Investigating the Relationship between Human Breast Tumor Grading and Beta Estrogen Receptor Expression

Background: Breast cancer represents the most prevalent cancer among women and ranks as the third leading cause of cancer-related mortality in females. Given the critical role of early cancer detection, identifying biomarkers related to breast cancer is essential for prognosis and early identification. This study aims to explore the differential expression of the estrogen receptor-β (ESRβ) gene in cancerous tissue samples compared to standard tissue samples, aiming to identify a biomarker for predicting cancer prognosis.Method: This study collected 60 fresh tissue samples from breast cancer patients post-surgery, comprising 30 tumor samples and 30 standard samples adjacent to the tumor, which were immediately stored at -70 °C. Following RNA extraction and cDNA synthesis, the expression levels of the ESRβ gene and the Gapdh gene (as an internal control) were analyzed using the real-time polymerase chain reaction method.Results: The study examined the variation in ESRβ gene expression between tumor tissues and adjacent healthy tissues in breast cancer patients. An increase in gene expression was observed in tumor samples compared with the adjacent healthy tissue. Furthermore, statistical analysis using SPSS software revealed a significant correlation between the gene expression level and tumor grade across all examined groups. Additionally, the findings indicate a significant difference in gene expression between patients with and without tumor metastasis.Conclusion: The ESRβ gene holds potential as a biomarker for breast cancer prognosis

Amplification of Tumor Transcripts from Limited Quantity of Esophageal Squamous Cell Carcinoma Tissue Samples

To examine the template-switching technology accompanied by in vitro transcription (the Switch Mechanism At the 5ʹ end of Reverse Transcript) to amplify enough amount of mRNA as input for gene expression experiments. We amplified limited quantity of esophageal squamous cell carcinoma (ESCC) transcripts samples using generated ds cDNA as template and in vitro transcription (IVT) reaction. In addition, the quality and quantity of amplified mRNA were assessed by comparative real-time PCR of genes such as stem cell markers CD44, OCT4 and SNAIL as well as MAGE-A4 as a cancer-testis antigens, and XRCC5 as an underexpressed gene in ESCC.The results obtained from this study demonstrated that optimal amounts of mRNA are generated by template-switching and IVT reaction. Integrity and purity of all RNA samples were assessed. By using this approach, over 10 micrograms of amplified mRNA were generated from 100 ng of starting total RNA. The results of comparative real-time PCR of five genes with different levels of expression illustrated that the expression level of amplified sense RNA was almost similar when compared with non-amplified RNA. Our results clearly showed the usefulness of the T7-based IVT technique for amplification of limited quantity of input total RNA

The Role of Strogen Receptor β Expression and its Relationship with Tumor Characteristics in Patients with Breast Cancer

Background and purpose: Breast cancer is the 1st most common cancer in the female population and the third leading cause of cancer mortality in Ladies. Given the importance of early detection of cancer, it is necessary to find biomarkers related to this disease that are essential for the prognosis and early diagnosis of breast cancer. The estrogen receptor gene for beta, whose full name is Estrogen Receptor-β (ESRβ). Our main aim in this study was to investigate the expression changes of this gene in cancer tissue tumor samples compared to normal tissue samples to obtain a biomarker to predict the risk of cancer. Materials and methods: In this research, in 2023 samples were obtained from 30 patients diagnosed with breast cancer who underwent surgery at Omid Hospital in Mashhad. Both tumor tissue and adjacent healthy tissue were collected from each patient. The diagnosis of breast cancer was confirmed by a specialist doctor, and none of the patients had received any prior treatment such as chemotherapy or radiation therapy. Tissue samples were collected immediately after surgery and stored at -70°C. RNA extraction and synthesis were performed, followed by analysis of the expression of the ESR gene along with the GAPDH gene as an internal control, using real-time PCR. Results: In this study, 30 cancer patients were examined, with ages ranging from 32 to 62 years old and a mean age of 47.5 ± 9.38. Among them, one person had tumor grade 1, 16 had grade 2 tumors, and 8 had grade 3 tumors. Metastasis was observed in 11 patients (36.7%), while 19 patients (63.3%) had no metastasis. Regarding tumor size distribution, 5 patients (16.7%) had tumors smaller than 1 cm, and 25 patients (83.3%) had tumors sized between 1 and 2 cm. The study focused on changes in the expression of the ESRβ gene in both tumor and adjacent healthy tissues of breast cancer patients. It was found that the gene expression was higher in tumor samples compared to the adjacent healthy tissues. Using SPSS software, the analysis revealed a significant relationship between gene expression intensity and tumor grade across all three groups. Additionally, a significant difference was observed between patients with and without tumor metastasis. Conclusion: Overall, the findings of this study indicate a significant correlation between gene expression intensity and tumor grade across all three groups. Moreover, there is a significant distinction between patients with and without tumor metastasis. Notably, a considerable expression difference was observed between healthy and tumor samples. Hence, this gene could serve as a potential biomarker for cancer diagnosis

Expression analysis of CD44 isoforms S and V3, in patients with esophageal squamous cell carcinoma

