O6-Methyguanine-DNA Methyl Transferase (MGMT) Promoter Methylation in Serum DNA of Iranian Patients with Colorectal Cancer

Introduction: Colorectal cancer (CRC) is a leading cause of cancer deaths worldwide but current molecular targetedtherapy is not providing major success in CRC treatment, so early detection by non-invasive methods continues tobe vital. Aberrant methylation of CpG islands in promoter regions is associated with inactivation of various tumorsuppressor genes. O6-methyguanine-DNA methyltransferase (MGMT) is a DNA repair enzyme that removes mutagenicand cytotoxic adducts from O6-guanine in DNA. Aberrant hypermethylation of the MGMT promoter has beenassociated with lack of mRNA expression, with concomitant loss of protein content and enzyme activity. AIM: Ouraim was to determine whether MGMT promoter methylation might be detectable in circulating free DNA in the serumof CRC patients and normal individuals using a methylation specific (MSP) polymerase chain reaction (PCR) method.Methods: A total of 70 subjects were enrolled in the study. Of these, 30 patients who were diagnosed previously asuntreated colon adenocarcinoma by a gastroenterologist and the other 40 were nearly age-matched individuals who hada normal colonoscopic evaluation (except for hemorrhoids or fissures) and normal pathologic reports. After bisulphitemodification of DNA, serum samples were examined for MGMT promoter methylation using MSP. Results: Ninetypercent of CRC patients had MGMT promoter hypermethylation as compared to no methylation in normal subjects’serum. Most of the cancers were stage П and moderately differentiated adenocarcinomas; nearly 60% were found inthe left colon. No statistically significant correlation was found between the promoter methylation status and genderand age. Discussion and Conclusions: MGMT hypermethylation can be detected in free circulating DNA in serum ofCRC patients and can be used “as a clinical biomarker” for early diagnosis and prognostic assessment of the disease.Our data confirm previous studies indicating utility for free circulating DNA as a serum biomarker for early detection,diagnosis and monitoring of CRC patients.     

Expression of Estrogen Receptor Alpha and Beta in Breast Cancers of Pre- and Post-menopausal Women

Expression of estrogen receptors (ER) is clinically relevant in designing therapeutic strategies. The relative importance of the two types of estrogen receptors (ER-alpha and ER-beta) in human breast cancers in pre- and post-menopausal women has not been properly defined. To determine the possible association between the expression of estrogen receptor and serum estradiol levels in pre- and post-menopausal women with breast cancer. 44 patients with invasive ductal carcinoma of the breast were studied and a breast tissue biopsy was taken. ER-alpha and ER-beta were detected by immunocytochemistry. Serum levels of estradiol and estrone were measured by radioimmunoassay and FSH was measured using IRMA. We studied 21 pre- and 23 post-menopausal women with breast carcinoma. Examining the number of cases with tumors positive for ER, we found no differences in the frequency of ER-alpha between pre- and post-menopausal women, but ER-beta decreased marginally after menopause (p < 0.051). In cases with tumors positive for ER, the proportion of cells positive for ER-alpha was similar post-menopausally (53.95%) and pre-menopausally (57.21%), but for ER-beta the number of positive cells decreased significantly after menopause (p < 0.051). In pre-menopausal women there was a correlation between serum estradiol levels and ER-beta; in post-menopausal women there was a correlation between serum FSH levels and ER-alpha. These results indicate that estradiol levels in women with mammary carcinoma are related to ER-beta expression in the breast tumor tissue.     

原发性乳腺癌组织中胞浆,胞核受体测定分析

本文使用0.6M KC1法对61例原发性乳腺癌的组织测定核受体(ERn)。结果,25例ERn~(( ))占40.9％;36例ERn~((-))占59.1％,同时与胞浆受体(ERc)进行了比较。ERc、ERn皆为阳性占34.4％;皆为阴性占29.5％。另外,我们发现ERc、ERn之间的关系趋向于胞浆受体阳性患者,其核亦可为阳性,而胞浆受体阴性者,核受体阳性却为少见现象。     

Estrogen Receptor Alpha Gene Polymorphisms and Breast Cancer Risk: a Case-control Study with Meta-analysis Combined

