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dual luciferase reporter gene assay

Yanbo Yu, Mengye Zhu, Yong Zhao, Mu Xu, Mizhen Qiu
OBJECTIVE: The aim of this study was to investigate the mechanisms of TUSC7/miR-449a/PPAR-γ axis on the inflammation induced by microglia activation. METHODS: A compressive spinal cord injury (SCI) model was established. The expressions of TUSC7, miR-449a PPAR-γ, TNF-α and IL-1β in spinal cord tissues of SCI rats and HAPI cells were determined. The interaction of TUSC7 and miR-449a was tested by RIP and RNA pull-down assays. The regulatory relationship between miR-449a and PPAR-γ was tested by dual luciferase reporter gene assay...
September 5, 2018: Biochemical and Biophysical Research Communications
Binghua Zhou, Lijun Li, Yajing Li, Huimin Sun, Changchun Zeng
The long non-coding RNA small nucleolar RNA host gene 12 (SNHG12) overexpression was found in various tumors and the dysregulated expression of SNHG12 contributed to multidrug resistance in non-small cell lung cancer. However, the role of SNHG12 in doxorubicin resistance of osteosarcoma is still unclear. In present study, we explored the function and underlying mechanism of SNHG12 on doxorubicin resistance in osteosarcoma. High expression of SNHG12 was associated with doxorubicin resistance and a poor overall survival in osteosarcoma...
July 11, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Yang Zhang, Zhen Su, Hai-Lin Liu, Lin Li, Meng Wei, Dong-Jian Ge, Zhi-Jie Zhang
MicroRNA are emerging as significant regulators of neuropathic pain progression. In addition, neuroinflammation contributes a lot to neuropathic pain. miR-26a-5p has been identified as an inflammation-associated miRNA in multiple pathological processes. However, little is known about the biological role of miR-26a-5p in neuroinflammation and neuropathic pain development. Therefore, we aimed to investigate the function of miR-26a-5p in neuropathic pain by establishing a rat model using chronic sciatic nerve injury (CCI)...
August 14, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Chao-Zhi Tang, Jun-Tang Yang, Qing-Hui Liu, Ya-Ru Wang, Wen-Sheng Wang
Depression represents a condition characterized by cognitive deficits and neural dysfunction and has recently been correlated with microRNAs (miRs) and their respective target genes. The present study was conducted with the goal of investigating the expression of miR-192-5p and its target gene fibulin (Fbln)-2 in an attempt to evaluate their roles in the occurrence and progression of cognitive impairment and neural function in mice with chronic unpredictable mild stress (CUMS)-induced depression through regulation of the TGF-β1 signal transduction pathway...
August 17, 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Yun Zhang, Lingyun Shao
The abnormal expression of miRNAs (miRs) has previously been reported in respiratory syncytial virus (RSV) disease. However, to the best of our knowledge, the expression of miR-140-5p in patients with an RSV infection has never been explored. Reverse transcription-quantitative polymerase chain reaction was performed to analyze the level of miR-140-5p in the blood and nasopharyngeal airway samples. ELISAs were performed to determine the levels of tumor necrosis factor α, interleukin (IL)-1β, IL-6 and IL-8...
August 2018: Experimental and Therapeutic Medicine
Peng Li, Mingqiang Dong, Zhigang Wang
Previous studies demonstrated dysregulation of different microRNAs in thyroid cancer. Tetraspanins (TSPANs) are cell surface proteins with critical roles in many cellular processes, and implications in tumor development. Here we investigated the role of miR-369-3p in papillary thyroid cancer (PTC) and its association with TSPAN13. miR-369-3p and the TSPAN13 gene expression profiles of 513 thyroid cancer and 59 normal thyroid tissues were downloaded from the Cancer Genome Atlas database. Thyroid cancer tissues were classified according to the histological type, grouped based on low and high median miR-369-3p and TSPAN13 expression, and analyzed in relation to overall survival (OS) of patients...
August 2, 2018: Bosnian Journal of Basic Medical Sciences
Xiangqian Zhang, Min Zhang, Guifeng Wang, Ye Tian, Xiaolong He
It has previously been demonstrated that miRNA (miR)‑647 exhibits an important role in various cancers, and is aberrantly expressed in gastric cancer (GC). However, the exact role of miR‑647 in GC still remains unclear. The present study aimed to investigate the functional significance of miR‑647 and its target gene in GC. TargetScan and Miranda databases were used to predict the putative targets, and the prediction was validated by Dual‑luciferase Reporter Assays. To investigate whether miR‑647 affects GC cell behavior, a stable miR‑647‑overexpression/low‑expression cell line was generated by transfection with miR‑647 mimic/inhibitor...
