Differential Expression of miR-143-5p and miR-744-5p with HIF1A Across Breast Cancer Grade and Treatment status in Iraqi Females

Breast cancer is a malignant tumor that originates in breast tissue and the most common cause of malignant tumors-related death in women worldwide. Long noncoding RNA (LncRNA) is a transcribed RNA with more than 200 nucleotides in length but has no notable potential for protein coding. Hypoxia is a common occurrence in tumor microenvironments due to an imbalance in oxygen supply and consumption by rapidly growing tumors. The aim of study is to analyze the gene expression of different types of lncRNAs (lnc.4.2, lnc.4.3) as a molecular tumor marker in relation with hypoxia-inducible factor 1alpha (HIF-1α) gene and main clinic pathological characters for patients and healthy control group. The study included 100 newly diagnosed cases of bc divided into four groups: (n = 25) low grade before treatment (LBT), (n = 25) low grade after treatment (LAT), (n = 25) high grade before treatment (HBT), and (n = 25) high grade after treatment (HAT). In addition, (n = 25) as a control group. The results of this study showed that lncRNA 4.2 and lncRNA 4.3 exhibited strong discriminatory ability specifically in the LBT versus control and HAT versus control comparisons, with AUC values exceeding 0.70. This indicates that these lncRNAs may be particularly useful for distinguishing LBT and HAT samples indicate potential role in treatment response or the pretreatment state. HIF1A also exhibited moderate diagnostic potential in the HBT versus control and HAT versus control comparisons, with AUC values greater than 0.60. Its performance was notably better in the HBT group, where it achieved an AUC of 0.774, indicating good discriminatory ability in this specific context. This suggests that HIF1A may be a useful diagnostic marker for conditions associated with HBT samples, potentially reflecting its role in cellular stress response.

Background: worldwide, breast cancer is the most common malignancy among women and there is a deep need for precise novel methodologies for breast cancer (BC) diagnosis. Major advances in cancer control will be successfully achieved with early cancer detection. So, recent trends are going toward using circulating non-coding RNA as diagnostic tool for their critical role in cancer detection. Aim: retrieve non coding RNA that is mechanistically linked to breast cancer stem cell with validation of the results in a group of breast cancer patients versus control groups to evaluate their usefulness as a potential biomarker in breast cancer diagnosis. Patients and Methods: we retrieved LncRNA that is linked to stem cell differentiation and specific to BC utilizing bioinformatics tools. Then we validated this biomarker in serum of 30 patients with BC, 12 patients with benign breast lesion and 12 healthy volunteers using RT-qPCR. We evaluate the power of diagnosis of the serum profiling system using ROC curve analysis. Results: hoxd antisense growth-associated long non coding RNA (HAGLR) had great sensitivity and specificity for differentiating BC from patients with benign breast lesion and also from healthy controls. Conclusion: the chosencirculatory RNA based biomarker can be used as a potential diagnostic biomarker for BC. In addition it could be therapeutic target.

Background: worldwide, breast cancer is the most common malignancy among women and there is a deep need for precise novel methodologies for breast cancer (BC) diagnosis. Major advances in cancer control will be successfully achieved with early cancer detection. So, recent trends are going toward using circulating non-coding RNA as diagnostic tool for their critical role in cancer detection. Aim: retrieve non coding RNA that is mechanistically linked to breast cancer stem cell with validation of the results in a group of breast cancer patients versus control groups to evaluate their usefulness as a potential biomarker in breast cancer diagnosis. Patients and Methods: we retrieved LncRNA that is linked to stem cell differentiation and specific to BC utilizing bioinformatics tools. Then we validated this biomarker in serum of 30 patients with BC, 12 patients with benign breast lesion and 12 healthy volunteers using RT-qPCR. We evaluate the power of diagnosis of the serum profiling system using ROC curve analysis. Results: hoxd antisense growth-associated long non coding RNA (HAGLR) had great sensitivity and specificity for differentiating BC from patients with benign breast lesion and also from healthy controls. Conclusion: the chosencirculatory RNA based biomarker can be used as a potential diagnostic biomarker for BC. In addition it could be therapeutic target.

