Also, the results revealed that cox-2 and 12-lipooxygenase mRNA expressions are associated with TNM staging in human breast cancer. Furthermore, there is an inverse association between cox-2 expression and hormonal receptor status. This observation may drive us to the possible role of those two enzymes in determining the plan of therapy of breast cancer patients.

Genetic polymorphism of CYP17 and breast cancer risk in Korean women

CYP17 gene is involved in steroidogenesis and steroid metabolism. Epidemiologic results on the association between the CYP17 polymorphism and breast cancer risk have been inconsistent. We examined the association between the MspAI polymorphism at +27 relative to the start of transcription in the 5'-untranslated region of CYP17 gene and breast cancer risk in Korean women. Four hundred and sixty-two incident cases and 337 controls were recruited from three teaching hospitals in Seoul during 1994-2001. Polymorphism of the CYP17 gene was determined by a single base extension assay. Demographic and lifestyle characteristics were identified using structured questionnaire. Age-adjusted (aOR) and multivariate odds ratios (mOR) and 95% confidence intervals (CI) were estimated by unconditional logistic regression. The proportions of A1/A1, A1/A2 and A2/A2 genotypes among controls were 20.8%, 45.1% and 34.1%, respectively. Compared to the A1/A1 genotype, A1/A2 or A2/A2 genotype was not statistically significantly associated with overall breast cancer risk (i.e., mOR=1.01, 95% CI=0.69-1.47 and mOR=0.76, 95% CI=0.51-1.14, respectively). However, a significant association between CYP17 A2/A2 genotype and breast cancer was observed among women aged 50 years or less (mOR=0.58, 95% CI=0.34-0.99, P=0.04) and leaner women (body mass index < 22 kg/m2) (mOR=0.48, 95% CI=0.23-0.97, P=0.04). Our results suggest that genetic polymorphism in 5'-untranslated region of CYP17 might play a role in breast cancer development in Korean women among younger women aged less than 50 or leaner women with body mass index less than 22 kg/m2.

Overexpression of c-erbB-2 oncoprotein and associated pathobiological factors in invasive primary breast cancer.

Breast cancer tissues from 271 cases were analyzed immunologically for c-erbB-2 oncoprotein (HER-2/neu), epidermal growth factor receptor (EGF-R) and estrogen receptor (ER). Overexpression of both c-erbB-2 oncoprotein and EGF-R showed an inverse association with ER and a direct association with metastatic involvement of lymph node and high histological grade. The frequency of c-erbB-2 and EGF-R overexpression was significantly higher among postmenopausal cases in comparison with premenopausal cases. Further, only in postmenopausal patients, c-erbB-2 oncoprotein (chi2 = 6.4, P < 0.05) and EGF-R (chi2 = 6.4, p < 0.05) as well as their concomitant expression (chi2 = 11.5, p < 0.01) revealed a statistically significant association with ER.

Immunohistochemical localization of transferrin in human breast cancer tissue.

The present study was planned to detect the iron binding protein, transferrin (TR) in paraffin sections of the human breast tumors. The distribution of transferrin has been studied in 153 cases (63 benign lesions and 90 malignant tumors). The extent of staining reaction was determined by semiquantitative grading (weak, moderate and consistent). Positivity rate for transferrin was higher (92.2%) in malignant tumors as compared to benign breast lesions (28.5%) with significant p value (p = 0.0001) for both the groups. The intensity was variable in both the groups, being more intense in the malignant tumors. Tumors with higher grade of malignancy presented consistent positive staining along with the lymph nodes involved. The extent of immunoreactivity revealed a significant positive correlation with axillary lymph node status. However, no significant correlation was found with the age of the patients. Thus the study of transferrin in breast tumors besides being of prognostic significance helps in the further management of malignant lesions of the breast.

Expression of cyclins in ductal hyperplasia, atypical ductal hyperplasia and ductal carcinoma in situ of the breast

