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dual luciferase reporter gene assay

Ke Xu, Bin Han, Yang Bai, Xiu-Ying Ma, Zhen-Ni Ji, Yao Xiong, Shi-Kun Miao, Yuan-Yuan Zhang, Li-Ming Zhou
The global morbidity and mortality of colorectal cancer (CRC) are ranked the third among gastrointestinal tumors in the world. MiR-451a is associated with several types of cancer, including CRC. However, the roles and mechanisms of miR-451a in CRC have not been elucidated. BAP31 is a predicted target gene of miR-451a in our suppression subtractive hybridization library. Its relationship with miR-451a and function in CRC are unclear. We hypothesized that miR-451a could induce apoptosis through suppressing BAP31 in CRC...
February 15, 2019: Cell Death & Disease
He Hu, Chuanlong Zhao, Peiguang Zhang, Yalong Liu, Yulian Jiang, Enquan Wu, Hao Xue, Caiyun Liu, Zhehai Li
BACKGROUNDS: Osteoactivin (OA) is a key regulator promoting bone marrow stromal cells osteogenesis progress, while Dexamethasone (Dex) could inhibit OA induced osteogenesis and lead to osteoporosis. miR-26b increased during BMSC osteogenesis but whether it participates in this progress is enigma. Osteogenesis is under regulation of canonical Wnt signaling pathway which could serve as potential target for miR-26b. It bears therapeutic potential if miR-26b could regulate osteogenesis and antagonize Dex induced Osteoporosis (OP)...
February 12, 2019: Experimental and Molecular Pathology
Xue Wu, Mei Ding, Yi Liu, Xi Xia, Feng-Ling Xu, Jun Yao, Bao-Jie Wang
Abnormal expression of the 5-HT1A receptor, which is encoded by the HTR1A gene, leads to susceptibilities to neuropsychiatric disorders such as depression, anxiety, and schizophrenia. miRNAs regulate gene expression by recognizing the 3'-UTR region of mRNA. This study evaluated the miRNAs that might identify and subsequently determine the regulatory mechanism of HTR1A gene. Using the HEK-293, U87, SK-N-SH and SH-SY5Y cell lines, we determined the functional sequence of the 3'-UTR region of the HTR1A gene and predicted miRNA binding...
2019: Frontiers in Molecular Neuroscience
Zengkai Zhang, Yujun Chen, Bojiang Li, Yilong Yao, Aiwen Jiang, Wei Wei, Honglin Liu, Wangjun Wu
MicroRNAs (miRNAs) are endogenous short non-coding RNAs and exert their function by targeting mRNAs of genes. miRNA-206 (miR-206) is exclusively expressed in adult skeletal muscles and plays an important role in myogenesis. However, the regulatory mechanisms of miR-206 in myoblasts proliferation and differentiation are still limited. In this study, we validated that Notch3 is a direct target gene of mouse miR-206 using dual-luciferase reporter assay and miR-206-overexpressed experiments. Furthermore, we demonstrated that the negative effect of overexpression of miR-206 and knockdown of Notch3 on C2C12 cells proliferation, and found that the effect was produced by controlling the transition of G0/G1 and S phase in C2C12 cell cycle...
February 11, 2019: Gene
Ruinan Shen, Suzhen Lin, Lu He, Xue Zhu, Zhekun Zhou, Shengdi Chen, Ying Wang, Jianqing Ding
Background: Parkinson's disease (PD) is the most common neurodegenerative movement disorder that is known to be related to neuro-inflammation. Chemokines participate in this process usually through upregulation of expression levels, which are closely related to the polymorphisms in their genes. Recent studies have further revealed the association between these polymorphisms and the risk of PD in multiple populations, but not the Chinese Han population. Methods: The promoter region of CCL2 was sequenced in 411 PD patients and 422 gender-age matched control from a Chinese Han population using PCR-RFLP method...
2019: Frontiers in Neurology
Peng Luo, Qi Wang, Yuanyuan Ye, Ju Zhang, Dapeng Lu, Longqiang Cheng, Hangcheng Zhou, Mingran Xie, Baolong Wang
BACKGROUND: MicroRNAs have an important role in diverse biological processes including tumorigenesis. MiR-223 has been reported to be deregulated in several human cancer types. However, its biological role has not been functionally characterized in lung squamous cell carcinoma (LSCC). The following study investigates the role of miR-223-3p in LSCC growth and metastasis and its underlying mechanism. METHODS: MicroRNA profiling analyses were conducted to determine differential miRNAs expression levels in LSCC tumor tissues that successfully formed xenografts in immunocompromised mice (XG) and failed tumor tissues (no-XG)...