Molecular epidemiological studies have shown that gene polymorphisms of estrogen receptor alpha gene(ESR-α) are associated with breast cancer risk. However, previous results from many molecular studies havebeen inconsistent. In this study, we examined two polymorphisms (PvuII and XbaI RFLPs) of the ESR-αgene in 542 breast cancer cases and 1,016 controls from China. Associations between the polymorphisms andbreast cancer risk were calculated with an unconditional logistic regression model. Linkage disequilibrium andhaplotypes were analyzed with the SHEsis software. In addition, we also performed a systematic meta-analysisof 24 published studies evaluating the association. No significant associations were found between the PvuIIpolymorphism and breast cancer risk. However, a significantly decreased risk of breast cancer was observedamong carriers of the XbaI ‘G’ allele (age-adjusted OR = 0.80; 95% CI = 0.66- 0.97) compared with carriers ofthe ‘A’ allele. Haplotype analysis showed significantly decreased cancer risk for carriers of the ‘CG’ haplotype(OR = 0.79; 95% CI = 0.66- 0.96). In the systematic meta-analysis, the XbaI ‘G’ allele was associated with anoverall significantly decreased risk of breast cancer (OR = 0.90, 95% CI = 0.82- 1.00). In addition, the PvuII‘C’ allele showed a 0.96- fold decreased disease risk (95% CI = 0.92- 0.99). In subgroup analysis, an associationbetween the PvuII ‘C’ and XbaI ‘G’ alleles and breast cancer risk was significant in Asians (‘C’ vs. ‘T’: OR =0.93, 95% CI = 0.85- 1.00; ‘G’ vs. ‘A’: OR = 0.82, 95% CI = 0.68- 0.98), but not in Euro-Americans. Thus, ourresults provide evidence that ESR-α polymorphisms are associated with susceptibility to breast cancer. Theseassociations may largely depend on population characteristics and geographic location.     

雌激素受体GPER在乳腺癌中的研究现状

G蛋白偶联雌激素受体（G protein-coupled estrogen receptor, GPER, 也称GPR30）在多种类型细胞中均可介导雌激素信号,其在乳腺癌、子宫内膜癌等激素敏感性肿瘤细胞中的作用尤为明显。雌激素及抗雌激素药物通过激活GPER促进下游信号通路的活化及靶基因的表达进而参与乳腺癌细胞的增殖、迁移、侵袭及他莫昔芬耐药等恶性生物学行为。目前发现表皮生长因子受体（epidermal growth factor receptor, EGFR）的转活是GPER引发下游生物学效应的关键靶点。此外,GPER还被认为是预测三阴乳腺癌患者预后的潜在生物学标志物之一,对GPER的深入研究可能为乳腺癌患者的综合性评估和临床治疗打开更广阔的前景。     

术前化疗对晚期乳腺癌雌激素受体含量的影响

目的：探讨术前化疗对晚期乳腺癌雌激素受体含量的影响。方法：42例晚期乳腺患者分为两组。A组（31例）：术前接受Cooper‘s方案化疗周。B组（11例）；术前未进行化疗。术后对两组乳腺癌组织用MCG法测定雌激素受体含量，并比较两组之间有无显著差异。结果：A组雌激素受体平均含量为57.00fmol/mg蛋白质，B组雌激素受体平均含量为12.58fmol/mg蛋白质，A组与B组差异有显著意义（P＜0.005)。结论：经术前化疗后雌激素受体含量较高，因此有必要在化疗前测定乳腺癌患者的雌激素受体含量，以便使所测得值准确地反映乳腺癌患者癌组织的真实受体状态，合理选择适当的治疗措施。     

Association of miR-1266 with Recurrence/Metastasis Potential in Estrogen Receptor Positive Breast Cancer Patients

