Genistein-induced mir-23b expression inhibits the growth of breast cancer cells

Aim of the study: Genistein, an isoflavonoid, plays roles in the inhibition of protein tyrosine kinase phosphorylation, induction of apoptosis, and cell differentiation in breast cancer. This study aims to induce cellular stress by exposing genistein to determine alterations of miRNA expression profiles in MCF-7 cells. Material and methods: XTT assay and trypan blue dye exclusion assays were performed to examine the cytotoxic effects of genistein treatment. Expressions of miRNAs were quantified using Real-Time Online RT-PCR. Results: The IC50 dose of genistein was 175 μM in MCF-7 cell, line and the cytotoxic effect of genistein was detected after 48 hours. miR-23b was found to be up-regulated 56.69 fold following the treatment of genistein. It was found that miR-23b was up-regulated for MCF-7 breast cancer cells after genistein treatment. Conclusions: Up-regulated ex-expression of miR-23b might be a putative biomarker for use in the therapy of breast cancer patients. miR-23b up-regulation might be important in terms of response to genistein. © 2015, Termedia Publishing House Ltd. All rights reserved

Effects of quercetin induced cell death on a novel gene "URG4/URGCP" expression in leukemia cells

The present study aimed to investigate anti-proliferative and apoptotic effects of quercetin on human leukemia cells and effects of quercetin-induced cell death on a novel gene Up-regulated gene 4/upregulator of cell proliferation (URG4/URGCP), in leukemia cells. URG4/URGCP expression is determined by using RT-PCR. IC 50 of quercetin was determined as 25 microM in CCRF-CEM, HL-60 and K562 cells. In IC 50 dose group, URG4/URGCP expression was decreased 99% in HL-60 cells, 90% in CCRF-CEM cells, and 52% (24 hour) - 99% (72 hour) in K-562 cells. URG4/URGCP may play important roles in the development of leukemia, and might be a useful molecular marker for predicting the prognosis of leukemia via quercetin treatment. © 2012 Dodurga Y, et al

Downregulation of miR-195 via cyclosporin a in human glioblastoma cells

PubMed ID: 26537083Purpose: Cyclosporin A (CsA) is a potent immunosuppressive agent. MicroRNAs (miRs) which post-transcriptionally regulate gene expression are non-coding RNAs. The aim of this study was to investigate the effects of CsA on 88 miRs expression changes in glioma cells (U-87 MG). Methods: CsA was used in U-87 MG glioma cells in doses of 10, 30 and 60 uM. Cytotoxic assays and determination of IC50 dose of CsA were performed. Relative quantification of 88 miRs was performed by real time RT-PCR. The fold changes of miRs determined and alterations in the miR expressions were compared with CsA-treated and CsA-free U-87 MG glioma cells. Results: In U-87 MG cells treated with CsA, the ICso dose was 10 µM. Seventeen of 88 human miRs were downregulated compared to the untreated control group by using miRs array. It was found that the expression levels of several miRs, in particular miR-195, was significantly decreased in CsA-treated U-87 MG cells. Conclusion: This study revealed a significant role of miR-195 in the molecular pathology of glioma cells which can also implicate potential application of miR-195 in cancer therapy. Rather than downregulation of miR-195 alone to exhibit cytotoxicity, treatment with CsA could be more effective especially on temozolomide-resistant cells

Effect of resveratrol on apoptosis and MDM2, RUNX3, RB gene expressions in human acute myeloid leukemia cells by transfection of MATRA-mediated miR-150 [MATRA aracılı miR-150’nin transfeksiyonu ile insan akut myeloid lösemi hücrelerinde resveratrol’ün MDM2,RUNX3,RB gen ekspresyonları ve apoptoz üzerine etkisi]

