scholarly journals miR-19a-3p Promotes Tumor-Relevant Behaviors in Bladder Urothelial Carcinoma via Targeting THBS1

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Gang Xu ◽  
Junlong Li ◽  
Lihang Yu
Keyword(s):  
Epithelial Cells ◽  
Western Blot ◽  
Urothelial Carcinoma ◽  
Bioinformatics Analysis ◽  
Luciferase Gene ◽  
Real Time Quantitative Pcr ◽  
Functional Assays ◽  
Bladder Urothelial Carcinoma ◽  
Gene Assay ◽  
Cell Progression

Objective. miR-19a-3p is widely increased in several cancers and can be used as an oncogenic factor in these cancers. However, the molecular mechanism of miR-19a-3p in bladder urothelial carcinoma (BLCA) is still open. So, the study was aimed at exploring the mechanism of miR-19a-3p in BLCA cells. Methods. Bioinformatics analysis was employed to find the differential miRNAs and mRNAs, and the target miRNA and mRNA were determined. Real-time quantitative PCR was used to evaluate miR-19a-3p and THBS1 levels in human urethral epithelial cells and BLCA cells. Western blot was carried out to assay protein expression of THBS1 in human urethral epithelial cells and BLCA cells. Behaviors of BLCA cells were detected through cellular functional assays. Dual-luciferase gene assay was conducted to validate the binding of miR-19a-3p and THBS1. Results. miR-19a-3p was increased in BLCA cells, while THBS1 was less expressed in BLCA cells. The miR-19a-3p functions as an oncogene in BLCA. THBS1 was a target of miR-19a-3p, and it could reverse the promotion of miR-19a-3p on cell malignant behaviors in BLCA. Conclusion. miR-19a-3p facilitates cell progression in BLCA via binding THBS1, which may be an underlying therapeutic target for BLCA treatment.

scholarly journals Let-7c-3p Regulates Autophagy under Oxidative Stress by Targeting ATG3 in Lens Epithelial Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-9 ◽  
Author(s):  
Ting Li ◽  
Yanhong Huang ◽  
Wenkai Zhou ◽  
Qichang Yan
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Western Blot ◽  
Molecular Mechanisms ◽  
Luciferase Reporter ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Age Related ◽  
Spot Number ◽  
Gene Assay

Background. Oxidative stress is an important factor during age-related cataract formation. Apoptosis and autophagy induced by oxidative stress have been reported as key factors in age-related cataract. In our research, we investigated the role of let-7c-3p in the regulation of autophagy and apoptosis during the formation of age-related cataract. Material and Methods. Real-time PCR and western blot were employed to detect the expression of let-7c-3p in the tissues of age-related cataract. Human lens epithelial cells (LECs) were treated with H2O2 as an age-related cataract model. The extent of apoptosis was measured by flow cytometry and western blot. To detect autophagy, immunofluorescence was used to analyze the spot number of LC3, and western blot was used to detect the expression of LC3-II/I and ATG3. The molecular mechanisms of let-7c-3p regulating autophagy via ATG3 under oxidative stress were performed by a luciferase report gene assay and rescue experiment. Results. Downregulation of let-7c-3p was found in the age-related cataract group aged >65 years relative to the age-related cataract group aged ≤65 years. Consistently, the expression of let-7c-3p was also lower under oxidative stress. The activities of LEC apoptosis and autophagy induced by oxidative stress were inhibited by let-7c-3p. By the bioinformatics database and the luciferase reporter assay, ATG3 was found to be a direct target of let-7c-3p. Let-7c-3p reduced the ATG3-mediated autophagy level, which was induced by oxidative stress in LECs. Conclusion. Let-7c-3p inhibits autophagy by targeting ATG3 in LECs in age-related cataract.

scholarly journals Seasonal Expression of Extracellular Signal Regulated Kinases in the Colon of Wild Ground Squirrels (Spermophilus Dauricus)

2021 ◽  
Author(s):  
Yue Song ◽  
Xiaoying Yang ◽  
Xueying Zhang ◽  
Jueyu Zhu ◽  
Yixin Chen ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Western Blot ◽  
Breeding Season ◽  
Seasonal Changes ◽  
Intestinal Epithelial Cells ◽  
Goblet Cells ◽  
Ground Squirrels ◽  
Intestinal Epithelial ◽  
Real Time Quantitative Pcr ◽  
Extracellular Signal

