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In vivo gene silencing

M Joaquina Delás, Benjamin T Jackson, Tatjana Kovacevic, Silvia Vangelisti, Ester Munera Maravilla, Sophia A Wild, Eva Maria Stork, Nicolas Erard, Simon R V Knott, Gregory J Hannon
Long non-coding RNAs (lncRNAs) show patterns of tissue- and cell type-specific expression that are very similar to those of protein coding genes and consequently have the potential to control stem and progenitor cell fate decisions along a differentiation trajectory. To understand the roles that lncRNAs may play in hematopoiesis, we selected a subset of mouse lncRNAs with potentially relevant expression patterns and refined our candidate list using evidence of conserved expression in human blood lineages. For each candidate, we assessed its possible role in hematopoietic differentiation in vivo using competitive transplantation...
April 16, 2019: Cell Reports
Qin Xiang, Dishu Zhou, Xiaoqian He, Jiangxia Fan, Jun Tang, Zhu Qiu, Yan Zhang, Jingfu Qiu, Yongzhu Xu, Guoqi Lai
BACKGROUND AND AIM: The high mortality and poor prognosis of hepatocellular carcinoma (HCC) have raised the public attention. Gene therapy is considered as a promising treatment option for cancer; thus, finding a new therapeutic target for hepatocellular carcinoma is urgently needed. GATA4 is a tumor suppressor gene in multiple cancers, but its role in HCC is unclear. In this study, we explored the function of GATA4 in HCC. METHODS: RT-PCR (Reverse Transcription-Polymerase chain reaction) and qPCR (quantitative PCR) were used to detect the mRNA expression of GATA4 in HCC cells and tissues...
April 17, 2019: Journal of Gastroenterology and Hepatology
Hua Tang, Xiaopeng Xu, Weirong Xiao, Yangying Liao, Xiao Xiao, Lan Li, Ke Li, Xiaomin Jia, Hao Feng
Melanoma is considered as an aggressive neoplastic transformation and featured with high metastatic potential. Although some studies have provided targets for novel therapeutic interventions, clinical development of targeted drugs for melanoma still remains obscure. Therefore, this study aims to identify the role of microRNA-27a (miR-27a) in autophagy and apoptosis of melanoma cells in regulating spleen tyrosine kinase (SYK)-mediated the mammalian target of rapamycin (mTOR) signaling pathway. A microarray-based analysis was made to screen differentially expressed genes and predict target miRNA...
April 17, 2019: Journal of Cellular Biochemistry
Yoshiyuki Hattori, Satono Shimizu, Kei-Ichi Ozaki, Hiraku Onishi
In this study, we examined the effect of cationic lipid type in folate (FA)-polyethylene glycol (PEG)-modified cationic liposomes on gene-silencing effects in tumor cells using cationic liposomes/siRNA complexes (siRNA lipoplexes). We used three types of cationic cholesterol derivatives, cholesteryl (3-((2-hydroxyethyl)amino)propyl)carbamate hydroiodide (HAPC-Chol), N -(2-(2-hydroxyethylamino)ethyl)cholesteryl-3-carboxamide (OH-Chol), and cholesteryl (2-((2-hydroxyethyl)amino)ethyl)carbamate (OH-C-Chol), and we prepared three types of FA-PEG-modified siRNA lipoplexes...
April 15, 2019: Pharmaceutics
You-Cheng Hseu, Yi-Geng Ho, Dony Chacko Mathew, Hung-Rong Yen, Xuan-Zao Chen, Hsin-Ling Yang
Coenzyme CoQ10 (CoQ10), a ubiquinone compound, has been reported to inhibit tyrosinase activity and melanin production in melanoma B16F10 cells. However, the molecular mechanism underlying this inhibitory effect is poorly understood. In this paper we aimed to investigate the molecular mechanisms involved in the anti-melanogenic activity of CoQ10 (1-2 μM) in UVA (5 J/cm2 )-irradiated keratinocyte HaCaT cells and α-MSH stimulated B16-F10 cells. It was observed that CoQ10 suppressed p53/POMC, α-MSH production as well as inhibited ROS generation in UVA-irradiated keratinocyte HaCaT cells...
