فهرست مطالب

Reports of Biochemistry & Molecular Biology - Volume:8 Issue:1, 2019
  • Volume:8 Issue:1, 2019
  • تاریخ انتشار: 1398/02/11
  • تعداد عناوین: 15
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  • Mahdieh Fatemi Nayeri, Ali Talaei*, Jalil Tavakkol Afshari, Amin Reza Nikpoor, Andisheh Talaei, Rashin Ganjali Pages 1-8
    Background
    The pathophysiology of bipolar 1 disorder (B1D), a major psychiatric disorder with inflammatory origins and structural changes in the brain, is of great interest to researchers. Pro-inflammatory biomarkers and specific gene expression play pivotal roles in B1D development, and IFN-γ has emerged as an important inflammatory marker. The aim of this research was to determine whether the INF-γ +874 T/A polymorphism is associated with B1D susceptibility in an ethnic Iranian population.
    Methods
    The IFN-γ +874 T/A (rs2430561) gene polymorphism was studied in 106 B1D patients and 109 control subjects using sequence specific primers (SSPs) and amplification refractory mutation system-polymerase chain reaction (ARMS-PCR).
    Results
    Significant statistical differences in IFN-γ +874 T/A polymorphism genotype distribution were found between the patients and control subjects (P = 0.0006). Decreased risk of B1D was detected in the codominant model (T/T vs T/A and A/A, OR = 0.19, 95% CI = 0.07-0.49 for T/A, OR = 0.38, 95% CI = 0.12-1.24 for A/A, P value=0.0006), and in the dominant model (T/T vs T/A-A/A, OR = 0.21, 95% CI = 0.08-0.54, P = 0.0005). However, no significant difference in the IFN-γ polymorphism allele distribution was found between the two groups (P = 0.25).
    Conclusions
    The IFN-γ +874 T/A polymorphism may have a significant role in BID development.
    Keywords: Bipolar 1 Disorder, Gene Polymorphism, Interferon gamma
  • Mohammad Mahdi Hayatbakhsh, Arezoo Gowhari Shabgah, Saeed Pishgouyi, Jalil Tavakiol Afshari, Hadi Zeidabadi, Mojgan Mohammadi* Pages 9-14
    Background
    Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder characterized by altered bowel habits and abdominal pain in the absence of a recognizable structural anomaly. The pathogenesis of IBS has been associated with inflammation and the expression of pro-inflammatory chemokines, such as CCL2 and CCL16. Our study aimed to investigate the relationship between the serum levels of CCL2 and CCL16 and IBS. Additionally, we examined how serum levels of these chemokines relate to IBS subtypes.
    Methods
    Patients with IBS diagnosed according to the Rome III criteria participated in this study (n= 96). Healthy individuals with no history of allergic, autoimmune, chronic or active gastrointestinal infectious diseases were used as controls (n= 44). The serum levels of CCL2 and CCL16 was measured via enzyme-linked immunosorbent assay (ELISA).
    Results
    A significant decrease in the serum levels of CCL16 and CCL2 was observed in the patients with IBS. Additionally, the serum levels of CCL16 in IBS patients with diarrhea (D-IBS) was significantly higher than those with the mixed IBS (M-IBS) subtype.
    Conclusions
    The significant increase in the serum levels of CCL-16 in patients with D-IBS compared to patients with M-IBS suggests that CCL-16 may be used as an immunological biomarker to differentiate between these two subtypes.
    Keywords: CCL-2, CCL-16, C-IBS, D-IBS, MCP-1, M-IBS, IBS
  • Amin Mahmoudi, Keihan Ghatreh Samani*, Seyed Asadollah Amini, Esfandiar Heidarian Pages 15-20
    Background
    Pioglitazone increases insulin sensitivity and improves glycemic control in type 2 diabetics. In this study, we evaluated the effects of pioglitazone on the uncoupling protein 1 (UCP1) expression in mouse brown adipose tissue (BAT), and on recovery from oxidative stress due to a high-fat diet.
    Methods
    30 mice were divided into three groups: group 1 received a normal diet, group 2 received a high-fat diet, and group 3 received a high-fat diet plus 30 mg/kg pioglitazone. After treatment, the cholesterol, triglyceride, paraoxonase 1 (PON1), total serum antioxidant capacity (TAC), malondialdehyde (MDA), and specific activity of hepatic catalase were measured. BAT UCP1 expression was evaluated at both the mRNA and protein levels.