The Homeobox B13 (HOXB13):Interleukin 17 Receptor B (IL17BR) index of estrogen receptor (ER)-positivebreast cancer (ER (+) BC) patients may be a potential biomarker of recurrence/ metastasis. However, effects ofmicroRNA (miRNA) binding to the 3’ untranslated region (3´UTR) of HOXB13 and IL17BR and its function onrecurrence/metastasis in ER (+) BC remains elusive. The aims of this study were to determine the expressionof miRNAs that bind to 3´UTR of HOXB13 and IL17BR in ER (+) BC patients and asess the effects of thesemiRNAs on recurrence/metastasis. The expression profiles of HOXB13 and IL17BR were evaluated using RTPCRin tumors and normal tissue samples from 40 ER (+) BC patients. The expression level of 4 miRNAs, whichwere predicted to bind the 3´UTR of HOXB13 and IL17BR using TargetScan, microRNA.org and miRDB onlinedatabases, were further evaluated with RT-PCR. Our findings demonstrated that high miR-1266 levels might besignificant prognostic factor for recurrence/metastasis occurrence (3.05 fold p=0.004) and tamoxifen response(3.90 fold; p=0.2514) in ER (+) BC cases. Although we suggest that modulation of miR-1266 expression may bean important mechanism underlying the chemoresistance of ER (+) BC, advanced studies and validation arerequired.     

Clinico-Pathologic Subtypes of Breast Cancer Primary Tumors Are Related to Prognosis after Recurrence

Background: Pathological factors, based mainly on immunohistochemistry (IHC) and histological differentiation, are mostly used to differentiate breast cancer (BC) subtypes. Our present aim was to describe the characteristics and survival of a relapsing BC patient cohort based on clinico-pathologic subtypes determined for the primary tumors. Methods: We used a clinico- pathological definition of BC subtypes based on histological grade (HG), estrogen receptor (ER), progesterone receptor (PgR),and epidermal growth factor receptor type 2 (HER2) expression assessed by IHC. We determined variables associated with loco-regional recurrence (LRR), second primaries (SP), systemic recurrence (SR) and post-recurrence survival (PRS). Results: Out of 1,702 patients, 240 (14%) had an event defined as recurrence. Those with recurrent disease were significantly younger than those without,and were initially diagnosed at more advanced stages, with larger tumors, greater lymph nodal involvement and higher HG. With a median follow up of 61 months (1-250), 4.6% of patients without recurrence and 56.6% of patients with an event defined as recurrence had died. The median PRS for the LRR group was 77 months; 75 months for those who developed a SP and 22 months for patients with an SR (p <0.0001). In SR cases, the median PRS was shorter for ER- tumors than for ER+ tumors (15 vs. 26 months, respectively; p = 0.0019, HR 0.44; CI: 0.25-0.44). Conclusions: Subtype, defined through classic histopathologic parameters determined for primary tumors, was found to eb related to type of recurrence and also to prognosis after relapse.     

Factors Influencing Breast Cancer Screening Behavior among Iranian Women

Background: Early detection of breast cancer is of great importance to improve women’s health and to decreasethe cost related to cancer death. Therefore, recognition of variables related to breast cancer screening behaviorsis necessary. Objectives of this study were to identify the rates of breast self-examination (BSE) performance andmammography use in Iranian women, and to characterize the demographic and cognitive factors associated withtheir breast cancer screening behavior. Method: Data were collected from a convenience sample of 388 females,using an adapted version of Champion’s revised Health Belief Model Scale. Results: The results showed that7.5% of the participants performed BSE on a regular monthly basis, and among the women aged 40 and older,14.3% reported having had at least one mammography in their lifetime. Perceived self-efficacy and perceivedbarriers to BSE were significant predictors for BSE performance. For having mammography, health motivationwas the main predictor. Conclusion: Eliminating barriers and increasing perceived self-efficacy with an emphasisto make the women acquainted with BSE performance; as well as increasing health motivation of women andpersuading of physicians for clinical breast examination (CBE) performance with low cost and free access tom ammography, are important to promote BSE and mammography.     

Association of Estrogen Receptor Alpha and Interleukin 6 Polymorphisms with Lymphovascular Invasion,Extranodal Extension,and Lower Disease-Free Survival in Thai Breast Cancer Patients