April 13, 2019: Biochemical Pharmacology
Harika Sabbineni, Arti Verma, Sandeep Artham, Daniel Anderson, Oge Amaka, Fang Liu, Subhadra P Narayanan, Payaningal R Somanath
Endothelial to mesenchymal transition (EndMT), where endothelial cells acquire mesenchymal characteristics has been implicated in several cardiopulmonary, vascular and fibrotic diseases. The most commonly studied molecular mechanisms involved in EndMT include TGFβ, Notch, interleukin, and interferon-γ signaling. As of today, the contributions of Akt1, an important mediator of TGFβ signaling and a key regulator of endothelial barrier function to EndMT remains unclear. By using the ShRNA based gene silencing approach and endothelial-specific inducible Akt1 knockdown (ECKOAkt1 ) mice, we studied the role of Akt1 in EndMT in vitro and pathological vascular remodeling in vivo...
April 13, 2019: Biochemical Pharmacology
Yong Tang, Yiwen Xuan, Guibin Qiao, Zhu'an Ou, Zhe He, Qihang Zhu, Ming Liao, Guilin Yin
Background: Mouse double minute 2 (MDM2) contributes to cancer metastasis and epithelial-mesenchymal transition (EMT). This study aimed to investigate small mothers against decapentaplegic (Smad) signaling in MDM2-mediated EMT in lung adenocarcinoma (LAC). Materials and methods: Expression patterns of MDM2 in LAC tissues, adjacent tissues, and cell lines (BEAS-2B, PC9, H1975, and A549) were detected. We then overexpressed MDM2 in PC9 cells and knocked it down in H1975 cells...
2019: OncoTargets and Therapy
Hongtao Zhu, Xiaoling Gan, Xingwei Jiang, Shuai Diao, Huan Wu, Jianguo Hu
BACKGROUND: ALKBH5 regulated the malignant behavior of breast cancer and glioblastoma. However, the expression and function of ALKBH5 in epithelial ovarian cancer have not yet been determined. In the present study, we investigated the expression and function of ALKBH5 in epithelial ovarian cancer with respect to its potential role in the tumorigenesis of the disease as well as an early diagnostic marker. METHODS: Immunohistochemistry and western blot were used to detect protein expression...
April 15, 2019: Journal of Experimental & Clinical Cancer Research: CR
Changmai Chen, Zhongyu Wang, Jinhao Zhang, Xinli Fan, Luzheng Xu, XinJing Tang
RNA interference (RNAi)-based gene therapy is a precision therapeutic approach for highly efficient sequence-specific gene silencing in vivo or in vitro. Caged RNAs featuring dextran conjugation of antisense and sense RNA strands using photolabile linker were rationally designed and self-assembled to form caged siRNA nanoparticles (Dex-p-siRNA) for photoregulation of target gene expression. The dextran-conjugated caged siRNA nanoparticles showed significant se-rum nuclease-resistance due to the formation of dextran-siRNA nanoparticles...
April 16, 2019: Bioconjugate Chemistry
Denghui Zhu, Rong Huang, Peipei Fu, Liangming Chen, Lifei Luo, Pengfei Chu, Libo He, Yongming Li, Lanjie Liao, Zuoyan Zhu, Yaping Wang
Basic leucine zipper transcription factor ATF-like (BATF)-3, belonging to activator protein 1 (AP-1) superfamily transcription factors, is essential for homeostatic development of CD8α⁺ classical dendritic cells activating CD8 T-cell responses to intracellular pathogens. In this study, the characteristics and cDNA cloning of the CiBATF3 molecule were described in grass carp ( Ctenopharyngodon idella ). CiBATF3 had abundant expression in immune-related organizations, including liver, spleen and gill, and grass carp reovirus (GCRV) infection had significantly changed its expression level...