Cyclin/cdc complexes are known to function in cell-cycle regulation. Cyclin D1/cdk4 and -6 complexes, which functions as a G1-S checkpoint and cyclin B1/cdc2 complexes, a G2-M checkpoint are essential for DNA synthesis and mitosis, respectively. Thus, dysregulated overexpression of cyclins appears to be involved in uncontrollable cell proliferation and early tumor development. We investigated the expression and proliferative index of cyclin D1 (PIcyclin D1), cyclin B1 (PIcyclin B1) and Ki-67 (PIKi-67) using immunohistochemical staining on 15 cases of ductal hyperplasia (DH), 26 cases of atypical ductal hyperplasia (ADH) and 43 cases of ductal carcinoma in situ (DCIS) of the breast in order to evaluate whether these cyclins are associated with abnormal cell proliferation and play a role in tumor development from ADH to carcinoma. Furthermore, we investigated whether the expression and proliferative index of the cyclins and Ki-67 are correlated with the histologic grade according to the Van Nuys classification and with the histologic subtype according to traditional classification. Finally, we estimated the correlation coefficient among PIcyclin D1, PIcyclin B1, PIKi-67 and estrogen receptor in ADH and DCIS. The expression of cyclin D1 was detected in 39.5% of DCIS and 7.7% of ADH cases. In the DH cases, expression of cyclin D1 was not found. Expression of cyclin B1 was also detected in 69.7% of DCIS, 50.0% of ADH and 93.3% of the DH cases. The PIcyclin D1 was significantly different among these three groups. Moreover, the PIcyclin D1 and PIKi-67 were differed significantly between the low grade DCIS and ADH cases. However, PIcyclin B1 only appeared to be significantly different between the total DCIS and ADH. Results of the correlation coefficient among PIcyclin D1, PIcyclin B1 and PIKi-67 were positively correlated with each other. No significant correlation was found between the expression of ER and cyclin D1 in ADH and DCIS. In summary, our results support the hypothesis that a cyclin D1 and cyclin B1 protein aberration, along with Ki-67, may act as a relatively early event in the tumor development from ADH to carcinoma.

Alanine aminotransferase activity in mammary tissue, muscle and liver of dam rat during lactation and weaning

Transamination reaction is the first step in the catabolism of most of the L-amino acids. Alanine is an important molecule in the inter-organ nitrogen transport, conveying them from muscle to the liver Amino groups from this amino acid are generally first transferred to alfa-ketoglutarate in the cytosol of liver cells to form glutamate and leaving behing the corresponding alfa-keto acid analog. Measurements of the alanine aminotransferase (EC2.6.1.2.) activity were compared in liver, mammary gland an skeletal muscle in virgin, lactating and weaning dam rats. In this study liver was the principal tissue involved in alanine transamination, while muscle showed a reduction in the enzyme activity during lactation. Results indicate an increase in alanine aminotransferase activity in the mammary gland during lactation and weaning when compared with virgin rats. This suggest that mammary gland during lactation is an importan extra-hepatic tissue involved in the metabolism of alanine and probably shunted into hte pathways for amino group metabolism in terms of nitrogen economy

Bauhinia purpurea and Pisum sativum lectin binding in human breast.

The binding of biotinylated bauhinia purpurea (BPA) and pisum Sativum (PSA) lectins to paraffin-embedded tissue sections from 10 normal breast and 55 breast carcinoma has been investigated by applying avidin biotin peroxidase method (ABC). BPA showed very low affinity for normal breast epithelium and the binding was confined to the luminal surface. Eighty-seven percent carcinoma bound BPA, and the staining patterns varied depending on the histologic grade of tumors: luminal surface binding in grade 1 carcinomas; diffuse, granular cytoplasmic with para- or perinuclear deposits and staining along the plasma membrane in grade 2 and grade 3 carcinomas. PSA bound consistently to the luminal surface of all ducts and acinar cells of normal breast tissue. PSA was reactive with all carcinoma but the staining profiles were similar regardless of the tumor differentiation. It is concluded that the lectins used in this study have limited usefulness in routine diagnosis.

Detection of estrogen receptor in formalin-fixed and paraffin-embedded breast carcinoma: correlation with histological patterns

The authors studied the expression of estrogen receptor (ER) in tissues of breast carcinomas which were previously fixed in formalin and paraffin-embedded. The ER expression was correlated with several histological findings, namely grade of differentiation, tumor necrosis, desmoplasia, lymphocytic infiltration and elastosis. The ER was detected in tissues using the avidin-biotin immunoperoxidase technique associated with the H222 monoclonal antibody from Abbott. All 39 biopsies were of infiltrating ductal carcinoma of breast and 16 of them expressed ER. The statistical analysis showed that the expression of ER was correlated with histological findings of good prognosis as well differentiated carcinomas, no tumor necrosis, absence or mild lymphocytic infiltration around the tumor cells and severe elastosis

Receptores hormonais / Hormonal receptors

Os receptores de estrogênio, progesterona e androgênios têm sido pesquisados e apresentam aplicaçäo prática importante com relaçäo ao tratamento de doenças hormônio-dependentes. Este artigo apresenta uma revisÒo de literatura sobre o modo de açäo, métodos de análise e valor prognóstico desses receptores hormonais em Ginecologia.

Aspectos radiológicos das lesöes mamárias com secreçäo papilar: importância da ductografia / Radiological aspects of the mammary injuries with papillary secretion: ductography importance

Por ser um exame fundamental para o estabelecimento do plano terapêutico em pacientes com descarga papilar espontânea, a ductografia, tal como é realizada no INCa, é descrita em seus procedimentos e análises. A partir da reuniäo de treze casos registrados naquela instituiçäo, os elementos definidores do correto diagnóstico, assim como os procedimentos complementares utilizados, säo demonstrados de modo simples e objetivo