The lncRNA expression profile signature of leukemia stem cells is altered upon PI3K/mTOR inhibition: an <i>in vitro</i> and <i>in silico</i> study

Genetic and/or epigenetic alterations in hematopoietic stem cells (HSCs) contribute to leukemia stem cell (LSC) formation. We aimed to identify alterations in the lncRNA expression profile signature of LSCs upon inhibition of PI3K/Akt/mTOR signaling, which provides selective advantages to LSCs. We also aimed to elucidate the potential interaction networks and functions of differentially expressed lncRNAs (DELs). We suppressed PI3K/Akt/mTOR signaling in LSC and HSC cell-lines by specific PI3K/mTOR dual-inhibitor (VS-5584) and confirmed the inhibition by antibody-array. We defined DELs by qRT-PCR. Increased SRA, ZEB2-AS1, antiPeg11, DLX6-AS, SNHG4, and decreased H19, PCGEM1, CAR-Intergenic-10, L1PA16, IGF2AS, and SNHG5 levels (|log2fold-change|>5) were strictly associated with PI3K/Akt/mTOR pathway inhibition in LSC. We performed in silico analyses for DELs. ZEB2-AS1 was found to be specifically expressed in normal bone marrow and predominantly lower in leukemic cell-lines. Three sub-clusters were identified for DELs and they were associated with “abnormality of multiple cell lineages in the bone marrow.” DELs were most highly enriched for “glucuronidation” Reactome pathway and “ascorbate and aldarate metabolism” and “inositol phosphate metabolism” KEGG pathways. Transcription factors, MBD4, NANOG, PAX6, RELA, CEBPB, and CEBPA were predicted to be associated with the DEL profile. SRA was predicted to interact with CREB1, RARA, and PPARA. The possible DELs’ targets were predicted to form six ontological groups, be highly enriched for phosphoprotein, and be involved in “PPAR signaling pathway” and “ChREBP regulation by carbohydrates and cAMP.” These results will help to elucidate the roles of lncRNAs in the mechanisms that provide selective advantages to leukemia stem cells.</p

Effect of CCT137690 on long non-coding RNA expression profiles in MCF-7 and MDA-MB-231 cell lines

Gunduz, Cumhur/0000-0002-6593-3237WOS: 000512952000007PubMed: 31319040Long non-coding RNAs (lncRNAs) are involved in a range of biological processes, such as cellular differentiation, migration, apoptosis, invasion, proliferation, and transcriptional regulation. the aberrant expression of lncRNAs plays a significant role in several cancer types. Aurora kinases are increasingly expressed in various malignancies; accordingly, the inhibition of these enzymes may represent a novel approach for the treatment of various cancers. CCT137690, an Aurora kinase inhibitor, displays an anti-proliferative activity in human cancer cell lines. the aim of the present study was to investigate the anti-proliferative and cytotoxic effects of CCT137690 on estrogen receptor (ER)-positive human breast cancer cell line (MCF-7) and ER-negative human breast cancer cell line (MDA-MB-231). in addition, this study was targeted toward determining the changes induced in lncRNA expression levels following the initiation of Aurora kinase inhibitor treatment. the cytotoxic effects of CCT137690 were determined by means of the xCELLigence system. Furthermore, the anti-proliferative role of CCT137690 in breast cancer was investigated by checking the changes in lncRNA expression profiles using quantitative reverse-transcription polymerase chain reaction (qRT-PCR). the half-maximal inhibitory concentrations (IC50) of CCT137690 were determined as 4.5 mu M (MCF-7) and 7.27 mu M (MDA-MB-231). Several oncogenic lncRNAs (e.g., PRINS, HOXA1AS, and NCRMS) were downregulated in both ER-negative and ER-positive cell lines. on the other hand, tumor suppressor lncRNAs (e.g., DGCR5 and IGF2AS) were upregulated in the ER-positive cell line. After CCT137690 treatment, HOXA11AS and PCAT-14. lncRNAs were downregulated in the ER-positive cell lines. in addition, MER11C, SCA8, BC200, HOTAIR, PCAT-1, UCA1, SOX2OT, and HULC lncRNAs were downregulated in the ER-negative cell lines. the results of the present study indicated that Aurora kinase inhibitor CCT137690 could be a potential anti-cancer agent for breast cancer treatment

Nuclear imaging potential and in vitro photodynamic activity of symmetrical and asymmetrical zinc phthalocyanines

