AIM:To study microRNAs(miRNAs)and their potential effects in high glucose-induced human retinal pigment epithelial cell damage.METHODS:We screened the GSE52233 miRNA expression dataset for differentially expressed miR...AIM:To study microRNAs(miRNAs)and their potential effects in high glucose-induced human retinal pigment epithelial cell damage.METHODS:We screened the GSE52233 miRNA expression dataset for differentially expressed miRNAs(DEMs).The target genes of the top 10 DEMs were predicted using miRWalk 2.0 database,followed by function enrichment and protein-protein interaction analysis.miRNA expression was determined in the human retinal pigment epithelial cell line ARPE-19 treated with high glucose(HG)by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Cell proliferation was determined using cell counting kit(CCK)-8 assay.Cell cycle,apoptosis,and reactive oxygen species(ROS)levels were determined by flow cytometry.The direct interaction between miRNA and targets was validated using dual-luciferase reporter assay.RESULTS:Thirty-nine DEMs were screened,and we predicted 125 miRNA-mRNA pairs for the top 10 DEMs,including 119 target genes of seven DEMs such as miR-346,which was upregulated in diabetic retinopathy(DR).miR-346 target genes were substantially enriched in the regulation of intracellular transport and retinoic acidinducible gene I(RIG-I)-like receptor signaling pathway.Expression of three upregulated and downregulated miRNAs were verified by qRT-PCR in HG-treated ARPE-19 cells.Expression of miR-346 was elevated in HG treated ARPE-19 cells in a dose-dependent manner.HG inhibited cell proliferation and induced apoptosis,which were partly reversed by transfecting an miR-346 inhibitor,which even decreased the ROS levels elevated due to HG.Argonaute 2(AGO2)was a target of miR-346.CONCLUSION:miR-346 is a key miRNA and plays an important role in HG-induced damage in human retinal pigment epithelial cells.展开更多
[目的]探究miR-346与TNF-α/Mfn2通路对脑梗死小鼠内质网应激和神经元凋亡的影响。[方法]将C57BL/6J小鼠随机分为3组:假手术组(sham)、NC agomir组、miR-346 agomir组。通过苏木素伊红染色分析海马组织病理变化,TUNEL实验检测神经细胞...[目的]探究miR-346与TNF-α/Mfn2通路对脑梗死小鼠内质网应激和神经元凋亡的影响。[方法]将C57BL/6J小鼠随机分为3组:假手术组(sham)、NC agomir组、miR-346 agomir组。通过苏木素伊红染色分析海马组织病理变化,TUNEL实验检测神经细胞凋亡率,通过氯化三苯基四氮唑染色评估小鼠的脑梗死面积,尼氏染色分析神经元尼氏小体数量,蛋白免疫印迹实验分析TNF-α/Mfn2通路以及内质网应激相关蛋白的表达水平。[结果]与假手术组的小鼠比较,NC agomir组的小鼠的脑梗死面积增加(0.21±0.01 vs 27.16±4.38,P<0.05),海马组织受到破坏,神经元尼氏小体的数量减少(56.27±5.13 vs 12.79±2.95,P<0.05),神经元凋亡率增加(1.25%±0.03%vs 17.89%±2.96%,P<0.05),内质网应激相关蛋白以及TNF-α表达增加(0.23±0.05 vs 1.22±0.13,P<0.05),Mfn2表达降低(0.97±0.06 vs 0.13±0.03,P<0.05)。与NC agomir组的小鼠比较,miR-346 agomir组的小鼠的脑梗死面积减少,海马组织结构得到改善,神经元尼氏小体的数量增加,神经元凋亡率降低,内质网应激相关蛋白以及TNF-α表达降低,Mfn2表达增加。[结论]miR-346能够调控TNF-α/Mfn2信号轴,减少脑梗死小鼠海马组织区域的内质网应激和神经元凋亡。展开更多
BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact mo...BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.展开更多
基金Supported by the Social Development Project of Shaanxi Provincial Department of Science and Technology(No.2020SF-167)Supporting Fund Project of Shaanxi Provincial Department of Science and Technology Agency Project(No.2022SF-502)Xi’an Medical University 2022 Annual Scientific Research Capacity Improvement Plan Project(No.2022NLTS104).