Objective(s):  CD44 is a member of the cell adhesion molecules family. Naturally, CD44S, along with CD44V3 influence the cell motility, migration, and adhesion, while in tumor cells they lead to tumor invasion, progression, and metastasis. The purpose of this research is to evaluate the CD44S and CD44V3 expression in Esophageal Squamous Cell Carcinoma (ESCC) and to reveal their correlations with clinicopathological features of patients. Materials and Methods:Fresh tumoral and distant tumor-free esophageal tissues were obtained from 50 patients with ESCC. Using quantitative real-time PCR, the expression levels of CD44S and CD44V3 were quantified and compared in both groups of cells. The patients had not received any therapeutic interference, such as chemotherapy or radiation, prior to sampling. Results:Significant overexpression of CD44S and CD44V3 mRNA was observed in 13 (26.0%, P=0.03) and 11 (22.0%, P=0.007) tumor specimens, respectively. The expression of the genes were significantly correlated not only with each other (P=0.0001), but also with differentiation grade of tumor (P=0.033), stage of tumor progression (P=0.003), and depth of tumor invasion         (P=0.00). In addition, low level of CD44V3 mRNA expression was attended to be associated with tumor invasion. Conclusion:There is no correlation between CD44S expression with clinicopathological features of patients; however, simultaneous expression of these genes has an important effect on tumorigenesis

Antioxidant Effects of Statins by Modulating Nrf2 and Nrf2/HO-1 Signaling in Different Diseases

Statins are competitive inhibitors of hydroxymethylglutaryl-CoA (HMG-CoA) reductase and have been used to treat elevated low-density lipoprotein cholesterol (LDL-C) for almost four decades. Antioxidant and anti-inflammatory properties which are independent of the lipid-lowering effects of statins, i.e., their pleiotropic effects, might be beneficial in the prevention or treatment of many diseases. This review discusses the antioxidant effects of statins achieved by modulating the nuclear factor erythroid 2 related factor 2/ heme oxygenase-1 (Nrf2/HO-1) pathway in different organs and diseases. Nrf2 and other proteins involved in the Nrf2/HO-1 signaling pathway have a crucial role in cellular responses to oxidative stress, which is a risk factor for ASCVD. Statins can significantly increase the DNA-binding activity of Nrf2 and induce the expression of its target genes, such as HO-1 and glutathione peroxidase) GPx, (thus protecting the cells against oxidative stress. Antioxidant and anti-inflammatory properties of statins, which are independent of their lipid-lowering effects, could be partly explained by the modulation of the Nrf2/HO-1 pathway

Evaluation of Oxidative Stress Status in Familial Hypercholesterolemia

Background: Familial hypercholesterolemia (FH) is an autosomal dominant genetic disorder characterizied by elevated levels of circulating low-density lipoprotein cholesterol (LDL-C) which is an important source of substrates to be oxidized by different oxidative agents. Subsequently, the oxidized LDLs (oxLDLs) induce further oxidative reactions in FH patients, which contributes to the development of atherosclerosis and advanced cardiovascular events in these patients. This study aimed to investigate the association of oxidant/antioxidant markers with FH. Methods: This case-control study comprised 18 HoFH, 18 HeFH, and 20 healthy subjects. Oxidant/antioxidant markers including MDA, MPO, thiol, nitric oxide (NO), myeloperoxidase (MPO), glutathione peroxidase (GPx), SOD, and CAT were assessed by colorimetric methods. Prooxidant-antioxidant balance was also measured by pro-oxidant antioxidant balance (PAB) assay. Results: The levels of MDA (p < 0.001), MPO activity (p < 0.001), thiol (p < 0.001), NO (p < 0.01), and PAB (p < 0.001) were notably higher in HoFH group in comparison with healthy subjects. HeFH group also showed significantly higher levels of thiol (p < 0.001) and PAB (p < 0.001) when compared to healthy subjects. Elevated levels of MDA (p < 0.001) and PAB (p < 0.001) were also observed in HoFH relative to HeFH. No significant differences were found between the studied groups in the case of antioxidant enzyme activities. The results of binary logistic regression showed that PAB (OR: 0.979; p = 0.033), and MDA (OR: 0.996; p = 0.018) levels were inversely associated with HoFH, although, after adjustment for age and LDL-C levels, these associations were diminished. Conclusion: Several oxidant/antioxidant differences were found between FH patients and healthy individuals as well as between HoFH and HeFH patients. These differences might be strongly dependent on plasma LDL-C levels

The potential application of organoids in breast cancer research and treatment

Tumor heterogeneity is a major challenge for breast cancer researchers who have struggled to find effective treatments despite recent advances in oncology. Although the use of 2D cell culture methods in breast cancer research has been effective, it cannot model the heterogeneity of breast cancer as found within the body. The development of 3D culture of tumor cells and breast cancer organoids has provided a new approach in breast cancer research, allowing the identification of biomarkers, study of the interaction of tumor cells with the microenvironment, and for drug screening and discovery. In addition, the possibility of gene editing in organoids, especially using the CRISPR/Cas9 system, is convenient, and has allowed a more detailed study of tumor behavior in models closer to the physiological condition. The present review covers the application of organoids in breast cancer research. The recent use of gene-editing systems to provide insights into therapeutic approaches for breast cancer, is highlighted. The study of organoids and the possibility of gene manipulation may be a step towards the personalized treatment of breast cancer, which has so far remained unattainable due to the high heterogeneity of breast cancer