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dual luciferase reporter gene assay

Rui Zhou, Long Yi, Xikun Ye, Xianglong Zeng, Kai Liu, Yu Qin, Qianyong Zhang, Mantian Mi
BACKGROUND/AIMS: Lipid droplets (LDs) are dynamic organelles that store neutral lipids during times of energy excess, and an increased accumulation of LDs in the liver is closely linked to hepatic steatosis. Our previous studies suggested that resveratrol (RSV) supplement could improve hepatic steatosis, but the underlying mechanism, particularly which related to LD accumulation, has not yet been elucidated. METHODS: A high-fat diet (HFD) and palmitic acid were used to induce hepatic steatosis in mouse liver and hepatocytes, respectively...
2018: Cellular Physiology and Biochemistry
Lei Zhou, Li Yang, Yu-Jin Li, Rong Mei, Hua-Lin Yu, Yi Gong, Ming-Yue Du, Fei Wang
BACKGROUND/AIMS: Parkinson's disease (PD) is a frequently occurring condition that resulted from the loss of midbrain neurons, which synthesize the neurotransmitter dopamine. In this study, we established mouse models of PD to investigate the expression of microRNA-128 (miR-128) and mechanism through which it affects apoptosis of dopamine (DA) neurons and the expression of excitatory amino acid transporter 4 (EAAT4) via binding to axis inhibition protein 1 (AXIN1). METHODS: Gene expression microarray analysis was performed to screen differentially expressed miRNAs that are associated with PD...
2018: Cellular Physiology and Biochemistry
Yanwei Yin, Lei Du, Xuezhen Li, Xiaoyan Zhang, Yongli Gao
OBJECTIVE: This study aimed to investigate the expression levels of miR-133a-3p and collagen type I α 1 (COL1A1) in esophageal squamous cell carcinoma (ESCC) to find out the relationship between miR-133a-3p and COL1A1 and their influence on ESCC propagation, migration, invasion, and apoptosis. METHODS: The messenger RNA expression levels of miR-133a-3p and COL1A1 in ESCC were detected by quantitative reverse-transcription polymerase chain reaction. The expression of COL1A1 protein was examined via western blot analysis and immunohistochemistry assay...
December 10, 2018: Journal of Cellular Physiology
H-H Sun, P-F Sun, W-Y Liu
OBJECTIVE: This study aims to investigate whether miR-98-5p can participate in the myocardial differentiation of bone marrow mesenchymal stem cells (MSCs) by regulating TBX5. MATERIALS AND METHODS: In this study, we first identified the MSCs that were isolated from rat bone marrow samples. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to detect mRNA expressions of cardiac-related genes, including brain natriuretic peptide (BNP), α-actinin, and Islet-1...
November 2018: European Review for Medical and Pharmacological Sciences
X Luo, Y Song, L Tang, D-H Sun, D-G Ji
OBJECTIVE: To elucidate the biological functions of long non-coding RNA (lncRNA) SNHG7 in breast cancer (BC), and its underlying mechanism in the occurrence and progression of BC. PATIENTS AND METHODS: The expression of SNHG7 in 72 pairs of BC tissues and paracancerous tissues was detected by quantitative Real-time polymerase chain reaction (qRT-PCR). Correlation between SNHG7 expressions with pathological indicators of BC patients was analyzed. Similarly, SNHG7 expression in BC cell lines was determined by qRT-PCR as well...
November 2018: European Review for Medical and Pharmacological Sciences
H-E Zheng, G Wang, J Song, Y Liu, Y-M Li, W-P Du
OBJECTIVE: Different effects of microRNA-495 (miR-495) on human cancers have been exhibited in recent years. However, the specific function of miR-495 remains uncertain in non-small-cell lung cancer (NSCLC). Thus, we aim to explore the role of miR-495 in NSCLC. PATIENTS AND METHODS: The expressions of miR-495 and transcription factor 4 (TCF4) were detected through quantitative Real-time polymerase chain reaction (qRT-PCR) assay. Western blot was used to measure the protein expression of relative genes...
November 2018: European Review for Medical and Pharmacological Sciences
C-L Liu, W-H Wang, Y-L Sun, H-W Zhuang, M Xu, H-F Chen, J-X Liu
OBJECTIVE: The aim of this study was to investigate the role of miR-144-3p in the proliferation and metastasis capacity of pediatric Wilms' tumor (WT) cells and to explore the underlying mechanism. PATIENTS AND METHODS: The quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was performed to measure the expression level of miR-144-3p in pediatric WT tissues and cell lines (G401). A bioinformatics software was utilized to predict the interaction between miR-144-3p and Girdin...