February 12, 2019: Journal of Experimental & Clinical Cancer Research: CR
Bo-Yu Yang, Guo-Ying Deng, Rui-Zhe Zhao, Chen-Yun Dai, Chen-Yi Jiang, Xing-Jie Wang, Yi-Feng Jing, Xi-Jian Liu, Shu-Jie Xia, Bang-Min Han
Benign prostatic hyperplasia (BPH) patients experience complications after surgery. We studied oxidative stress scavenging by porous Se@SiO2 nanospheres in prostatic urethra wound healing after transurethral resection of the prostate (TURP). Beagle dogs were randomly distributed into two groups after establishing TURP models. Wound recovery and oxidative stress levels were evaluated. Re-epithelialization and the macrophage distribution at the wound site were assessed by histology. The mechanism by which porous Se@SiO2 nanospheres regulated macrophage polarization was investigated by qRT-PCR, western blotting, flow cytometry, immunofluorescence and dual luciferase reporter gene assays...
February 9, 2019: Acta Biomaterialia
Yueyun Ding, Shujiao Zhu, Chaodong Wu, Li Qian, Dengtao Li, Li Wang, Yuanlang Wang, Wei Zhang, Min Yang, Jian Ding, Xudong Wu, Xiaodong Zhang, Yafei Gao, Zongjun Yin
Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a...
January 3, 2019: Asian-Australasian Journal of Animal Sciences
Yanmei Jia, Lirong Chen, Songjia Guo, Yuanhong Li
To analyze the anti-tumor mechanism of Baicalin in human colon cancer. The MTT assay and colony formation assay demonstrated that Baicalin treatment inhibits the proliferation of DLD1 and HCT-116 cells. The apoptosis rates were induced upon Baicalin treatment and which was determined by FACS. The qPCR and western blot analysis showed that Baicalin promotes expression of DKK1 (Dickkopf), an important antagonist of Wnt signaling pathway, thereby reduces the expression of its downstream protein β-catenin and c-Myc...
February 8, 2019: Molecular Biology Reports
Zhenwei He, Xuelei Ruan, Xiaobai Liu, Jian Zheng, Yunhui Liu, Libo Liu, Jun Ma, Lianqi Shao, Di Wang, Shuyuan Shen, Chunqing Yang, Yixue Xue
BACKGROUND: Angiogenesis plays a critical role in the progression of glioma. Previous studies have indicated that RNA-binding proteins (RBPs) interact with RNAs and participate in the regulation of the malignant behaviors of tumors. As a type of endogenous non-coding RNAs, circular RNAs (circRNAs) are abnormally expressed in various cancers and are involved in diverse tumorigeneses including angiogenesis. METHODS: The expression levels of FUS, circ_002136, miR-138-5p, SOX13, and SPON2 were determined using quantitative real-time PCR (qRT-PCR) and western blot...
February 8, 2019: Journal of Experimental & Clinical Cancer Research: CR
Ting Yuan, Zhen Ni, Chuan Han, Yali Min, Nina Sun, Caifang Liu, Miao Shi, Wenquan Lu, Na Wang, Feng Du, Qiong Wu, Ning Xie, Yongquan Shi
Background: Intestinal metaplasia (IM) is a premalignant lesion associated with gastric cancer. Both animal and clinical studies have revealed that bile acid reflux and subsequent chronic inflammation are key causal factors of IM. Previous studies indicated that SOX2, the key transcription factor in gastric differentiation, was downregulated during IM development while CDX2, the pivotal intestine-specific transcription factor was upregulated significantly. However, it remains unclear whether the downregulation of SOX2 promotes gastric IM emergence or is merely a concomitant phenomenon...
2019: Cancer Cell International
Q Wu, D-Z Xi, Y-H Wang
OBJECTIVE: This study investigates whether microRNA-599 can inhibit the progression of Parkinson's disease (PD) by regulating the LRRK2 expression. We aim to search for a new therapeutic target for PD. MATERIALS AND METHODS: A mouse model of PD was first established. A relative amount of TH+ neurons in the mouse brain was quantified by immunohistochemistry. The expression levels of microRNA-599 and LRRK2 in mouse brain tissues were determined by the quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot...
January 2019: European Review for Medical and Pharmacological Sciences
C-M Xu, L-X Chen, F Gao, M-F Zhu, Y Dai, Y Xu, W-X Qian
OBJECTIVE: We aimed to detect the role and function of microRNA-431 (miR-431) in lung cancer, and to investigate the underlying mechanism in regulating the development of lung cancer. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was utilized to measure the relative expression level of miR-431 in lung cancer tissues and cell lines. Cell counting kit-8 (CCK-8) and colony formation assays were employed to measure the proliferative ability of lung cancer cells...