August 6, 2018: Molecular Medicine Reports
Wei Lu, Xiaohua Tao, Yibin Fan, Yi Tang, Xin Xu, Shasha Fan, Youming Huang, Yong Yu, Dan Luo
Objectives: Melanoma is an aggressive skin cancer. Understanding the underlying mechanisms for melanomagenesis and identification of novel and effective melanoma treatment strategies are urgently necessary. The long-noncoding RNAs are considered as new essential players during cancer development, including the melanoma. Materials and methods: In this study, we first determined the expression of LINC00888 in tumor tissues and adjacent normal tissues from 28 patients with melanoma using quantitative polymerase chain reaction, and the correlation between the expression level of LINC00888 and the survival months was also examined...
2018: OncoTargets and Therapy
Tielei Tang, Yonggang Cheng, Qing She, Yaru Jiang, Yuanyuan Chen, Wenqiang Yang, Youhuai Li
Breast cancer is the leading cause of women death worldwide. Several long non-coding RNAs (lncRNAs) have been identified as oncogenes or tumor suppressors during the progression of cancers. However, the role of taurine upregulated gene (TUG1) in mediating the chemotherapy sensitivity of triple negative breast cancer (TNBC) has not been studied yet. In TNBC patients, we observed a significant decrease of TUG1 in tumor tissues compared to the normal tissues. Similarly, TUG1 expression was significantly decreased in TNBC cell lines compared with normal breast epithelial cell line and cell lines of other subtypes of breast cancer...
August 8, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Nannan Han, Hao Lu, Zun Zhang, Min Ruan, Wenjun Yang, Chenping Zhang
OBJECTIVES: To conduct an integrated analysis of microRNA and mRNA expression profile and further discover vital molecules to uncover novel pathogenic mechanisms in salivary adenoid cystic carcinoma (SACC). MATERIALS AND METHODS: MicroRNA and mRNA expression profiles were obtained from six paired primary SACC tumors and corresponding adjacent normal glands using high-throughput next-generation sequencing technology followed by an overall integrated bioinformatics analysis and subsequently molecular biology techniques validation...
August 8, 2018: Gene
Ming Bai, Heping Yin, Jian Zhao, Yang Li, Yimin Wu
BACKGROUND: Human bone marrow mesenchymal stem cells (hBM-MSC) have the ability of differentiating into chondrocytes and osteoblasts. miR-182-5p promotes osteoclastogenesis and bone metastasis by up-regulating the expression of parathyroid hormone-like hormone (PTHLH). However, the function of miR-182-5p in chondrogenesis is still unknown. METHODS: Mimic or inhibitor of miR-182-5p were used to upregulate or knock-down miR-182-5p expression respectively. We analyzed chondrogenesis by Safranin O staining and Blyscan™ Sulfated Glycosaminoglycan Assay...
August 10, 2018: Cell Biology International
Yuxin Xian, Liping Dong, Yong Jia, Yi Lin, Wenjuan Jiao, Yao Wang
miRNAs expression profiles in podocyte injuries have been reported in different models in vivo and in vitro. In the present study, miR-370 was elevated in high glucose-stimulated podocyte, and the post-transcriptional mechanism of miR-370 was investigated in high glucose-induced podocyte injuries. The gene and protein levels were assayed by RT-qPCR and western blotting, respectively. Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining was used to evaluate the apoptosis in high glucose-stimulated podocyte...
August 10, 2018: Cell Biology International
Yu-Liang Jiang, Zi-Ye Zhao, Bai-Rong Li, Fu Yang, Jing Li, Xiao-Wei Jin, Hao Wang, En-Da Yu, Shu-Han Sun, Shou-Bin Ning
BACKGROUND: Peutz-Jeghers syndrome (PJS) is caused by mutations in serine/threonine kinase 11 (STK11) gene. The increased cancer risk has been connected to P53 pathway. METHODS: PJS probands with STK11 mutation were included in the function analysis. P53 activity elevated by STK11 mutants was investigated using dual-luciferase reporter assay in vitro after constructing expression vectors of STK11 wild type and mutants generated by site-directed substitution. The association between the P53 activity and clinicopathological factors was analysis, especially the cancer history...