November 2018: European Review for Medical and Pharmacological Sciences
Rong Jin, Sujuan Hu, Xiaomei Liu, Renzheng Guan, Ling Lu, Rongjun Lin
Asthma is a common chronic inflammatory respiratory disease characterised by airway inflammation and hyperresponsiveness. The present study was designed to clarify the effect of intranasal miR‑410 administration in an ovalbumin (OVA)‑induced murine model of asthma. It was found that miR‑410 expression was significantly decreased in the lungs of OVA‑induced asthmatic mice (P<0.05) and miR‑410 was overexpressed via intranasal instillation. Bioinformatics indicated that the 3'‑untranslated regions of interleukin (IL)‑4 and IL‑13 messenger RNAs (mRNAs) contain miR‑410 binding sites...
November 28, 2018: Molecular Medicine Reports
Masataka Shimonosono, Tetsuya Idichi, Naohiko Seki, Yasutaka Yamada, Takayuki Arai, Takaaki Arigami, Ken Sasaki, Itaru Omoto, Yasuto Uchikado, Yoshiaki Kita, Hiroshi Kurahara, Kosei Maemura, Shoji Natsugoe
Although miR‑145‑5p (the guide strand of the miR‑145 duplex) is established as a tumor suppressive microRNA (miRNA or miR), the functional significance of miR‑145‑3p (the passenger strand of the miR‑145 duplex) in cancer cells and its targets remains obscure. In our continuing analysis of esophageal squamous cell carcinoma (ESCC) pathogenesis, the aim of the present study was to identify important oncogenes and proteins that are controlled by miR‑145‑3p. Overexpression of miR‑145‑3p significantly reduced cancer cell proliferation, migration and invasive abilities, and further increased apoptotic abilities...
December 6, 2018: International Journal of Oncology
Nanpeng Li, Daojiang Li, Yuheng Du, Chen Su, Chunxing Yang, Changwei Lin, Xiaorong Li, Gui Hu
Researchers hold the view that PLAGL2 is overexpressed in many malignancies and that it can promote tumor proliferation, migration, invasion and self‑renewal; however, there is no evidence revealing a relationship between PLAGL2 and colorectal cancer (CRC). In the present study, genes that are overexpressed in CRC were screened using the COSMIC database and GEPIA database and the expression of PLAGL2 in carcinoma tissues and pericarcinomatous tissues was detected by RT‑qPCR and western blot assays. A Cell Counting Kit‑8 assay, a cell cycle analysis experiment and a xenograft model were used to explore the influence of PLAGL2 on CRC after knocking down PLAGL2 expression in HCT116 and SW480 cells...
December 7, 2018: Oncology Reports
Alok Khanduri, Amit Ranjan Sahu, Sajad Ahmad Wani, Raja Ishaq Nabi Khan, Aruna Pandey, Shikha Saxena, Waseem Akram Malla, Piyali Mondal, Kaushal Kishor Rajak, D Muthuchelvan, Bina Mishra, Aditya P Sahoo, Yash Pal Singh, Raj Kumar Singh, Ravi Kumar Gandham, Bishnu Prasad Mishra
In this study, the miRNAome and proteome of virulent Peste des petits ruminants virus (PPRV) infected goat peripheral blood mononuclear cells (PBMCs) were analyzed. The identified differentially expressed miRNAs (DEmiRNAs) were found to govern genes that modulate immune response based on the proteome data. The top 10 significantly enriched immune response processes were found to be governed by 98 genes. The top 10 DEmiRNAs governing these 98 genes were identified based on the number of genes governed by them...
2018: Frontiers in Immunology
Yunsheng Xu, Qin He, Yiyi Lu, Fengxing Tao, Liang Zhao, Rongying Ou
Background: We are committed to investigate miR-218-5 effects on the progression of cervical cancer (CC) cell and find out the molecular mechanism. Methods: GSE9750 was obtained from GEO database and R Limma package was applied to filter out dysregulated genes. The pathways were enriched by GSEA software, ClusterProfiler and enrichplot packages to predict the function of DEGs. The binding sites of LYN were detected by miRanda and TargetScan. The miR2Disease database was used to find miRNAs related with CC...
2018: Cancer Cell International
Jun-Feng Zhang, Jian-Shui Zhang, Zhao-Hua Zhao, Peng-Bo Yang, Sheng-Feng Ji, Nan Li, Qin-Dong Shi, Jing Tan, Xi Xu, Cang-Bao Xu, Ling-Yu Zhao
Background: MicroRNAs play crucial roles in tumorigenesis and tumor progression. miR-770 has been reported to be downregulated in several cancers and affects cancer cell proliferation, apoptosis, metastasis and drug resistance. However, the role and underlying molecular mechanism of miR-770 in human glioma remain unknown and need to be further elucidated. Methods: The expression of miR-770 in glioma tissues and cell lines was measured by quantitative real-time PCR (qRT-PCR) to explore the association of miR-770 expression with clinicopathological characteristics...