January 2019: European Review for Medical and Pharmacological Sciences
W-G Bian, X-N Zhou, S Song, H-T Chen, Y Shen, P Chen
OBJECTIVE: Accumulating evidence has suggested that aberrant expression of microRNAs (miRNAs) is associated with non-small cell lung cancer (NSCLC) proliferation, migration, invasion and chemotherapy resistance. Cullin4A (CUL4A) has been previously reported to desensitize NSCLC cells to chemotherapy treatment. However, whether miRNAs regulate CUL4A to promote chemotherapy resistance remains unknown. PATIENTS AND METHODS: Tissues were obtained from 40 NSCLC patients who received surgery at the Yancheng City No...
January 2019: European Review for Medical and Pharmacological Sciences
Y-M Du, Y-B Wang
OBJECTIVE: This study aimed at investigating whether miR-637 could promote proliferation of hepatocarcinoma cells by targeted regulation of AKT1 expression, leading to enhanced cell invasion and thus participating in the progression of liver cancer. PATIENTS AND METHODS: The miR-637 and AKT1 expressions in cancer tissues and adjacent tissues of liver cancer patients were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The effects of miR-637 on cell proliferation and cell invasion were examined by cell counting kit-8 (CCK-8) and cell invasion assays...
January 2019: European Review for Medical and Pharmacological Sciences
M Li, Y-L Zhang, H Huang, Y Xiong
OBJECTIVE: The purpose of the study was to investigate the regulatory effect of microRNA-10-5p on TBX5 during myocardial differentiation of bone marrow mesenchymal stem cells (BMSCs) in rats. MATERIALS AND METHODS: Rat BMSCs were first isolated, cultured and identified by flow cytometry. Expression levels of BNP (Brain Natriuretic Peptide), α-actinin, and Islet-1 in BMSCs co-cultured with rat cardiomyocytes were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR)...
January 2019: European Review for Medical and Pharmacological Sciences
H-G Zhang, X-B Wang, H Zhao, C-N Zhou
OBJECTIVE: To explore the role of microRNA-9-5p in regulating osteoporosis (OS) development and its underlying mechanism. PATIENTS AND METHODS: MicroRNA-9-5p expression in peripheral blood of 30 OS patients and 30 healthy subjects was examined by quantitative Real-Time-Polymerase Chain Reaction (qRT-PCR). During the processes of osteogenesis and adipogenesis, mRNA levels of microRNA-9-5p, osteogenesis-related genes, and adipogenesis-related genes in marrow stromal stem cells (MSCs) were detected by qRT-PCR as well...
January 2019: European Review for Medical and Pharmacological Sciences
Shan Xu, Lian Hui, Ning Yang, Yan Wang, Ning Zhao, Xue-Jun Jiang
Hypopharyngeal carcinoma (HPC) is an aggressive malignancy with the worst prognosis among all head and neck cancers. MicroRNAs (miRNAs) are involved in the development of many human cancers, and may function as oncogenes or tumor suppressors. The present study aimed to evaluate the effects of miRNA (miR)‑194‑5p on the proliferation and invasion of HPC cells and to identify the potential regulatory mechanism. First, miR‑194‑5p and Smad ubiquitin regulatory factor 1 (SMURF1) expression levels were examined in HPC tissues...
February 4, 2019: International Journal of Oncology
Xiao-Xue Wang, Heyang Zhang, Yan Li
MicroRNA (miR)‑148a is differentially expressed in numerous malignant tumors and it was identified to regulate tumor growth, cell proliferation, apoptosis, angiogenesis and drug resistance via the regulation of the expression levels of its target genes. However, the biological function of miR‑148a in acute myeloid leukemia (AML) and its molecular mechanisms of action remain unclear. In the present study, the expression levels of miR‑148a and DNA methyltransferase 1 (DNMT1) were detected using reverse transcription‑quantitative polymerase chain reaction (PCR) and western blotting...
January 30, 2019: Molecular Medicine Reports
Juan Wang, Pradhan Abhinav, Ying-Jia Xu, Ruo-Gu Li, Min Zhang, Xing-Biao Qiu, Ruo-Min Di, Qi Qiao, Xiu-Mei Li, Ri-Tai Huang, Song Xue, Yi-Qing Yang
Congenital bicuspid aortic valve (BAV) represents the most common type of cardiac birth defect affecting 0.4‑2% of the general population, and accounts for a markedly increased incidence of life‑threatening complications, including valvulopathy and aortopathy. Accumulating evidence has demonstrated the genetic basis of BAV. However, the genetic basis for BAV in the majority of cases remains to be elucidated. In the present study, the coding regions and splicing donors/acceptors of the nuclear receptor subfamily 2 group F member 2 (NR2F2) gene, which encodes a transcription factor essential for proper cardiovascular development, were sequenced in 176 unrelated cases of congenital BAV...
January 31, 2019: International Journal of Molecular Medicine
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