Breast cancer is the most frequent type of cancer diagnosed among women worldwide and also in Thailand. Estrogen and estrogen receptors exert important roles in its genesis and progression. Several cytokines have been reported to be involved in the microenvironment that promotes distant metastasis via modulation of immune and inflammatory responses to tumor cells. Estrogen receptor genetic polymorphisms and several cytokines have been reported to be associated with breast cancer susceptibility and aggressiveness. To investigate roles of genetic polymorphisms in estrogen receptor alpha (ESR1) and interleukin 6 (IL6), breast cancer patients and control subjects were recruited from the Division of Head, Neck and Breast Surgery (Siriraj Hospital, Bangkok, Thailand). Polymorphisms in ESR1 (rs3798577) and IL6 (rs1800795 and rs1800797) were evaluated by real-time PCR in 391 breast cancer patients and 79 healthy controls. Associations between genetic polymorphisms and clinicopathological data were determined. There was no association between genetic polymorphisms and breast cancer susceptibility. However the ESR1 rs3798577 CT genotype was associated with presence of lymphovascular invasion (OR=2.07, 95%CI 1.20-3.56, p=0.009) when compared to the TT genotype. IL6 rs1800795 CC genotype was associated with presence of extranodal extension (OR= 2.30, 95%CI 1.23-4.31, p=0.009) when compared to the GG genotype. Survival analysis showed that IL6 rs1800797 AG or AA genotypes were associated with lower disease-free survival. These findings indicate that polymorphisms in ESR1 and IL6 contribute to aggressiveness of breast cancer and may be used to identify high risk patients.     

Dietary Habits Contributing to Breast Cancer Risk Among Iranian Women

Background: The aim of this study was to investigate demographic features, dietary habits, and some possiblerisk factors for being susceptible to breast cancer in Iranian women. Materials and Methods: A study of dietaryhabits and breast cancer was conducted among 53 Iranian women with histological confirmed disease and 40matched controls. A dietary habits questionnaire was used to evaluate the pattern of selected food intakes. Therisk of cancer was analyzed after adjustment for confounding factors. Age, weight, body mass index (BMI), waistcircumference, educational status, parity, lactation, marital status, menopause, history of estrogen therapy, andfamily history of breast disease or cancer were assessed among participants. Special attention was given to therelationship between consumption of high fat meat, milk, yogurt and cheese as well use of frying oils for fryingfoods, use of olive/liquid oils for cooking, removing fat from meat and poultry, removing chicken skin and not useof mayonnaise as salad dressing and the risk of breast cancer. Moreover, salad, vegetable and fruit consumption,and eating outdoors owere investigated. Results: Our results revealed significant lower education and higher BMIand waist circumference levels in patients with breast cancer. There was significantly increased breast cancerrisk in overweight women in comparison with normal weight (OR=2.91, 95%CI 1.24 to 6.82). High intake of fatdairy products including milk and cheese was found to be a statistically significant factor for increasing breastcancer risk in models adjusting for age, BMI and education. Use of olive/liquid oils for cooking and avoidanceof mayonnaise as salad dressing are related to lower risk of breast cancer. The frequency of vegetable and fruitconsumption was significantly lower in patients with breast cancer compared to healthy women. Conclusions:Dietary habits might be risk factors for breast cancer among Iranian women. Adoption of a prudent diet couldbe an appropriate strategy for preventing breast cancer.     

乳腺癌组织中孕激素受体与雌激素受体DNA结合功能关系的研究

目的：观察３６例（Ｅ２＋）乳腺癌患者组织ＰＲ和ＥＲ ＤＮＡ结合功能的关系,进而探讨（Ｅ２＋／ＰＲ＋）表型内分泌治疗无反应的分子机制。方法：用激素结合法和迁移率改变法。结果：１）用含有ＥＲ的ＭＣＦ－７细胞株作为阳性对照,２２℃Ｍｇ＾２＋存在条件下,迁移率改变法中ＥＲ－ＥＲＥ复合物的形成是激素依赖性的,证实了ＥＲ－ＥＲＥ复合物的特性。（２）激素结合法和迁移率改变法检测结果显示,３６例（Ｅ２＋）乳腺癌中     

Promoter Methylation of BRCA1, DAPK1 and RASSF1A is Associated with Increased Mortality among Indian Women with Breast Cancer