We performed genome-wide characterization of Han Chinese breast cancer at molecular level by integrating two microarray technologies: comparative genomic hybridization (CGH) for DNA copy number variations (CNV) and expression arrays. Concurrent gains and losses from the same subject across genomic and transcriptional levels may be a better approach to explore potential biomarkers for breast cancer and to identify possible candidates for target therapy. Oncogenesis of breast cancer could originate from chromosome level manifesting as CNV and persist through transcription in gene expression profiles. Genes with coherent patterns at chromosomal and transcriptional level are more likely to serve as potential biomarkers for Han Chinese breast cancer and deserve meticulous evaluation. A total of 23 array CGH and 81 gene expression microarrays (21 samples with data from both platforms) were performed. Potential targets of cancer therapy were revealed by Genomic Identification of Significant Targets in Cancer (GISTIC) from 23 array CGH. Genes with coherent patterns between CNV and gene expression profiles were identified from 21 samples with both platforms assayed. We used these concurrent genes as well as genes with significant GISTIC scores to derive signatures associated with ER, HER2 and event-free survival. Distributions of signature genes were strongly associated with chromosomal locations: chromosome 16 for ER and 17 for HER2. 37 and 13 genes were differentially expressed between distinct ER and HER2 status and were used for classifier development in our 81 samples. Predictive accuracy as high as 90% for ER and 80% for HER2 was reported during leave-one-out cross-validation while much compromised performance was observed in an independent test dataset (GSE5460). Consensus genes between our dataset and 125 Chinese breast cancers (GSE5460) were identified, and a 28 genes and a 9 genes signature for ER and HER2 was developed in combined dataset of 206 microarrays with the best predictive accuracy of 92% and 94% for ER and HER2 respectively. Breast cancer risk predictive model was built based on supervised principle components from two genes (RWDD3 and ZBTB44) and distinct survival patterns were observed between high- and low-risk groups from a combined dataset of 408 microarrays (combination of our 81 samples with GSE20685). The risk score was significantly higher in breast cancers with recurrence, metastasis or mortality than those remained event-free (0.438 versus 0.17, p<0.001). In summary, we built a breast cancer risk predictive model and distinct survival patterns were observed in Han Chinese breast cancers. Citation Format: Chi-Cheng Huang, Shih-Hsin Tu, Eric Y. Chuang. Concurrent genes signatures for breast cancers with Han Chinese origin. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4035. doi:10.1158/1538-7445.AM2013-4035

Breast cancer (BC) is one of the most prevalent malignancies among women worldwide, and early detection is crucial for improving patient outcomes. Among the key signaling pathways involved in BC, the MAPK signaling pathway plays a significant role in tumor progression. Through bioinformatics analysis, we identified miRNAs associated with the MAPK signaling pathway. Differential expression analysis using microarray data from the GSE106817 and GSE113486 datasets further refined our selection. Based on these analyses, we identified two candidate miRNAs, hsa-miR-27a-5p and hsa-miR-449a, as potential biomarkers for BC. We then examined the expression levels of these two serum miRNAs in women with BC (n = 100) and healthy controls (n = 100) using RT-PCR. Our results demonstrated that both miRNAs were significantly upregulated in women with BC compared to healthy individuals (p < 0.05). Receiver operating characteristic (ROC) curve analysis further confirmed that these miRNAs exhibit strong diagnostic potential for BC. Overall, our findings suggest that hsa-miR-27a-5p and hsa-miR-449a, identified through bioinformatics and microarray-based expression analyses, could serve as promising biomarkers for BC detection. Their significant association with the MAPK signaling pathway was determined through bioinformatics analysis.

Study of microRNAs-21/221 as potential breast cancer biomarkers in Egyptian women

Recent advances suggest that abnormal fatty acid metabolism highly correlates with breast cancer, which provide clues to discover potential biomarkers of breast cancer. This study aims to identify serum free fatty acid (FFA) metabolic profiles and screen potential biomarkers for breast cancer diagnosis. Gas chromatography-mass spectrometry and our in-house fatty acid methyl ester standard substances library were combined to accurately identify FFA profiles in serum samples of breast cancer patients and breast adenosis patients (as controls). Potential biomarkers were screened by applying statistical analysis. A total of 18 FFAs were accurately identified in serum sample. Two groups of patients were correctly discriminated by the orthogonal partial least squares-discriminant analysis model based on FFA profiles. Seven FFA levels were significantly higher in serum from breast cancer patients than that in controls, and exhibited positive correlation with malignant degrees of disease. Furthermore, five candidates (palmitic acid, oleic acid, cis-8,11,14-eicosatrienoic acid, docosanoic acid and the ratio of oleic acid to stearic acid) were selected as potential serum biomarkers for differential diagnosis of breast cancer. Our study will help to reveal the metabolic signature of FFAs in breast cancer patients, and provides valuable information for facilitating clinical noninvasive diagnosis.