Abstract Background: The aim of this study was to explore the localization and expression of extracellular signal regulated kinase/phospho-extracellular signal regulated kinases (ERK/pERK) in the colonic tissue of wild ground squirrels (Spermophilus dauricus) in the breeding season and the non-breeding season. Methods and Results: Hematoxylin-eosin staining, immunohistochemistry, real-time quantitative PCR and Western blot were used in this study. The histological results showed that the diameter of the colon lumen in the non-breeding season was larger than that in the breeding season, and the number of glandular cells in the non-breeding season was also more than those in the breeding season. The immunochemical results showed that ERK1/2 was expressed in the cytoplasm of goblet cells and intestinal epithelial cells, while pERK1/2 was expressed in the cytoplasm and nucleus of goblet cells and intestinal epithelial cells. The immunolocalization of ERK1/2 and pERK1/2 expression were more obvious in the non-breeding season, especially in intestinal epithelial cells. The results of real-time quantitative PCR and Western blot showed that the expression of ERK1/2 and pERK1/2 in the breeding season was significantly lower than that in the non-breeding season.Conclusions: The expression of ERK1/2 and pERK1/2 in the colons of the wild ground squirrels had seasonal changes, which had significant increases in the non-breeding season comparing to those in the breeding season. This study revealed the potential role of ERK1/2 in colon to the adaptation of seasonal changes in wild ground squirrel.

scholarly journals miR-9-5p Promotes Lung Adenocarcinoma Cell Proliferation, Migration and Invasion by Targeting ID4

2021 ◽  
Vol 20 ◽  
pp. 153303382110485
Author(s):  
Kai Zhu ◽  
Jinlan Lin ◽  
Shengjia Chen ◽  
Qian Xu
Keyword(s):  
Lung Adenocarcinoma ◽  
Regulatory Mechanism ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Reporter Assay ◽  
Biological Functions ◽  
Real Time Quantitative Pcr ◽  
Functional Assays ◽  
Cell Progression ◽  
Dual Luciferase Reporter Assay

Objectives Evidence reveals that microRNAs (miRNAs) are abnormally expressed in lung adenocarcinoma (LUAD) tissue and are crucial in LUAD occurrence. Therefore, this study aims to find the miRNA which could regulate LUAD and to further explore its regulatory mechanism, thus providing a potential molecular target for LUAD. Methods miR-9-5p and ID4 expression in LUAD cells were measured by real-time quantitative PCR and western blot. Cell functional assays were conducted to detect the biological functions of LUAD cells. A dual-luciferase reporter assay was utilized to validate the binding relationship between miR-9-5p and ID4. Results miR-9-5p was highly expressed whereas ID4 was lowly expressed in LUAD. miR-9-5p facilitated LUAD cell progression by targeting ID4. Conclusion miR-9-5p promotes LUAD cell progression by modulating ID4 and may become a potential target for LUAD.

scholarly journals Rspo3 regulates the abnormal differentiation of small intestinal epithelial cells in diabetic state

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ti-Dong Shan ◽  
Han Yue ◽  
Xue-Guo Sun ◽  
Yue-Ping Jiang ◽  
Li Chen
Keyword(s):  
Epithelial Cells ◽  
Bioinformatics Analysis ◽  
Intestinal Epithelial Cells ◽  
Underlying Mechanism ◽  
Luciferase Reporter ◽  
Intestinal Epithelial ◽  
Epithelial Stem Cells ◽  
Diabetic State ◽  
Definitive Evidence

Abstract Background The complications caused by diabetes mellitus (DM) are the focus of clinical treatment. However, little is known about diabetic enteropathy (DE) and its potential underlying mechanism. Methods Intestinal epithelial cells (IECs) and intestinal epithelial stem cells (IESCs) were harvested from BKS.Cg-Dock7m+/+Leprdb/JNju (DM) mice, and the expression of R-Spondin 3 (Rspo3) was detected by RT-qPCR, Western blotting, immunohistochemistry, and immunofluorescence. The role of Rspo3 in the abnormal differentiation of IECs during DM was confirmed by knockdown experiments. Through miRNA expression profiling, bioinformatics analysis, and RT-qPCR, we further analyzed the differentiation-related miRNAs in the IECs from mice with DM. Results Abnormal differentiation of IECs was observed in the mice with DM. The expression of Rspo3 was upregulated in the IECs from the mice with DM. This phenomenon was associated with Rspo3 overexpression. Additionally, Rspo3 is a major determinant of Lgr5+ stem cell identity in the diabetic state. Microarray analysis, bioinformatics analysis, and luciferase reporter assays revealed that microRNA (miR)-380-5p directly targeted Rspo3. Moreover, miR-380-5p upregulation was observed to attenuate the abnormal differentiation of IECs by regulating Rspo3 expression. Conclusions Together, our results provide definitive evidence of the essential role of Rspo3 in the differentiation of IECs in DM.

EMP2 induces cytostasis and apoptosis via the TGFβ/SMAD/SP1 axis and recruitment of P2RX7 in urinary bladder urothelial carcinoma

2021 ◽  
Author(s):  
Chien-Feng Li ◽  
Ti-Chun Chan ◽  
Cheng-Tang Pan ◽  
Pichpisith Pierre Vejvisithsakul ◽  
Jia-Chen Lai ◽  
...  
Keyword(s):  
Urinary Bladder ◽  
Urothelial Carcinoma ◽  
Bladder Urothelial Carcinoma
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