WOS: 000374698900005PubMed ID: 27059543Photodynamic therapy (PDT) is based on exposing a light-sensitive material that has been localized in target tissues with visible light. In the current study, symmetric Zn(II) octaoctadodecylphthalocyanine (1) and the asymmetrically substituted hydroxyhexyloxy derivative (2) were examined as a multifunctional agent for tumour nuclear imaging and for PDT potential. Zn(II)Pc 1 and Zn(II)Pc 2 were radiolabelled with I-131 using an iodogen method with high efficiency (93.5 +/- 3.5% and 93.0 +/- 2.8%, respectively) under the optimum conditions. Biodistribution study results showed that radiolabelled Zn(II)Pc 1 had a high uptake in the large intestine and unchanging uptake in the ovary. However, radiolabelled Zn(II)Pc 2 uptake was statically significant in the large intestine, pancreas, ovary and lung. For the PDT studies, EMT6/P (mouse mammary cell line) and HeLa (cervical adenocarcinoma cell line) with Zn(II)Pc 1 and Zn(II)Pc 2 were exposed to red light (650nm) at 10-30J/cm(2). Zn(II)Pc 1 and Zn(II)Pc 2 had a good PDT efficacy in the EMT6/P cell line. In conclusion, radiolabelled Zn(II)Pc 1 might be a promising imaging agent for pancreas, ovary and colon tumours. However, the radiolabelled Zn(II)Pc 2 might be a promising nuclear imaging and PDT agent for colon, lung, pancreas and ovary tumours

Design, synthesis, cytotoxic activity, and apoptosis inducing effects of 4-and N-substituted benzoyltaurinamide derivatives

akgul, ozlem/0000-0002-0011-7888; Birim, Dervis/0000-0002-5094-9949; Gunduz, Cumhur/0000-0002-6593-3237; Erdogan, Mumin/0000-0003-0048-444XWOS:000599807000020PubMed: 33488262In this study, a group of 4-substituted benzoyltaurinamide derivatives were designed, synthesized, and investigated for their anticancer activity against three cancer cell lines and one nontumorigenic cell line by MTT assay. Among the final compounds, methoxyphenyl derivatives 14, 15, 16 were found to be effective against all the tested cancerous cell lines with promising selectivity. The most active compounds were further evaluated to determine the molecular mechanism of their anticancer activity by using western blot assay and the Annexin V-FITC/PI test. Compound 14 (in SH-SY5Y and MDA-MB-231 cell lines) and 15 (in SH-SY5Y cell line) were found to induce intrinsic apoptotic pathway by upregulating BAX, caspase-3, and caspase-9, while downregulating Bcl-2 and Bcl-xL expression levels. According to mechanistic studies, compounds displayed their anticancer activity via three different mechanisms: a. caspase-dependent, b. caspase-independent, and c. caspase-dependent pathway that excluded caspase-9 activation. As a result, this study provides interesting data which can be used to design new taurine-based anticancer derivatives.Scientific and Technological Research Council of Turkey [TUBITAK]Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [SBAG-118S667]This study was supported by research grants from The Scientific and Technological Research Council of Turkey [TUBITAK; Project number: SBAG-118S667]

Analysis of dysregulated long non-coding RNA expressions in glioblastoma cells

WOS: 000381539800015PubMed ID: 27306825Long non coding RNAs (IcRNAs) are associated with various biological roles such as embryogenesis, stem cell biology, cellular development and present specific tissue expression profiles. Aberrant expression of IncRNAs are thought to play a critical role in the progression and development of various cancer types, including gliomas. Glioblastomas (GBM) are common and malignant primary brain tumours. Brain cancer stem cells (BCSC) are isolated from both low and high-grade tumours in adults and children, by cell fraction which express neuronal stem cell surface marker CD133. The purpose of this study was to investigate the expression profiles of IncRNAs in brain tumour cells and determine its potential biological function. For this purpose, U118MG-U87MG; GBM stem cell series were used. Human parental brain cancer cells were included as the control group; the expressions of disease related human IncRNA profiles were studied by LightCycler 480 real-time PCR. Expression profiles of 83 lncRNA genes were analyzed for a significant dysregulation, compared to the control cells. Among IncRNAs, 51 IncRNA genes down-regulated, while 8 IncRNA genes were up-regulated. PCAT-1 (2.36), MEG3 (5.34), HOTAIR (-2.48) lncRNAs showed low expression in glioblastoma compared to the human (parental) brain cancer stem cells, indicating their role as tumour suppressor genes on gliomas. As a result, significant changes for anti-cancer gene expressions were detected with disease-related human IncRNA array plates. Identification of novel target genes may lead to promising developments in human brain cancer treatment. (C) 2016 Elsevier B.V. All rights reserved.Research Foundation of Ege UniversityEge University [2.101.2014.0014]This study was supported by the Research Foundation of Ege University (Grant Number 2.101.2014.0014)