Athletic performance is a polygenic trait influenced by both environmental and genetic factors. 1 Genetics has a great influence over athletic performance such as strength, power, muscle fibre, flexibility and other phenotypes. HIF1A (hypoxia inducible factor 1 alpha), a master-regulator of oxygen-homeostasis, plays crucial roles in cellular metabolism. 2 HIF1A regulates expression of nearly all enzymes involved in glycolysis. HIF1A mediates oxygen delivery to cells as well as allows cells to survive under oxygen deprivation by regulating expression of hundreds of genes involved in angiogenesis, erythropoiesis glucose metabolism and transport. 2,3 The HIF1A gene is considered as a genetic marker of athletic ability because of its proposed role in shifting of oxidative type muscle fibres to glycolytic type and increasing hypoxia resistance to the cells. 4 The aim of this study was to perform a meta-analysis of all available case-control-studies of athletes to systematically summarise the association between HIF1A C1772T (rs11549465) polymorphism and athletic performance. Literature-mining was performed for the last 15 years using PubMed and Google Scholar to retrieve relevant case-control studies. Odds ratio (OR) with 95% confidence intervals (CIs) were calculated using fixed or random effects model incorporating inverse-variance-weighted method for relating HIF1A 1772 C/T polymorphism (rs11549465; which results in Pro582Ser substitutions) with elite athlete’s performance. In subgroup analyses top-level athletes were considered as ‘elite athletes’ who were competing in the Olympics Games &/World and European championships and won medals; and ‘other athletes’ being national level and regional competitors with no less than four years of experience participating in their sport. The distributions of genotypes in the controls were checked for HWE. Further subgroup-analysis of HIF1A 1772 C/T was carried out by ethnicity. A meta-analysis of 19 studies comprising 1616 cases and 9253 controls was conducted to identify the association of HIF1A rs11549465 (C/T) polymorphism with elite athlete’s performance. Results demonstrated that the T allele is significantly associated with elite athletes under three genetic models (TT+CT vs CC: OR = 1.73, 95% CI: = 1.22–2.46, p 0.002); (CT vs CC: OR = 1.73, 95% CI: = 1.21–2.47, p 0.003); (T vs C: OR = 1.64, 95% CI: = 1.19–2.25, p 0.002) whereas other athletes group showed no significant associations (TT+CT vs CC: OR = 1.44, 95% CI: = 0.72–2.88, p 0.30); (CT vs CC: OR = 1.37, 95% CI: = 0.65–2.89, p 0.40); (T vs C: OR = 1.38, 95% CI: = 0.84–2.29, p 0.20). For elite athletes no significant heterogeneity was observed (T vs C: I^2 = 60.8% [0% - 86.9%]; Q = 7.6; p 0.054). For Caucasian population, sub-group analyses data suggested that the HIF1A 1772 C/T polymorphism is highly associated with athletic performance in three models (TT+CT vs CC: OR = 1.51, 95% CI: = 1.15–1.98, p 0.003); (CT vs CC: OR = 1.53, 95% CI: = 1.17–2.01, p 0.002); (T vs C: OR = 1.41, 95% CI: = 1.1–1.80, p 0.005). Results from this study revealed that HIF1A C1772T polymorphism is significantly associated with elite athletes compared with controls or other athletes. This suggests that the T allele variant could be one of the factors influencing the power athletic performance. Acknowledgment This work is supported by the research & development allocation Grant: Ministry of Science & Technology (MOST), Government of the People’s Republic of Bangladesh [39.009.002.01.00.057.2015-2016/1425] and the special allocation Grant: Ministry of Education, Government of the People’s Republic of Bangladesh [37.20.0000.004.033.013.2015]. References Drozdovska SB, Dosenko VE, Ahmetov II.,and Ilyin VN. The association of gene polymorphisms with athlete status in Ukrainians. Biol Sport . 2013; 30 (3):163–167. Ke Q, Costa M. Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol . 2006; 70 :1469–80. Clifford SC, Astuti D, Hooper L, Maxwell PH, Ratcliffe PJ, Maher ER. The pVHL-associated SCF ubiquitin ligase complex: molecular genetic analysis of elongin B and C, Rbx1 and HIF-1alpha in renal cell carcinoma. Oncogene . 2001; 20 (36):5067–74. PMID 11526493 Cieszczyk P, Eider J, Arczewska A, Ostanek M, Leonska-Duniec A, Sawczyn S, Ficek K, Jascaniene N, Kotarska K and Sygit K. The HIF1A gene Pro582Ser polymorphism in Polish power-oriented athletes. Biol. Sport 2011; 28 :111–114.