    Results
    The weights differed between the groups (p<0.05). Serum MDA was greater and TAC, liver catalase, and PON1 were less than in group 2 than in group 1 (p<0.05). In Serum MDA was less and catalase activity was greater in group 3 than in group 2 (p<0.05). UCP1 gene expression was less in group 2 than in group 1 (p<0.05) but greater than in group 3 (p<0.05).
    Conclusions
    Pioglitazone may have a protective role in high-fat-diet-induced oxidative stress by increasing the antioxidant capacity. Moreover, it can induce weight loss by increasing UCP1 mRNA and protein expression.
    Keywords: High-fat diet, MDA, pioglitazone, PON1, TAC, UCP1
  • Mojdeh Akbari, Atieh Abedin Do, Fakhrolmolouk Yassaee, Reza Mirfakhraie* Pages 21-24
    Background
    Uterine leiomyoma, also called fibroid, is a benign tumor that arises due to monoclonal transformation of myometrium, the smooth muscle cell layer of the uterus. Fibroids cause several complications including infertility, miscarriage, bleeding, pain, and dysmenorrhea. Recent studies have revealed the role of mutations in MED12 gene exon 2 in various populations; however, the reported frequency of these mutations differs between reports. In addition, it is suggested that mutations in exon 1 may also play a role in leiomyoma. The aim of the present study was to screen for MED12 exon 1 mutations in leiomyoma tissue samples of Iranian patients.                                                  .
    Methods
    We performed mutational analysis of exon 1 and the flanking intronic regions using multi-temperature single-strand conformational polymorphism (MSSCP) and sequencing analyses in 120 uterine leiomyoma samples.
    Results
    No mutations were detected in exon 1 of MED12 in our samples.
    Conclusions
    According to the literature and the present results, mutations in the MED12 exon 1 are rare. However, we could not ignore the role of these mutations in developing leiomyoma.
    Keywords: Exon 1, MED12, Mutation, Uterine leiomyoma
  • Iman Azari, Soudeh GhafouriFard, Mir Davood Omrani, Shahram ArsangJang, Dor Mohammad Kordi Tamandani, Mehrnaz Saroone Rigi, Sara Rafiee, Farkhondeh Pouresmaeili, Mohammad Taheri* Pages 25-31
    Background
    Intrauterine growth restriction (IUGR), a pathologic diminution of the rate of fetal growth, has been associated with alterations in expression of several genes. However, the role of long non-coding RNAs (lncRNAs) in its pathogenesis has not been studied.
    Methods
    In this study we evaluated the expression of four lncRNAs namely, nuclear paraspeckle assembly transcript (NEAT1), taurine up-regulated 1 (TUG1), p21-associated ncRNA DNA damage-activated (PANDA), and metastasis-associated lung adenocarcinoma transcript-1 (MALAT1) in placenta samples obtained from IUGR and normal pregnancies to determine their possible contributions in the pathogenesis of IUGR.
    Results
    We found no significant differences in expression levels between cases and controls. We also found no correlation between expression and clinical data of study participants; however, we found significant correlations between expression levels of all the assessed lncRNAs in both cases and controls.
    Conclusions
    These results imply the existence of a possible shared regulatory mechanism for the expression of these transcripts in placenta. Future studies are needed to perform such evaluations in larger sample sizes or in animal models in earlier stages of pregnancy.
    Keywords: IUGR, NEAT1, MALAT1, PANDA, Placenta, TUG1
  • Faria Hasanzadeh Haghighi, Ehsan Aryan, Aida Gholoobi, Hosna Zare, Zahra Meshkat* Pages 32-35
    Background
    Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and especially prevalent in developing countries. Several vaccines against TB have been developed, recently. The aim of the present study was to design and construct a cloning vector encoding Mycobacterium tuberculosis (MTB) mpt51 gene.
    Methods
    DNA was extracted from MTB H37Rv strain. Gene-specific primers were designed using Gene Runner software and the mpt51 gene was amplified by PCR. The amplified fragment and pcDNA3.1(+) cloning vector were both digested with restriction enzymes, the mpt51 fragment was ligated into the vector, and the Escherichia coli (E. coli) TOP10 strain were transformed by the recombinant plasmid. Positive clones were identified by colony PCR, restriction enzyme digestion, and DNA sequencing.