April 4, 2019: International Journal of Molecular Sciences
Tian Tang, Guang Shan
Cancer stem cells (CSCs) have been recognized as the significant cause of tumor recurrence. Long noncoding RNAs (lncRNAs) are involved in various cancers, including human laryngeal cancer. So far the correlation between lncRNA DiGeorge syndrome critical region gene 5 (DGCR5) and CSC-like properties in human laryngeal cancer remains barely known. In our current study, two human larynx squamous carcinoma cell lines (Hep-2 and Hep-2R) with different radio sensitivities were cultured. Interestingly, CSC-like phenotypes were much more enriched in Hep-2R cells...
April 13, 2019: Journal of Cellular Physiology
Lisha Ai, Shidai Mu, Chunyan Sun, Fengjuan Fan, Han Yan, You Qin, Guohui Cui, Yadan Wang, Tao Guo, Heng Mei, Huafang Wang, Yu Hu
BACKGROUND: Myeloid-derived suppressor cells (MDSCs) and cancer stem cells (CSCs) are two important cellular components in the tumor microenvironment, which may modify the cancer phenotype and affect patient survival. However, the crosstalk between MDSCs and multiple myeloma stem cells (MMSCs) are relatively poorly understood. METHODS: The frequencies of granulocytic-MDSCs (G-MDSCs) in MM patients were detected by flow cytometry and their association with the disease stage and patient survival were analyzed...
April 13, 2019: Molecular Cancer
Aijaz A John, Ravi Prakash, Divya Singh
MicroRNAs (miRNAs) have appeared as critical controllers of gene expression at post-transcriptional level either by degrading RNA transcripts or repressing translation. It is evident from the ever growing scientific literature that microRNAs play a significant role in osteoblast commitment and differentiation. Here, we report that over-expression of miR-487b-3p leads to inhibition of osteoblastic differentiation. Using in silico approaches, Nrarp was found to be the direct target of miR-487b-3p which was further validated by luciferase 3' UTR reporter assay...
April 1, 2019: Journal of Endocrinology
Sue-Hong Wang, Shu-Yun Tung, Kuan-Chung Su, Hsiao-Hsuian Shen, Jia-Yang Hong, Ming-Shiun Tsai, Gunn-Guang Liou
To study the epigenetic gene silencing, yeast is an excellent model organism. Sir proteins are required for the formation of silent heterochromatin. Sir2 couples histone deacetylation and NAD hydrolysis to generate an endogenous epigenetic metabolic small molecule, O-acetyl-ADP-ribose (AAR). AAR is involved in the conformational change of SIR complexes, modulates the formation of SIR-nucleosome pre-heterochromatin and contribute to the spreading of SIR complexes along the chromatin fiber to form extended silent heterochromatin regions...
April 11, 2019: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Wei Wang, Jiayi Wu, Xiaochun Fei, Weiguo Chen, Yafen Li, Kunwei Shen, Li Zhu
Chromodomain helicase/ATPase DNA-binding protein 1-like (CHD1L) gene is a novel oncogene amplified in many solid tumors. We investigated its role in breast cancer. CHD1L was over-expressed in 49.1% (57/116) breast cancer patients. Overexpression of CHD1L was significantly associated with younger age at diagnosis ( P = 0.016), lymph node involvement ( P = 0.040), higher tumor grade ( P = 0.027), higher proliferation rate ( P = 0.007) and shorter disease-free survival rate (77.2% vs. 91.5%, P = 0.037). A cDNA microarray analysis identified MDM2 as an important downstream target of CHD1L...