Background: Promoter methylation has been observed for several genes in association with cancer development and progression. Hypermethylation mediated-silencing of tumor suppressor genes (TSGs) may contribute to breast cancer pathogenesis. The present study was conducted to investigate the promoter methylation status of BRCA1, DAPK1 and RASSF1A genes in Indian women with breast cancer. Materials and Methods: Promoter methylation was evaluated in DNA extracted from mononuclear cells (MNCs) in peripheral blood samples of 60 histopathologically confirmed newly diagnosed, untreated cases of breast cancer as well as 60 age and sex matched healthy controls using MS-PCR. Association of promoter methylation with breast cancer-specific mortality was analyzed with Cox proportional hazards models. Kaplan-Meier survival analysis was performed for overall survival of the breast cancer patients. Results: We observed a significant increase of BRCA1, DAPK1 and RASSF1A promoter methylation levels by 51.7% (P <0.001), 55.0% (P <0.001) and 46.6% (P <0.001), respectively, when compared to healthy controls. A strong correlation was noted between hypermethylation of the tumor suppressor genes BRCA1 (P= 0.009), DAPK1 (P= 0.008) and RASSF1A (P= 0.02)) with early and advanced stages of breast cancer patients. We also found that breast cancer-specific mortality was significantly associated with promoter methylation of BRCA1 [HR and 95% CI: 3.25 (1.448-7.317)] and DAPK1 [HR and 95% CI: 2.32 (1.05-5.11)], whereas limited significant link was evident with RASSF1A [HR and 95% CI: 1.54 (0.697-3.413]. Conclusion: Our results suggest that promoter methylation of BRCA1, DAPK1 and RASSF1A genes may be associated with disease progression and poor overall survival of Indian women with breast cancer.     

Estrogen Receptor Alpha Gene Expression in Breast Cancer Tissues from the Iranian Population - a Pilot Study

Estrogen receptor alpha (ERα) is one of the major sub-types of estrogen receptors. ERα plays an important rolein cellular proliferation and differentiation, chiefly in mammary tissues. In the present study we aimed to quantifyof ERα mRNA and protein expression in breast tissues from the Iranian population using a real-time PCR assay.Twenty nine breast tissues including 19 adenocarcinomas and 10 normal controls were recruited from the Iranianpopulation. mRNA extraction and cDNA synthesis were performed from these tissues using commercial kits.ERα mRNA and protein expression was quantified using real-time PCR and immunohistochemistry respectively.The results showed high expression of ERα mRNA (68%) and protein (53%) in the majority of breast cancertissues compared to normal breast tissues (p= 0.035). Also, high ERα mRNA was associated with tumour size ofbreast carcinomas. In this study, we first reported the expression of ERα in Iranian patients with breast cancersand demonstrated prevalence of the expression to be similar to breast cancers noted in other populations.     

Survival Outcomes in Breast Cancer Patients With Low Estrogen/Progesterone Receptor Expression

IntroductionThe prognostic value of low estrogen and progesterone receptors expression (ER/PgR 1%-10%) in early breast cancer patients is still unclear.Patients and MethodsWe retrospectively analyzed 1424 consecutive patients with HER2/neu-negative and low endocrine receptors expression early breast cancer, submitted to surgery at the European Institute of Oncology between January 1995 and December 2009. Patients were classified according to the percentage of ER/PgR expression using immunohistochemistry. Group 1 with ER/PgR < 1%, and group 2 with ER/PgR 1% to 10%.ResultsGroup 1 (ER/PgR < 1%) included 1300 patients, and group 2 (ER/PgR 1%-10%) 124 patients. Median follow-up time was 74 months (range, 3-192 months). The 5-year disease-free survival (DFS) rate was 74% (95% confidence interval [CI], 72%-77%) for group 1, and 79% (95% CI, 70%-86%) for group 2 (P = .16). The 5-year overall survival (OS) rate was 86% (95% CI, 84%-88%) in group 1 and 90% (95% CI, 83%-95%) in group 2 (P = .13). In patients without lymph node involvement, the 5-year OS rate was 92% (95% CI, 89.5%-93.6%) for group 1 and 98% (95% CI, 90.2%-99.8%) for group 2 (P = .061). One hundred ten patients received endocrine therapy with no significant effect on DFS (P = .36) and OS (P = .30).ConclusionThe ER/PgR 1%-10% group had a slight, but not statistically significant, better prognosis than the ER/PgR <1% group. Further studies are needed to identify the appropriate clinical approach in this subset of patients with low ER/PgR expression (ER/PgR 1%-10%), HER2-negative early breast cancer.     