Evaluation of nuclear imaging potential and photodynamic therapy efficacy of symmetrical and asymmetrical zinc phthalocyanines

WOS: 000378453600018Photodynamic therapy (PDT) is a medical treatment for the removal of target tissues involving the delivery of a photosensitizer agent followed by irradiation with visible light. In the present study, symmetric Zn(II)Pc 1 and asymmetrically substituted Zn(II)Pc 2 were synthesized and examined multifunctional agents for tumor nuclear imaging, and PDT potential. The Zn(II)Pc 1 and Zn(II)Pc 2 were radiolabeled with I-131 with high efficiency (93.4 +/- 1.6% and 91.4 +/- 1.6%, respectively). The results of the biodistribution study showed that radiolabeled Zn(II)Pc 1 had high uptake on lung, large intestine, ovary and pancreas. However, the uptake of radiolabeled Zn(II)Pc 2 was statically significant in pancreas and intestine. In PDT studies, EMT6/P (mouse mammary cell) and HeLa (cervical adenocarcinoma cell) with Zn(II)Pc 1 and Zn(II)Pc 2 were exposed to red light at the doses of 10-30 J/cm(2). Although PDT activity of Zn(II)Pc 2 in HeLa cell line was determined, Zn(II)Pc 1 showed no phototoxic effect in both cell lines. In conclusion, radiolabeled Zn(II)Pc 1 might be a promising imaging agent for the lung, the ovary pancreas, and the colon tumors. However, radiolabeled Zn(II)Pc 2 might be a promising nuclear imaging agent for the colon and the pancreas tumors and promising PDT agent for cervical tumors. (C) 2016 Elsevier B.V. All rights reserved.Scientific and Technological Research Council of Turkey, TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [112T565]The authors gratefully acknowledge the financial support by the Scientific and Technological Research Council of Turkey, TUBITAK (Grant no: 112T565)

The Evaluation of Effect of Aurora Kinase Inhibitor CCT137690 in Melanoma and Melanoma Cancer Stem Cell

Background: Dysregulation of the cell cycle is one of the main causes of melanomagenesis. Genomewide studies showed that the expression of Aurora-A and-B significantly has been upregulated in melanoma. However, there is no FDA approved drug targeting aurora kinases in the treatment of melanoma. In addition, the development of resistance to chemotherapeutic agents in the treatment of melanoma and, as a result, the relapse due to heterogeneous cell groups in patients is a second phenomenon that causes treatment failure. Therefore, there is an urgent need for therapeutic alternatives targeting both melanoma and melanoma cancer stem cells (MCSCs) in treatments. At this stage, cell cycle regulators become promising targets. Objective: In this study, we aimed to identify the effects of Aurora kinase inhibitor CCT137690 on the cytotoxicity, apoptosis, cell cycle, migration, and colony formation and expression changes of genes related to proliferation, cell death and cell cycle in melanoma and melanoma cancer stem cell. In addition, we investigated the apoptotic and cytostatic effects of CCT137690 in normal fibroblast cells. Methods: We evaluated the cytotoxic effect of CCT137690 in MCSCs, NM2C5 referring as melanoma model cells and WI-38 cells by using the WST-1 test. The effect of CCT137690 on apoptosis was detected via Annexin V and JC-1 method; on cell cycle progression by cell cycle test; on gene expression by using RT-PCR, on migration activity by wound healing assay and clonal growth by clonogenic assay in NM2C5 cells and MCSCs. The effects of CCT137690 in WI-38, referring as healthy fibroblast cell, were assessed through Annexin V and cell cycle method. Results: CCT137690 was determined to have a cytotoxic and apoptotic effect in MCSCs and melanoma. It caused polyploidy and cell cycle arrest at the G2/M phase in MCSCs and melanoma cells. The significant decrease in the expression of MMP2, MMP7, MMP10, CCNB1, IRAK1, PLK2 genes, and the increase in the expression of PTEN, CASP7, p53 genes were detected. Conclusion: Aurora kinases inhibitor CCT137690 displays promising anticancer activity in melanoma and especially melanoma cancer stem cells. The effect of CCT137690 on melanoma and MCSC may provide a new approach to treatment protocols