2019: American Journal of Translational Research
Caixia An, Hailong Li, Xueyan Zhang, Jing Wang, Yi Qiang, Xinhua Ye, Qiang Li, Quanlin Guan, Yongning Zhou
Emerging studies have reported that coatomer protein complex subunit β2 (COPB2) is overexpressed in several types of malignant tumor; however, to the best of our knowledge, no studies regarding COPB2 in gastric cancer have been published thus far. Therefore, the present study aimed to determine the significance and function of COPB2 in gastric cancer. COPB2 expression in gastric cancer cell lines was measured using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. In addition, lentivirus‑short hairpin RNA (shRNA) COPB2 (Lv‑shCOPB2) was generated and used to infect BGC‑823 cells to analyze the effects of COPB2 on the cancerous phenotype...
February 18, 2019: International Journal of Oncology
Hans-Hermann Wessels, Svetlana Lebedeva, Antje Hirsekorn, Ricardo Wurmus, Altuna Akalin, Neelanjan Mukherjee, Uwe Ohler
MicroRNAs (miRNAs) are key mediators of post-transcriptional gene expression silencing. So far, no comprehensive experimental annotation of functional miRNA target sites exists in Drosophila. Here, we generated a transcriptome-wide in vivo map of miRNA-mRNA interactions in Drosophila melanogaster, making use of single nucleotide resolution in Argonaute1 (AGO1) crosslinking and immunoprecipitation (CLIP) data. Absolute quantification of cellular miRNA levels presents the miRNA pool in Drosophila cell lines to be more diverse than previously reported...
April 9, 2019: Nature Communications
Huaner Ni, Weifeng Li, Ying Zhuge, Shuang Xu, Yue Wang, Yang Chen, Gu Shen, Fang Wang
BACKGROUND: Circular RNAs (circRNAs) are emerging as powerful regulators of cardiac development and disease. Nevertheless, detailed studies describing circRNA-mediated regulation of cardiac fibroblasts (CFs) biology and their role in cardiac fibrosis remain limited. METHODS: PCR and Sanger sequencing were performed to identify the expression of circHIPK3 in CFs. Edu corporation assays, Transwell migration assays, and immunofluorescence staining assays were conducted to detect the function of circHIPK3 in CFs in vitro...
April 2, 2019: International Journal of Cardiology
Longwei Xu, Yuanyuan Su, Yichao Zhao, Xincheng Sheng, Renyang Tong, Xiaoying Ying, Lingchen Gao, Qingqi Ji, Yu Gao, Yang Yan, Ancai Yuan, Fujian Wu, Feng Lan, Jun Pu
Exercise-induced physiological hypertrophy provides protection against cardiovascular disease, whereas disease-induced pathological hypertrophy leads to heart failure. Emerging evidence suggests pleiotropic roles of melatonin in cardiac disease; however, the effects of melatonin on physiological versus pathological cardiac hypertrophy remain unknown. Using swimming-induced physiological hypertrophy and pressure-overload-induced pathological hypertrophy models, we found that melatonin treatment significantly improved pathological hypertrophic responses accompanied by alleviated oxidative stress in myocardium but did not affect physiological cardiac hypertrophy and oxidative stress levels...
April 8, 2019: Journal of Pineal Research
Ying-Yi Luan, Lei Zhang, Fu-Jun Zhu, Ning Dong, Jiang-Yang Lu, Yong-Ming Yao
Dendritic cell (DC) dysfunction plays a pivotal role in sepsis-induced immunosuppression. TIPE1, a new member of the TNFAIP8 (tumor necrosis factor-α-induced protein 8) family, may be related to cell death. The aim of present study was to elucidate the effect of TIPE1 on immune function of DCs and its regulatory mechanism via PD-L1/PD-1 signaling in mice. Sepsis was induced in adult C57BL/6 male mice via cecal ligation and puncture (CLP). In vitro, we found that expressions of CD80, CD86, MHC-II in DCs, and cytokines including TNF-α as well as IL-12p40 levels were elevated, similarly, T-cell proliferation and differentiation were promoted when TIPE1 gene was silenced...
April 8, 2019: Journal of Infectious Diseases
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