目的探讨miR-185(Homo sapiens miR-185,miR-185)在结肠癌组织中的表达及其临床意义。方法运用原位杂交技术(hybridization in situ,ISH)检测miR-185在结肠癌及癌旁组织中的表达情况,分析其表达与结肠癌临床分期及淋巴结转移间的关系。...目的探讨miR-185(Homo sapiens miR-185,miR-185)在结肠癌组织中的表达及其临床意义。方法运用原位杂交技术(hybridization in situ,ISH)检测miR-185在结肠癌及癌旁组织中的表达情况,分析其表达与结肠癌临床分期及淋巴结转移间的关系。结果 ISH检测显示,在52例结肠癌组织中miR-185阳性表达率为13.46%,与癌旁对照组(84.62%)相比,差异有显著性(P<0.01);miR-185在结肠癌临床Ⅲ/Ⅳ期、Ⅰ/Ⅱ期和癌旁对照组中的光密度值分别为0.12±0.01、0.37±0.04、0.72±0.09,临床Ⅲ/Ⅳ期、Ⅰ/Ⅱ期分别与癌旁对照组比较,差异有显著性(P<0.05);淋巴结转移的与无淋巴结转移miR-185表达的光密度值分别为0.11±0.02、0.39±0.04,有淋巴结转移的miR-185表达水平低于无淋巴结转移,两者差异有显著性(P<0.01)。miR-185的表达随着结肠癌临床分期演进而降低结论miR-185与结肠癌的发生、临床进展、转移密切相关,其可能成为结肠癌治疗、预后判断的潜在生物学指标。展开更多
目的探讨微RNA(miRNA)-185-5p通过调控酪氨酸3/色氨酸5-单加氧酶激活蛋白(YWHAZ)对人非小细胞肺癌(NSCLC)细胞系A549增殖、侵袭凋亡和上皮间质转化的影响。方法获取GSE152702数据集,识别在NSCLC患者和健康者血液中差异性表达的miRNA,收...目的探讨微RNA(miRNA)-185-5p通过调控酪氨酸3/色氨酸5-单加氧酶激活蛋白(YWHAZ)对人非小细胞肺癌(NSCLC)细胞系A549增殖、侵袭凋亡和上皮间质转化的影响。方法获取GSE152702数据集,识别在NSCLC患者和健康者血液中差异性表达的miRNA,收集2018年7月至2019年9月在浙江大学附属第二医院确诊为NSCLC患者的癌组织及癌旁组织,使用RT-qPCR法检测miRNA-185-5p在NSCLC癌组织和细胞系中的表达情况。采用脂质体转染法,分别将miRNA-NC、miRNA-185-5p模拟物、miRNA-185-5p抑制剂和miRNA-185-5p模拟物+pcDNA3.0-YWHAZ转染至人NSCLC细胞A549中,Transwell实验检测各组A549细胞的侵袭能力,CCK-8实验检测各组A549细胞的增殖能力,脱氧核糖核酸末端转移酶介导的缺口末端标记(TUNEL)检测各组A549细胞的凋亡率,Western blot实验检测各组细胞中YWHAZ和上皮间质转化相关蛋白E-cadherin和Vimentin的表达情况。检索StarBase、Targetscan和Human protein Atlas数据库,韦恩图绘制与miRNA-185-5p含潜在互补结合位点及与NSCLC患者预后不良有关的基因。检索GEPIA数据库识别YWHAZ表达与肺鳞癌、肺腺癌患者预后的关系。生物学信息技术预测和双荧光素酶报告基因实验验证miRNA-185-5p和YWHAZ之间的关系。结果GSE152702数据集显示miRNA-185-5p在NSCLC患者中明显下调,miRNA-185-5p在NSCLC癌组织中的表达水平低于癌旁组织(P<0.01)。miRNA-185-5p在NSCLC细胞株中的表达水平低于正常人肺上皮细胞株(P<0.05),与miRNA-NC组比较,miRNA-185-5p模拟物组miRNA-185-5p表达水平明显升高(P<0.05),YWHAZ表达水平明显降低(P<0.01),miRNA-185-5p抑制剂组中miRNA-185-5p表达水平明显降低(P<0.05),YWHAZ表达水平明显升高(P<0.01)。过表达miRNA-185-5p能明显抑制A549细胞增殖、侵袭和上皮间质转化,并促进细胞凋亡(P<0.05)。GEPIA数据库显示,YWHAZ在肺鳞癌和肺腺癌患者中明显上调,且与患者预后不良有关。YWHAZ可与miRNA-185-5p靶向展开更多