    Results
    The mpt51 gene was successfully cloned into pcDNA3.1(+). A 6400 bp band for the pcDNA3.1(+)/mpt51 recombinant plasmid and a 926 bp band for mpt51 were observed by colony PCR, and restriction enzyme digestion on agarose gels. The DNA sequence was 100% homologous with the mpt51 fragment of H37Rv in GenBank.
    Conclusions
    In the current study, the mpt51 gene of MTB was correctly cloned into pcDNA3.1(+). The expression of this recombinant vector can be studied in eukaryotic cells. Moreover, it is possible to determine the efficacy of this vector as a DNA vaccine candidate, and to test its protective function compared to BCG in animal models in future.
    Keywords: Molecular Cloning, MPT51antigen, Mpt51gene, Mycobacterium tuberculosis
  • Farbod Esfandi, Hamid Fallah, Shahram ArsangJang, Mohammad Taheri*, Soudeh GhafouriFard Pages 36-41
    Background
    Long non-coding RNAs (lncRNAs) have been considered to be prospective biomarkers for diagnosing lung cancer due to the fundamental roles they hold in the regulating several cancer-related pathways. 
    Methods
    Using the quantitative real-time polymerase chain reaction method, we evaluated the expression of CCAT2, UCA1, PANDA and GHET1 lncRNAs in 32 lung cancer tissue samples and their corresponding adjacent non-cancerous tissues (ANCTs) from lung cancer patients admitted to the Labbafi-Nejad Hospital from 2015 to 2016.
    Results
    No significant differences were found in the expression of lncRNAs within the tumoral and non-tumoral tissue samples. Bayesian Multilevel analysis showed no association between the expression of lncRNAs and the patient’s tumor node metastasis (TNM) stage following adjustments for age. Spearman correlation analysis revealed an inverse correlation between the expression of PANDA in tumoral tissues and age. Additionally, the difference in CCAT2 expression among the tumoral and non-tumoral tissues was inversely correlated with patients' age. Significant pairwise correlations were found between the expression of lncRNAs in both the tumoral and non-tumoral tissues.
    Conclusions
    Despite the findings supporting a role for the lncRNAs, CCAT2, UCA1, PANDA and GHET1 in the pathogenesis of lung cancer, our data suggests no relationship for expression of these lncRNAs in lung cancer, questioning their potential as lung cancer biomarkers.
    Keywords: CCAT2, GHET1, lncRNAs, Lung cancer, PANDA, UCA1
  • Hassan Ahmadvand, Banafsheh Yalameha, Glavizh Adibhesami, Maryam Nasri, Negar Naderi, Esmaeel Babaeenezhad, Negar Nouryazdan* Pages 42-48
    Background
    Renal ischemia-reperfusion injury (RIR) occurs when there is a temporary restriction of blood flow to the kidneys followed by an influx of blood, re-oxygenating the tissues. This occurs as a severe complication of major surgery. This process causes significant damage to the tissues and is responsible for the development of acute kidney injury (AKI), a life-threatening condition with high mortality rates. Here, we evaluated the potential protective effects of the antioxidant, gallic acid (GA), on RIR in an in vivo rat model.
    Methods
    Adult male Sprague Dawley rats were randomly divided into three groups: group 1 (control, n = 8), group 2 (Ischemia-reperfusion (IR) with no-treatment, n = 7), and group 3 (IR + daily GA 100 mg/kg i.p, n = 7). The abdomens of the rats in the control group were opened during the surgical procedure, then sutured closed. GA pretreatment began daily 15 days prior to inducing RIR. To induce RIR, the umbilical arteries were obstructed on both sides and clamped with mild pressure for 45 min. Following the 45 min ischemia, the clamps were removed to allow for the induction of reperfusion. The reperfusion phase was 24 hours.
    Results
    Following IR, the serum levels of urea and creatinine significantly increased compared to the controls. Pretreatment with GA was observed to reduce urea and creatinine levels following IR. However, this decrease was not statistically significant. The serum and renal levels of malondialdehyde (MDA) in the IR group was significantly elevated compared to the control group. Conversely, glutathione (GSH) levels and the activity of glutathione peroxidase (GPX) significantly decreased in the IR group compared to controls. Our findings show GA pretreatment to significantly improve the levels of renal MDA, serum GSH, and GPX activity following RIR.