MicroRNA在雌激素受体阳性乳腺癌中的研究进展

MicroRNA(miRNA)能调节细胞生长、增殖、分化和凋亡等活动,与各种恶性肿瘤的发生发展息息相关,乳腺癌亦不例外.在乳腺癌中,雌激素受体(ER)阳性者占有相当大的比例,约75％,因此雌二醇(E2)、ER与miRNA的相互联系以及相互调节机制需予以重视,进而有助于探索miRNA对ER阳性乳腺癌的临床应用价值.全文就近年来的相关研究成果加以概述和总结,旨在为转化医学提供信息,将相关研究成果应用于临床.     

Estrogen Receptor and Progesterone Receptor Status in Breast Cancer in Relation to Age,Histological Grade,Size of Lesion and Lymph Node Involvement

Introduction: Breast cancer is the most common malignancy of women in Kashmir. This study was conductedwith the objective of assessing hormone receptor positivity and its correlation with age at diagnosis, tumor size,histological grade and lymph node metastasis. Materials and Methods: 132 newly diagnosed cases of invasivebreast cancer diagnosed at the Department of Pathology, SKIMS, Srinagar, J&K, were included after excludingbiopsies, in-situ lesions and recurrence cases. Results: Mean age of the patients was 48.2 years, 59.1% being≤50 years of age. Mean duration of symptoms was 6.32 months. Most lesions (65.1%) were 2-5 cm and 16.7%were ≥5.0 cm in greatest dimension. The predominant (80.3%) morphology was IDC-NOS. The majority of thecases presented as grade II (52.1%) lesions and lymph node involvement was present in 65.2%. ER and PRwere positive in 66.3% and 63.4% cases, respectively, increasing with rising age. High grade lesions and largersize tumors were more likely to be ER and PR negative. No correlation was found between ER/PR status andlymph node metastasis. Conclusions: ER and PR expression in breast cancers in the current study was found tobe higher than studies done in India/Asia but lower than studies conducted in the West, even on Indian/Asianimmigrants. Markedly lower receptor expression in Indian/Asian studies is likely due to preanalytic variables,thresholds for positivity, and interpretation criteria. American Society of Clinical Oncology/College of AmericanPathologists Guideline Recommendations for Immunohistochemical Testing of Estrogen and ProgesteroneReceptors in Breast Cancer are strongly advocated for standardization of receptor evaluation and for clinicalmanagement of breast cancer patients to provide best therapeutic options.     

Ultrasonographic Features of Triple-Negative Breast Cancer: a Comparison with Other Breast Cancer Subtypes

Background: Triple-negative breast cancer (TNBC) is known to be associated with aggressive biologic featuresand a poor clinical outcome. Therefore, early detection of TNBC without missed diagnosis is a requirementto improve prognosis. Preoperative ultrasound features of TNBC may potentially assist in early diagnosis ascharacteristics of disease. Purpose: To retrospectively evaluate the sonographic features of TNBC compared toER (+) cancers which include HER(-) and HER2 (+), and HER2 (+) cancers which are ER (-). Materials andMethods: From June 2012 through June 2014, sonographic features of 321 surgically confirmed ER (+) cancers(n=214), HER2 (+) cancers (n=66), and TNBC (n=41) were retrospectively reviewed by two ultrasound specialistsin consensus. The preoperative ultrasound and clinicopathological features were compared between the threesubtypes. In addition, all cases were analyzed using morphologic criteria of the ACR BI-RADS lexicon. Results:Ultrasonographically, TNBC presented as microlobulated nodules without microcalcification (p=0.034). A lowerincidence of ductal carcinoma in situ (p<0.001), invasive tumor size that is>2 cm (p=0.011) and BI-RADS category4 (p<0.001) were significantly associated with TNBC. With regard to morphologic features of 41 TNBC cases,ultrasonographically were most likely to be masses with irregular (70.7%) microlobulated shape (48.8%), becircumscribed (17.1%) or have indistinct margins (17.1%) and parallel orientation (68.9%). Especially TNBCmicrolobulated mass margins were more more frequent than with ER (+) (2.0%) and HER2 (+) (4.8%) cancers.Conclusions: TNBC have specific characteristic in sonograms. Ultrasonography may be useful to avoid misseddiagnosis and false-negative cases of TNBC.