IntroductionBreast cancer (BC) remains the most prevalent malignant tumor in women worldwide and a leading cause of cancer-related mortality. Early screening is essential to improve prognosis, yet current diagnostic methods are often invasive or lack sensitivity. Saliva is an accessible and non-invasive biofluid containing various metabolites that reflect systemic physiological and pathological changes. Thus, salivary metabolomics may provide novel insights into breast cancer-associated metabolic alterations and support the development of early diagnostic strategies.ObjectivesTo explore the salivary metabolomic profile of breast cancer patients and identify potential non-invasive biomarkers for early breast cancer screening.MethodsSaliva samples were collected from a screening set consisting of 30 BC patients and 20 normal controls (NC) volunteers. untargeted metabolomics approach was performed using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), along with KEGG pathway enrichment and receiver operating characteristic (ROC) curve analyses, were employed to characterize metabolic differences and identify potential biomarkers. Additionally, saliva samples from a validation set (52 BC patients and 52 NC volunteers) were collected. Enzyme-linked immunosorbent assay (ELISA) was used to quantify potential biomarkers, and their diagnostic performance was evaluated through ROC curve analysis.ResultsA total of 101 differential metabolites were identified, including 81 upregulated and 20 downregulated compounds. Screening identified 2-aminonicotinic acid and theobromine as potential diagnostic biomarkers. Analysis of the validation set demonstrated that 2-aminonicotinic acid (AUC: 0.81, cut-off: 5.88 ng/mL) and theobromine (AUC: 0.75, cut-off: 5.27 ng/mL) exhibit promising diagnostic potential.ConclusionThe salivary metabolome of breast cancer patients displays distinct changes compared to healthy individuals. Salivary 2-aminonicotinic acid and theobromine emerge as promising non-invasive biomarkers for breast cancer detection. Nevertheless, larger-scale validation studies are warranted to substantiate their specificity and clinical utility.

Identification of Differentially Expressed Plasma lncRNAs As Potential Biomarkers for Breast Cancer

Association of hypoxia inducible factor-1α polymorphisms with susceptibility to non–small-cell lung cancer

Objectives:To investigate the circulating levels of microRNA-34a (miRNA-34a) as a novel non-invasive biomarker of breast cancer (BC).Methods:The case-control study was conducted at the Department of Chemistry and Biochemistry, College of Medicine, Al-Nahrain University, Baghdad, Iraq, from December 2018 to April 2019. Real-time quantitative polymerase chain reaction has been employed to analyze miRNA-34a expression in the samples of serum from 90 participants (30 patients with BC, 30 patients with benign breast tumors, and 30 control subjects) after RNA extraction and reverse transcription. Cancer antigen 15-3 (CA15-3) and carcinoembryonic antigen (CEA) were measured by ELISA. Additionally, we analyzed the receiver operating characteristic curves of various markers, including miRNA -34a, CA15-3, and CEA, to assess the diagnostic power of each marker.Results:The expression of miRNA-34a has been significantly lower in the group of breast cancer compared with that in the group of control, and miRNA-34a expression has been significantly reduced in the group of benign breast tumor compared as that in the group of control. Receiver operating characteristics analysis showed a very good discriminative power of combined miRNA-34a and CA15-3 (specificity=77.7%; sensitivity=83.3% and areas under the curve =0.842) for BC patients.Conclusion:MicroRNA-34a expression is significantly decreased in the patients’ serum with the cancer of breast, and miRNA-34a can be employed as a potential non-invasive molecular marker for the early diagnosis of BC.