    Conclusions
    Our findings highlight the protective role for GA in mitigating the damage caused by RIR and its applications as a potential treatment.
    Keywords: Antioxidant enzymes, Gallic acid, Renal functional markers, Renal ischemia-reperfusion
  • Mostafa Cheraghi, Hassan Ahmadvand*, Ali Maleki, Esmaeel Babaeenezhad, Salman Shakiba, Fatemeh Hassanzadeh Pages 49-55
    Background
    Oxidative stress plays an important role in the development of atherosclerosis. An association exists between the alterations of liver markers and the risk of coronary heart disease (CHD). This study was designed to investigate the status of oxidative stress and liver markers in patients with CHD.
    Methods
    This study included 50 CHD patients and 50 healthy volunteers. Serum activities of glutathione peroxidase (GPX), catalase (CAT), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), and fasting blood sugar (FBS) concentrations were measured. The Unpaired Student’s t-test was used to analyze the data.
    Results
    Serum GSH level and CAT and GPX activities were significantly greater in healthy controls than in CHD patients. Serum MDA, NO, and FBS levels and GGT, ALT, ALP activities were significantly greater in CHD patients than in healthy controls. Serum AST activity was greater in CHD patients than in controls, but the difference was not statistically significant.
    Conclusions
    Our results indicate that CHD is related to oxidative stress, lipid peroxidation, inflammation, and elevated liver enzyme activity. CHD is a deadly disease that requires appropriate medical care. Antioxidant treatment might inhibit disease progression.
    Keywords: Coronary heart disease, Inflammation, Liver markers, Oxidative stress
  • Arezou Sayad, Soudeh GhafouriFard, Rezvan Noroozi, Mir Davood Omrani, Maziar Ganji, Romina Dastmalchi, Mark Glassy, Mohammad Taheri* Pages 56-62
    Background
    Autism spectrum disorders (ASDs) (MIM 209850) are a group of distinct neurodevelopmental disorders characterized by impaired social interactions and communication abilities and abnormal repetitive activities. Many genetic variants have been shown to be associated with ASD. Channelopathies are among putative culprits in the pathogenesis of many neurodevelopmental disorders, including autism. The calcium channel, voltage-dependent, L type, alpha 1C subunit gene (CACNA1C) encodes an alpha-1 subunit of a voltage-dependent calcium channel. Genetic variants within this gene have been associated with psychiatric disorders including Autism Spectrum Disorders (ASD). Our aim was to determine whether the SNPs rs1006737, rs4765905, and rs4765913 were associated with ASD in an Iranian population. 
    Methods
    In the present case-control study we investigated the associations of rs1006737, rs4765905, and rs4765913 polymorphisms within CACNA1C and the risk of ASD in a population of 529 Iranian ASD patients and 480 age, gender, and ethnicity-matched healthy subjects.
    Results
    None of these SNPs were associated with ASD risk in the assessed population. Although previous studies have shown an association between these polymorphisms and psychiatric disorders and an association between rs4765905 and ASD, we did not replicate those results in our study.
    Conclusions
    Our data indicate that these CACNA1C variants are not involved in the pathogenesis of ASD in the Iranian population.
    Keywords: Autism Spectrum Disorder, CACNA1C, Channelopathy, polymorphism
  • Ladan Kabiri, Shiva Irani, Amin Reza Nikpoor, Jalil Tavakkol Afshari* Pages 63-71
    Background
    Many studies have shown the anticancer effects of mannan and mitomycin C on tumor cells. In this regard, the aim of this study was to investigate the inhibitory and apoptotic effects of a mannan-mitomycin-C conjugate on transitional cell carcinoma (TCC) and normal mouse L929 fibroblast cells.
    Methods
    The conjugate was synthesized according to previous studies. Both cell lines were cultured and the cytotoxic and apoptotic effects of the compounds in different concentrations were assessed using MTT and flow cytometry, respectively. The mannan-mitomycin C conjugate inhibited proliferation of both cell lines in time and concentration -dependent manners.