August 9, 2018: BMC Medical Genetics
M D Chanyshev, Y V Razumova, V Y Ovchinnikov, L F Gulyaeva
AIMS: The purpose of the present study was to determine whether miR-21 regulates the human ACAT1 gene. We also assessed whether transfection of MCF-7 cells with miR-21 mimic/inhibitor leads to changes in ACAT1 mRNA/protein levels, cell proliferation rate, or apoptosis. MAIN METHODS: Regulation of ACAT1 3'UTR by miR-21 was evaluated using a dual-luciferase reporter assay. The effect of miR-21 on mRNA/protein levels of ACAT1 and PTEN (confirmed as an important target of miR-21 for comparison) was measured by qPCR/western blot analysis and immunostaining...
August 6, 2018: Life Sciences
Xiu Qing Lin, Zhi Ming Huang, Xin Chen, Fang Wu, Wei Wu
PURPOSE: The influence of X-inactive specific transcript (XIST) and X-chromosome inactivation associated long non-coding RNAs (lncRNAs) just proximal to XIST (JPX) on hepatocellular carcinoma (HCC) remains controversial in light of previous reports, which the present study aimed to verify. MATERIALS AND METHODS: The DIANA lncRNA-microRNA (miRNA) interaction database was used to explore miRNA interactions with JPX or XIST. JPX, XIST, and miR-155-5p expression levels in paired HCC specimens and adjacent normal tissue were analyzed by RT-qPCR...
September 2018: Yonsei Medical Journal
Tao Zhang, Shuangfei Ni, Zixiang Luo, Ye Lang, Jianzhong Hu, Hongbin Lu
STUDY DESIGN: Experimental animal study. OBJECTIVES: To validate the anti-apoptosis effect of microRNA-21 in neurons after spinal cord injury (SCI) and explore the mechanism. SETTING: Xiangya Hospital, Central South University, Changsha, Hunan, People's Republic of China. METHODS: In situ hybridization was used to detect the expression of miR-21 in spinal cord neurons (n = 24). In a rat contusion SCI model (n = 48), we upregulated the miR-21 level around the injured area using miR-21 lentiviral vectors and evaluated the therapeutic effect with histology and behavioural scores...
August 8, 2018: Spinal Cord
Lina Wu, Zhen Chen, Yiqiao Xing
MicroRNAs (miRs) have been reported to participate in the initiation and progression of retinoblastoma (RB), as the most common primary intraocular malignancy during infancy and childhood worldwide. Recently, miR-506-3p has been reported to be associated with the development of RB, but the expression patterns and biological roles of miR-506-3p in RB have not been studied. In the present study, we found that miR-506-3p was significantly down-regulated in RB tissues and cell lines. Dual-luciferase reporter assay showed that miR-506-3p directly targeted mitosis Gene A (NIMA)-related kinase 6 (NEK6) in RB cells...
August 6, 2018: Cell Biology International
Fan Mao, Yue Lin, Yingli Zhou, Zhiying He, Jun Li, Yang Zhang, Ziniu Yu
Interferon regulatory factors (IRF), a family of transcription factors, are involved in the regulation of interferon to response the pathogen infection. Here, three IRF-like genes including CgIRF1a, CgIRF1b and CgIRF8 were identified in the genome of the oyster C. gigas. Among these genes, CgIRF1a and CgIRF1b, which are tandemly located in adjacent loci of scaffold 4, share the same domains. Phylogenetic analysis indicated that CgIRF1a and CgIRF1b were two paralogs that may originate from duplication of the same ancestral IRF gene...
August 2, 2018: Developmental and Comparative Immunology
Sheng-Hua Li, Jin-Pin Li, Lan Chen, Jing-Li Liu
Aberrant expression of miR-146a has been reported to be involved in the progression and metastasis of various types of human cancers; however, its potential role in human neuroblastoma is still poorly understood. The purpose of our study was to investigate the molecular mechanism and possible role of miR-146a in human neuroblastoma. In this study, targeted genes were predicted by bioinformatic analysis and confirmed by dual-Luciferase reporter assay. The expression level of miR-146a in the human neuroblastoma SK-N-SH cell line was detected by quantitative RT-PCR...
August 2018: Medical Hypotheses
Dong Ren, Hao Zheng, Sang Fei, Jia-Li Zhao
Long noncoding RNAs (LncRNAs) have been reported to participate in cancer development, including osteosarcoma. Here, in our study, we observed that lncRNA metastasis-associated lung adenocarcinoma transcription 1 (MALAT1) was remarkably overexpressed in osteosarcoma. However, the role it plays in osteosarcoma proliferation mediated by miR-206/cyclin-dependent kinase 9 (CDK9) axis remains uninvestigated. It was found that miR-206 was decreased and CDK9 was elevated in human osteosarcoma cells including MG63, Saos-2, U2OS, and KHOS compared with human osteoblast cell line hFOB 1...
August 4, 2018: Journal of Cellular Physiology
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