Disruption of telomere length (TL) homeostasis in peripheral blood lymphocytes has been previously assessed as a potential biomarker of breast cancer (BC) risk. The present study addressed the relationship between lymphocyte TL (LTL), prognosis and clinicopathological features in the BC patients since these associations are insufficiently explored at present. LTL was measured in 611 BC patients and 154 healthy controls using the monochrome multiplex quantitative Polymerase Chain Reaction assay. In addition, we genotyped nine TL-associated single-nucleotide polymorphisms that had been identified through genome-wide association studies. Our results showed that the patients had significantly (P = 0.001, Mann-Whitney U-test) longer LTL [median (interquartile range); 1.48 (1.22-1.78)] than the healthy controls [1.27 (0.97-1.82)]. Patients homozygous (CC) for the common allele of hTERT rs2736108 or the variant allele (CC) of hTERC rs16847897 had longer LTL. The latter association remained statistically significant in the recessive genetic model after the Bonferroni correction (P = 0.004, Wilcoxon two-sample test). We observed no association between LTL and overall survival or relapse-free survival of the patients. LTL did not correlate with cancer staging based on Union for International Cancer Control (UICC), The tumor node metastasis (TNM) staging system classification, tumour grade or molecular BC subtypes. Overall, we observed an association between long LTL and BC disease and an association of the hTERC rs16847897 CC genotype with increased LTL. However, no association between LTL, clinicopathological features and survival of the patients was found.

INTRODUCTION: Breast cancer is the most common malignancy in the female population and its early diagnosis is important for the success of treatment. Tumor cell-free DNA (cfDNA) circulating in plasma has been suggested as a new potential cancer biomarker. The aim of this study was to quantify the level of plasma cfDNA in plasma of patients with breast carcinoma and determine its correlation with prognostic factors for the disease outcome. MATERIAL AND METHODS: Plasma samples were obtained from 65 women with primary breast cancer and from 29 healthy female controls. Circulatory cfDNA was extracted from the samples and quantified by real-time quantitative PCR for HBB. RESULTS: The mean concentrations of cfDNA 32. 85 ng/ml-1 in breast cancer patients and 17. 08 ng/ml-1 in healthy controls (p=0. 02). Patients with metastatic disease had higher concentrations of cfDNA than patients without metastasis (p=0. 03). No correlation was found between cfDNA concentrations and breast cancer prognostic factors such as hormonal receptors and HER2 status. DISCUSSION AND CONCLUSION: We concluded that the levels of cfDNA are elevated in patients with breast carcinoma. These values are associated with the presence of metastases and advanced-stage tumors. The role of cfDNA as a potential biomarker in breast cancer is suggested.

LAPTM4B is a transmembrane that promotes autophagy and renders tumor cells resistant to metabolic and genotoxic stress. Increased expression of LAPTM4B has been found in breast, liver, lung, ovarian, uterine and gastric cancers. Overexpression of LAPTM4B contributes to chemotherapy resistance, proliferation and metastases. We analyze the expression of LAPTM4B in serum by ELISA method in a series of breast cancer patients in different clinical situations to assess its possible potential as a biomarker. A total of 135 patients were analyzed, 95 patients were in early breast cancer and 50 in metastatic disease. Of the 95 patients with early disease, 52 had a luminal A phenotype, 19 luminal B, 15 HER2 positive and 9 triple negative. And of the other 50 they were luminal in 34, HER2 positive in 11 and triple negative in 5. The attached table specifies the values of the LAPTM4B expression in the different clinical situations. GLOBAL LUMINAL A LUMNAL B HER2 TRIPLE NEGATIVE SITUATIONS Laptm4b n Laptm4b n Laptm4b n Laptm4b n Laptm4b n EARLY 9,6 ng/ml 95 7,7 ng/ml 52 12,8 ng/ml 19 13,4 ng/ml 15 7,1 ng/ml 9 METASTATIC 16,8 ng/ml 50 13,7 ng/ml 34 7,2 ng/ml 11 58,9 ng/ml 5 We found a statistic difference in the expression of LAPTM4B in patients with early and metastatic disease (ANOVA test p:0,028 ). LAPTM4B was higher in the luminal B and HER2 subtypes in early breast cancer while in metastatic disease was overexpressed in triple negative. An interesting finding was that in metastatic disease, LAPTM4B was overexpressed in patients with progression compared to patients in response. 30 ng/ml vs 10 ng/ml (ANOVA test p:0,011) LAPTM4B expression may be a potential biomarker in breast cancer since its expression is statistically different in early and metastatic disease (9,6 vs 16,8 ng/ml).In addition, patients with metastatic breast cancer who re in response also have lower levels of LAPTM4B compared with patients in progression. Monitoring the levels of LAPTM4B in serum can help us to predict the treatment response. Citation Format: Serafin Morales Murillo, Ariadna Gasol Cudós, Noemí Tuset Der-Abrain, Izaskun Urdanibia, Ana Velasco Sánchez. LYSOSOMAL-ASSOCIATED TRANSMENBRANE 4B (LAPTM4B) AS POTENTIAL BIOMARKER IN BREAST CANCER [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-24-07.