Acute myeloid leukemia (AML) is a heterogeneous group of hematopoietic malignancies. Magnet assisted transfection (MATRA) is one of the most effective non-viral transfection methods. We aimed to evaluate the apoptotic effect of resveratrol (RES), MDM2, RUNX3, RB gene expression exchanges and MicroRNA-150 (miR-150) transfected with MATRA to HL-60 cells. MATRA was used as non-viral vector carrier for miR-150 transfection. IC50 dose of RES was determineted by Fahri et al. Resveratrol and miR-150 transfected cells were performed apoptosis and MDM2, RUNX3, RB gene expression assays in Human promyelocytic leukemia (HL-60) cells. IC50 dose of RES was used as 5 µM. In HL-60 cells, it was found that miR-150, miR150-Resveratrol combination, resveratrol alone induces apoptosis by 6.48, 6.93 and 4.54 fold, respectively, compared to the control cells. Compared to control cells, MDM2, RUNX3, RB, gene expressions decreased (miR-150) 1.7, 1.3 and 1.4 fold, (miR-150-resveratrol) MDM2 expression increased 2.73 fold, RUNX3 and RB expression decreased 6.1, 1.07 fold, respectively. Combination with resveratrol, MDM2, RB decreased 2.9 and 1.4 fold, respectively. Non-viral miR-150 transfection may be effective in leukemia cells, induction of apoptotic effects and gene expression changes, following treatment with resveratrol and miR-150-resveratrol combinations. © 2018, UHOD - Uluslararasi Hematoloji Onkoloji Dergisi. All rights reserved

The emphasis of tumor suppressor genes and oncogenes in diagnosis and prognosis of anaplastic brain tumors [Anaplastik beyin tümörlerinin tani{dotless} ve prognozunda tümör süpressör genlerin ve onkogenlerin önemi]

The aim of the study is to the determine the profiles of tumor suppressor genes and oncogenes which cause brain tumor, establishing the association between the prognosis of cancer and the quantitation of genetic and epigenetic changes, and bringing a molecular approach to definite diagnosis. For this purpose, explant cell cultures are performed from the anaplastic brain tumor tissues of the cases. The expression analysis of the tumor suppressor genes (p53, RB1, PTEN, MGMT, RUNX3, DMBT1, PIKE) and oncogenes (EGFR, PIK3CA, MDM2, Olig2, GSTT1, COX-2 and hTERT) were determined by comparing the expression of GAPDH housekeeping gene using real-time online RT-PCR. The promoter regions of all the tumor suppressor genes' hypermethylation and also methylated and unmethylated copy numbers were determined with Q-PCR by using methylation specific primer and probes and the quantitation was carried out by comparing with each other. A significant difference was determined among the oncogenes; EGFR and hTERT gene expressions in patient tumor group. hTERT gene expression showed a significant difference with tumor grades. DMBT1 gene expression showed a significant difference with tumor grades. A prominent decrease was found in the aberration of tumor suppressor gene copy number in the glioma group. Gene copy number and gene expression of GSTT1 gene showed a significant correlation. RB1 and MGMT promoter methylation showed a significant difference in tumor patient group. Over expression of PIK3CA, EGFR and COX-2 among oncogenes and loss of copy number of PTEN, RB1 and RUNX3 among tumor suppressor genes found associated with short survival

Expression profiling of RE1-silencing transcription factor (REST), REST corepressor 1 (RC0R1), and Synapsin 1 (SYN1) genes in human gliomas

PubMed ID: 27685921Purpose: The repressor element 1 (RE-1) silencing transcription f actor (REST) is a transcription f actor which represses the expression of neuronal differentiation-related genes including SYN1 gene. CoREST, encoded by RCOR1 gene, binds to the REST protein for remodeling of chromatin structure. Although there is a relation among REST, RCOR1, and SYN1 genes, the role of these genes in glioma tumors is still unclear. In this study, expressions of REST, RC0R1, and SYN1 genes were detected in primary cultures derived from tumor samples of diffuse astrocytoma (DA), anaplastic oligodendroglioma (AO), and glioblastoma multiforme (GBM) cases. Methods: Expression profiles were analysed by RT-qPCR and the copy number variations were examined with qPCR in primary cultures. ChIP assay was performed to show binding characteristics of REST and CoREST proteins on promoter region of SYN1 gene. Results: Means of relative expression for REST were as follows: 0.7898, 0.7606, and 0.7318 in DA, AO, and GBM groups, respectively. For RCOR1, expression means in DA, AO, and GBM groups were 0.7203, 0.7334, and 0.7230, respectively. SYN1 expression means were as follows: 0.3936, 0.3192, and 0.3197 in DA, AO, and GBM groups, respectively. Neither gain nor loss of copy numbers were detected for REST and RC0R1 genes in all groups. Copy loss for SYN1 was detected in primary culture of a DA case. REST and CoREST presented positive precipitation pattern on promoter region of SYN1 gene. Conclusions: Expressions of REST and RC0R1 genes may downregulate SYN1 expression in gliomas. Low expression pattern of SYNl may maintain cancer stem-like phenotype which contributes to development of gliomas