目的探讨miR-502对宫颈癌患者预后及其对宫颈癌Hela细胞增殖和转移的影响。方法通过数据库分析miR-502表达与宫颈癌患者临床病理特征的关系及其对预后的影响。在Hela细胞中过表达miR-502-3p后,采用CCK-8检测细胞增殖情况,Transwell实验...目的探讨miR-502对宫颈癌患者预后及其对宫颈癌Hela细胞增殖和转移的影响。方法通过数据库分析miR-502表达与宫颈癌患者临床病理特征的关系及其对预后的影响。在Hela细胞中过表达miR-502-3p后,采用CCK-8检测细胞增殖情况,Transwell实验检测细胞迁移和侵袭情况。结果数据库分析结果显示,高表达miR-502的宫颈癌患者总生存期更长,但与TNM分期无明显相关性。CCK-8检测结果显示,miR-502-3p类似物组细胞增殖率低于miR-502-3p抑制剂组[(78.82±1.41)% vs (124.06±7.72)%,P<0.05];Transwell实验检测结果显示,miR-502-3p类似物组细胞迁移率较miR-502-3p抑制剂组低[(85.39±7.19)% vs (121.17±8.20)%,P<0.05],细胞侵袭率亦较低[(85.32±7.12)% vs (117.13±7.37)%,P<0.05]。结论高表达miR-502的宫颈癌患者预后较好,可能与其抑制宫颈癌细胞增殖和转移有关。展开更多
Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's dis...Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.展开更多
文摘目的探讨miR-502对宫颈癌患者预后及其对宫颈癌Hela细胞增殖和转移的影响。方法通过数据库分析miR-502表达与宫颈癌患者临床病理特征的关系及其对预后的影响。在Hela细胞中过表达miR-502-3p后,采用CCK-8检测细胞增殖情况,Transwell实验检测细胞迁移和侵袭情况。结果数据库分析结果显示,高表达miR-502的宫颈癌患者总生存期更长,但与TNM分期无明显相关性。CCK-8检测结果显示,miR-502-3p类似物组细胞增殖率低于miR-502-3p抑制剂组[(78.82±1.41)% vs (124.06±7.72)%,P<0.05];Transwell实验检测结果显示,miR-502-3p类似物组细胞迁移率较miR-502-3p抑制剂组低[(85.39±7.19)% vs (121.17±8.20)%,P<0.05],细胞侵袭率亦较低[(85.32±7.12)% vs (117.13±7.37)%,P<0.05]。结论高表达miR-502的宫颈癌患者预后较好,可能与其抑制宫颈癌细胞增殖和转移有关。
基金supported by the National Institute on Aging (NIA)National Institutes of Health (NIH)+3 种基金Nos.K99AG065645,R00AG065645R00AG065645-04S1 (to SK)NIH research grants,NINDS,No.R01 NS115834NINDS/NIA,No.R01 NS115834-02S1 (to LG)。
文摘Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.