    Results
    The conjugate inhibited TCC cell proliferation more than that of L929 cells. Mitomycin C alone inhibited proliferation of both cell lines in both time and concentration -dependent manners, and the effect was greater on L929 than on TCC cells. Mannan had a relatively low inhibitory effect on TCC and no significant effect on L929 cells. The percentage of apoptosis was greater in TCCs than in L929 cells at the highest concentration of conjugate. Mitomycin C induced apoptosis more extensively in L929 cells than in TCC cells at 25 and 400 μg/ml. The effect of mannan was similar on both cell lines.
    Conclusions
    The mannan-mitomycin C conjugate has greater inhibitory and apoptotic effects on TCC than on L929 cells and may inhibit TCC.
    Keywords: Apoptosis, Inhibitory Effect, Mannan-Mitomycin C Conjugate, MTT, TCC
  • Chanda Jha, Shobha Ullas Kamath*, Sambit Dash, Ravindra Prabhu Attur, Lingadakai Ramachandra, Rajgopal Shenoy Kallya Pages 72-78
    Background
    Following contrast-enhanced computed tomography (CECT) contrast-induced nephropathy (CIN) may occur in patients with renal insufficiency or diabetes. Creatinine, the most common marker of CIN, may not be an accurate measure of damage and is affected by many non-renal factors. Our aim was to evaluate ischemia-modified albumin (IMA) as an early CIN marker and correlate it with paraoxonase-1 (PON-1) and creatinine before and after CECT.
    Methods
    Forty-eight adult patients scheduled for intravenous CECT, regardless of indication or body region for CECT, were included in this prospective study. Venous blood samples were obtained 12-24 hours before and after contrast media (CM) administration. Ischemia-modified albumin and PON-1 were estimated using methods described by Bar-Or et al. and Dantoine et.al., respectively. Creatinine was estimated on an automated analyzer.
    Results
    Significant differences in IMA (P < 0.001) and PON-1 (P < 0.001) levels were found between pre- and post-CECT samples, while the difference for creatinine was not significant (p = 0.073). No correlation was found between IMA and PON-1 or IMA and creatinine in either the pre- or post-CECT samples.
    Conclusions
    After CM administration patients are subjected to oxidative stress and/or ischemia, as revealed by elevated IMA and decreased PON-1 levels; however, creatinine levels, most commonly estimated to assess reduced renal function, did not reflect the condition accurately. IMA may be a sensitive marker for CIN but further studies are required to confirm its usefulness.
    Keywords: Contrast media (CM), Contrast-enhanced computed tomography (CECT), Contrast-induced nephropathy (CIN), Creatinine, Ischemia-modified albumin (IMA), Paraoxonase-1 (PON-1)
  • Mustafa Mansouri, Seyed Adel Moallem, Javad Asili, Leila Etemad* Pages 79-84
    Background
    Breast cancer is the most prevalent cancer in women worldwide, especially in developing countries. Scrophularia umbrosa Dumort, a medicinal plant, has been used to treat various diseases in traditional medicine. In this study, we investigated the anti-cancer and cytotoxic effects of S. umbrosa Dumort extracts on a human breast cancer cell line.
    Methods
    The methanol and other S. umbrosa Dumort factions, including those from dichloromethane, water, n-butanol, ethyl acetate, and petroleum ether, were examined. The cytotoxic effects of the fractions on MCF-7 human breast cancer adenocarcinoma and 3T3 mouse embryonic fibroblast cells were evaluated by MTT assays. In addition, apoptotic induction was determined by propidium iodide flow cytometry.
    Results
    The water, n-butanol. petroleum ether, and ethyl acetate fractions had no cytotoxic effects. The methanol and dichloromethane fractions showed significant cytotoxic affects in a dose-dependent manner on the malignant cells while causing no damage to non-malignant cells. In addition, the cell death assay indicated that the S. umbrosa dichloromethane fraction triggered apoptosis in the MCF-7 cells.
    Conclusions
    S. umbrosa induced apoptosis in MCF-7 cells. The S. umbrosa dichloromethane fraction exhibited the greatest cytotoxic effect on these cells. This work presents a first evaluation of the cytotoxic effects of S. umbrosa and further studies are needed to determine the cytotoxic mechanism.