文摘AIM:To study microRNAs(miRNAs)and their potential effects in high glucose-induced human retinal pigment epithelial cell damage.METHODS:We screened the GSE52233 miRNA expression dataset for differentially expressed miRNAs(DEMs).The target genes of the top 10 DEMs were predicted using miRWalk 2.0 database,followed by function enrichment and protein-protein interaction analysis.miRNA expression was determined in the human retinal pigment epithelial cell line ARPE-19 treated with high glucose(HG)by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).Cell proliferation was determined using cell counting kit(CCK)-8 assay.Cell cycle,apoptosis,and reactive oxygen species(ROS)levels were determined by flow cytometry.The direct interaction between miRNA and targets was validated using dual-luciferase reporter assay.RESULTS:Thirty-nine DEMs were screened,and we predicted 125 miRNA-mRNA pairs for the top 10 DEMs,including 119 target genes of seven DEMs such as miR-346,which was upregulated in diabetic retinopathy(DR).miR-346 target genes were substantially enriched in the regulation of intracellular transport and retinoic acidinducible gene I(RIG-I)-like receptor signaling pathway.Expression of three upregulated and downregulated miRNAs were verified by qRT-PCR in HG-treated ARPE-19 cells.Expression of miR-346 was elevated in HG treated ARPE-19 cells in a dose-dependent manner.HG inhibited cell proliferation and induced apoptosis,which were partly reversed by transfecting an miR-346 inhibitor,which even decreased the ROS levels elevated due to HG.Argonaute 2(AGO2)was a target of miR-346.CONCLUSION:miR-346 is a key miRNA and plays an important role in HG-induced damage in human retinal pigment epithelial cells.
文摘[目的]探究miR-346与TNF-α/Mfn2通路对脑梗死小鼠内质网应激和神经元凋亡的影响。[方法]将C57BL/6J小鼠随机分为3组:假手术组(sham)、NC agomir组、miR-346 agomir组。通过苏木素伊红染色分析海马组织病理变化,TUNEL实验检测神经细胞凋亡率,通过氯化三苯基四氮唑染色评估小鼠的脑梗死面积,尼氏染色分析神经元尼氏小体数量,蛋白免疫印迹实验分析TNF-α/Mfn2通路以及内质网应激相关蛋白的表达水平。[结果]与假手术组的小鼠比较,NC agomir组的小鼠的脑梗死面积增加(0.21±0.01 vs 27.16±4.38,P<0.05),海马组织受到破坏,神经元尼氏小体的数量减少(56.27±5.13 vs 12.79±2.95,P<0.05),神经元凋亡率增加(1.25%±0.03%vs 17.89%±2.96%,P<0.05),内质网应激相关蛋白以及TNF-α表达增加(0.23±0.05 vs 1.22±0.13,P<0.05),Mfn2表达降低(0.97±0.06 vs 0.13±0.03,P<0.05)。与NC agomir组的小鼠比较,miR-346 agomir组的小鼠的脑梗死面积减少,海马组织结构得到改善,神经元尼氏小体的数量增加,神经元凋亡率降低,内质网应激相关蛋白以及TNF-α表达降低,Mfn2表达增加。[结论]miR-346能够调控TNF-α/Mfn2信号轴,减少脑梗死小鼠海马组织区域的内质网应激和神经元凋亡。
基金Supported by the Haihe Laboratory of Cell Ecosystem Innovation Fund,No.22HHXBJC00001the Key Discipline Special Project of Tianjin Municipal Health Commission,No.TJWJ2022XK016.
文摘BACKGROUND Hepatocellular carcinoma(HCC)has been a pervasive malignancy throughout the world with elevated mortality.Efficient therapeutic targets are beneficial to treat and predict the disease.Currently,the exact molecular mechanisms leading to the progression of HCC are still unclear.Research has shown that the microRNA-142-3p level decreases in HCC,whereas bioinformatics analysis of the cancer genome atlas database shows the ASH1L expression increased among liver tumor tissues.In this paper,we will explore the effects and mechanisms of microRNA-142-3p and ASH1L affect the prognosis of HCC patients and HCC cell bioactivity,and the association between them.AIM To investigate the effects and mechanisms of microRNA-142-3p and ASH1L on the HCC cell bioactivity and prognosis of HCC patients.METHODS In this study,we grouped HCC patients according to their immunohistochemistry results of ASH1L with pathological tissues,and retrospectively analyzed the prognosis of HCC patients.Furthermore,explored the roles and mechanisms of microRNA-142-3p and ASH1L by cellular and animal experiments,which involved the following experimental methods:Immunohistochemical staining,western blot,quantitative real-time-polymerase chain reaction,flow cytometric analysis,tumor xenografts in nude mice,etc.The statistical methods involved in this study contained t-test,one-way analysis of variance,theχ^(2)test,the Kaplan-Meier approach and the log-rank test.RESULTS In this study,we found that HCC patients with high expression of ASH1L possess a more recurrence rate as well as a decreased overall survival rate.ASH1L promotes the tumorigenicity of HCC and microRNA-142-3p exhibits reduced expression in HCC tissues and interacts with ASH1L through targeting the ASH1L 3′untranslated region.Furthermore,microRNA-142-3p promotes apoptosis and inhibits proliferation,invasion,and migration of HCC cell lines in vitro via ASH1L.For the exploration mechanism,we found ASH1L may promote an immunosuppressive microenvironment in HCC and ASH1L affects the expression of the cell junction protein zonula occludens-1,which is potentially relevant to the immune system.CONCLUSION Loss function of microRNA-142-3p induces cancer progression and immune evasion through upregulation of ASH1L in HCC.Both microRNA-142-3p and ASH1L can feature as new biomarker for HCC in the future.