2018: Cancer Cell International
Qing Gao, Lin Xu, Qian Yang, Tian-Jun Guan
Peritoneal fibrosis remains to be one of the most severe causes of failure in continuous peritoneal dialysis. The current study cultured peritoneal mesothelial cells in high glucose to stimulate the environment of peritoneal fibrosis model in rats, and investigate whether microRNA-21 (miR-21) targeting Sprouty-1 affects high glucose-induced fibrosis in peritoneal mesothelial cells via the rennin angiotensin system (Ras)-mitogen-activated protein kinase (MAPK) signaling pathway, as well as potential mechanisms...
December 4, 2018: Journal of Cellular Physiology
Ying Yang, Bo Gong, Zheng-Zheng Wu, Ping Shuai, Dong-Feng Li, Ling-Lin Liu, Man Yu
Existing evidence has highlighted the effect of ultraviolet light radiation leading to corneal epithelium impairment. During this study, we aim to investigate the effect of microRNA-129-5p (miR-129-5p) on the wound healing process of corneal epithelial cells (CECs) induced by ultraviolet rays in mice by targeting epidermal growth factor receptor (EGFR). First, mouse models of ultraviolet ray-induced CEC injury were established and intrastromally injected with different mimic, inhibitor, and short interfering RNA (siRNA) to detect the effect of miR-129-5p on CEC injury...
December 4, 2018: Journal of Cellular Physiology
Dong-Mei Wu, Shan Wang, Xin Wen, Xin-Rui Han, Yong-Jian Wang, Shao-Hua Fan, Zi-Feng Zhang, Qun Shan, Jun Lu, Yuan-Lin Zheng
AIMS: We aimed to explore the impact of long noncoding RNA (lncRNA) nuclear enriched abundant transcript 1 (NEAT1) on cell proliferation, invasion, and migration of glioma. METHODS: Differentially expressed genes were screened out from Gene Expression Omnibus data set based on the microarray analysis. The expression levels of lncRNA NEAT1, miR-139-5p, and CDK6 in glioma cells and tissues were examined by quantitative reverse transcription polymerase chain reaction, and the protein level of CDK6 in glioma cells was determined by western blot and immunohistochemistry...
December 4, 2018: Journal of Cellular Physiology
Xi Sun, Yue Hu, Jing Wu, Liang Shi, Lei Zhu, Pei-Wen Xi, Ji-Fu Wei, Qiang Ding
BACKGROUND: RNA binding proteins (RBPs) play an important role in regulating the metabolism of target RNAs. Aberrant expression of RBPs plays a vital role in the initiation and development of many cancers. The RBM family, which has the conserved RNA binding motif RNP1 and RNP2, shares the similar function in RNA processing and RBMS2 is a member of them. P21, also named CDKN1A, promotes cell cycle arrest and plays an important role in halting cell proliferation. In our study, we identified RBMS2 as a tumor suppressor in breast cancer...
December 4, 2018: Journal of Experimental & Clinical Cancer Research: CR
Lei Yang, Dawei Ge, Xi Chen, Chunzhi Jiang, Shengnai Zheng
BACKGROUND/AIMS: To explore the potential role of miR-544a in spinal cord injury and the possible mechanism involved. METHODS: We established a mouse model with spinal cord injury to examine the changes in grip force recovery of the forelimb or the posterior limb of the mouse. Microarray was performed to achieve differentiated miRNAs in the mice. The expressions of miR-544a, MCP-1, IL36B and IL17B after spinal cord injury were detected by qRT-PCR. Subsequently, miR-544a was overexpressed to observe changes in inflammation and grip strength after spinal cord injury...
December 4, 2018: Cellular Physiology and Biochemistry
Handong Sun, Pengcheng Xi, Zhiqiang Sun, Qian Wang, Bin Zhu, Jian Zhou, Hui Jin, Wubin Zheng, Weiwei Tang, Hongyong Cao, Xiufeng Cao
Background: Circular RNAs(circRNAs) have been reported as a diverse class of endogenous RNA that regulate gene expression in eukaryotes. Recent evidence suggested that many circular RNAs can act as oncogenes or tumor suppressors through sponging microRNAs. However, the function of circular RNAs in gastric cancer remains largely unknown. Materials and methods: The circRNA levels in gastric carcinoma tissues and plasmas were detected by real-time quantitative reverse transcription-polymerase chain reaction...
2018: Cancer Management and Research
Y Mi, F Liu, X Liang, S Liu, X Huang, M Sang, C Geng
Let-7 was one of the earliest discovered miRNAs and while it reportedly acts as a tumor suppressor in various solid tumors, its function in breast cancer has not been fully studied. Therefore, we examined let-7a and MAGE-A1 expression in breast tissues by qRT-PCR and found that let-7a expression significantly correlates with larger tumor size, higher histological grade (P < 0.05) and is significantly lower in patients with Her-2-positive cancers and Ki-67 > 14% (P = 0.028 and P = 0.023). MAGE-A1 expression incidence is 50...
August 9, 2018: Neoplasma
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