    Keywords: Apoptosis, Breast cancer, Cytotoxicity, MCF-7 cell line, Scrophularia umbrosaDumort
  • Fatemeh Talebi, Mehrnaz Rasooli Nejad, Mehdi Yaseri, Azar Hadadi* Pages 85-90
    Background
    Community-acquired pneumonia (CAP) is a common disease considered as a major public health problem. It causes considerable morbidity and mortality despite antibiotic treatments. Hospital admission of CAP patients is a significant financial burden and many efforts are ongoing to decrease hospital stay durations. Vitamin D deficiency is associated with increased risk of respiratory infections. This study was designed to determine the association of vitamin D status with hospitalized CAP patient mortality and disease severity.
    Methods
    This prospective cohort study examined 180 CAP patients admitted to a teaching Hospital in Tehran, Iran during 2016-2017. Their demographic and anthropometric characteristics were recorded. The disease severity was evaluated based on CURB-65. Vitamin D status was determined by measuring by serum 25-hydroxylated vitamin D (25(OH)D) with ELISA. The patients were followed for 30 days to evaluate their vitality.
    Results
    One hundred and eighty pneumonia patients, including 104 males and 84 females, were recruited from respiratory disease, infectious disease, emergency, and ICU wards. Nearly 18% of the patients were current smokers. The CAP severity, evaluated by CURB-65, was determined to be non-severe in 74.4% of the patients. Patients were classified as vitamin D sufficient, insufficient, or deficient. Thirty percent of the patients were vitamin D sufficient, 18% were insufficient, and 52% were deficient. Thirty-day mortality was 40% (72 cases).
    Mortality was greater in males than in females (47.1% vs. 30.3%, p=0.03). The disease was significantly less severe in the patients who survived than in those who did not. The vitamin D status differed between males and females (p=0.027). The vitamin D status was lower in the more severe cases than in the less (p=0.036), and vitamin D deficiency was more prevalent in patients who died than in those who lived. Vitamin D concentration was negatively correlated with hospital stay duration. The 25(OH)D concentration was significantly greater in patients who survived than in those who did not (p<0.001).
    Conclusions
    Pneumonia severity and mortality risk were greater and hospital stays longer in vitamin D-deficient patients than in those with higher vitamin D status.
    Keywords: Disease severity, Mortality, Pneumonia, Vitamin D
  • Mohsen Soosanabadi, Reza Mirfakhraie, Lilit Atanesyan, Akbar Biglarian, Fatemeh Aghakhani Moghadam, Maryam Rahimi, Farkhondeh Behjati, Elaheh Keyhani* Pages 91-101
    Background
    The aim of this study was to assess the usability of multiplex ligation-dependent probe amplification (MLPA) for copy number determination of HER gene family members (ERBB1-4) in invasive breast carcinoma and to explore the association of ERBB1-4 gene copy numbers with clinicopathological characteristics of breast cancer (BC) patients.
    Methods
    Clinical and immunohistochemical characteristics were assessed in 104 BC patients and the molecular subtype was determined for each tumor sample. Furthermore, HER-2/neu status was assessed by immunohistochemistry (IHC) and equivocal results were confirmed by Fluorescent in situ hybridization (FISH). The copy numbers of ERBB1-4 genes were determined by MLPA.
    Results
    Twenty-five percent of all patients showed ERBB2 gene-amplification by MLPA, whereas 14.4% of cases showed ERBB-2/neu overproduction at the protein level (IHC). Moreover, only 2.9% and 1.9% of patients showed amplification in ERBB1 and ERBB4, respectively. No copy number changes were observed in ERBB3. Our results indicated a significant association between ERBB2 copy number gain and histological grade (p value= 0.01), stage (p value= 0.02), and tumor subtypes (p value= <0.001). In addition, we found MLPA more accurate in assessing HER2 status with 15.4% and 9.6% gene amplification detection in early stages (1, 2A and 2B) and advanced tumor stages (3A, 3B, and 4), respectively, compared to IHC (early stages= 13.5% and advanced stages= 4.7%).
    Conclusions
    According to our findings, MLPA is a fast, precise and low-cost technique to detect ERBB2 amplification, especially in advanced tumor stages. However, due to infrequent amplification found in ERBB1 and ERBB4 as well as the lack of amplification in ERBB3, their importance in the prognostic evaluation of BC patients remains controversial.
    Keywords: Breast Cancer, Copy number variation, ERBB1-4, IHC, MLPA