Background:The liver plays a key role in regulating whole body cholesterol homeostasis.Hepatic cholesterol accumulation causes liver injury in fatty liver disease and hypercholesterolemia increases the risk of cardiov...Background:The liver plays a key role in regulating whole body cholesterol homeostasis.Hepatic cholesterol accumulation causes liver injury in fatty liver disease and hypercholesterolemia increases the risk of cardiovascular disease.MicroRNAs(miRNAs,miRs)have been shown to regulate various pathways in cholesterol metabolism.Recently,miR-185 has been shown to regulate sterol regulatory element-binding protein 2(SREBP2)and low-density lipoprotein receptor(LDLR)to modulate cholesterol syn-thesis and uptake.Materials and methods:The role of miR-185 in regulating diet-induced metabolic disorders were studied in liver-specific miRNA-185 knockout(L-miR-185 KO)mice.Results:L-miR-185 KO mice developed worsened hepatic steatosis upon high-fat high-cholesterol Western diet feeding with accumulation of triglyceride and cholesterol in the liver.In addition,L-miR-185 KO mice developed hypercholesterolemia upon Western diet feeding.Gene expression analysis showed that L-miR-185 KO mice did not show increased hepatic mRNA expression of SREBP2 or its targets LDLR and HMG-CoA reductase(HMGCR).Although expression of miR-185 mimic inhibited the mRNA of SREBP2,HMGCR and LDLR in HepG2 cells,miR-185 inhibitor did not increase the mRNA of SREBP2,HMGCR or LDLR in HepG2 cells.Conclusions:In conclusion,we reported that L-miR-185 KO mice were more sensitive to Western diet induced hepatic steatosis and hypercholesterolemia.The molecular mechanisms underlying these metabolic changes remain to be investigated in future studies.展开更多
文摘目的探讨微RNA(miRNA)-185-5p通过调控酪氨酸3/色氨酸5-单加氧酶激活蛋白(YWHAZ)对人非小细胞肺癌(NSCLC)细胞系A549增殖、侵袭凋亡和上皮间质转化的影响。方法获取GSE152702数据集,识别在NSCLC患者和健康者血液中差异性表达的miRNA,收集2018年7月至2019年9月在浙江大学附属第二医院确诊为NSCLC患者的癌组织及癌旁组织,使用RT-qPCR法检测miRNA-185-5p在NSCLC癌组织和细胞系中的表达情况。采用脂质体转染法,分别将miRNA-NC、miRNA-185-5p模拟物、miRNA-185-5p抑制剂和miRNA-185-5p模拟物+pcDNA3.0-YWHAZ转染至人NSCLC细胞A549中,Transwell实验检测各组A549细胞的侵袭能力,CCK-8实验检测各组A549细胞的增殖能力,脱氧核糖核酸末端转移酶介导的缺口末端标记(TUNEL)检测各组A549细胞的凋亡率,Western blot实验检测各组细胞中YWHAZ和上皮间质转化相关蛋白E-cadherin和Vimentin的表达情况。检索StarBase、Targetscan和Human protein Atlas数据库,韦恩图绘制与miRNA-185-5p含潜在互补结合位点及与NSCLC患者预后不良有关的基因。检索GEPIA数据库识别YWHAZ表达与肺鳞癌、肺腺癌患者预后的关系。生物学信息技术预测和双荧光素酶报告基因实验验证miRNA-185-5p和YWHAZ之间的关系。结果GSE152702数据集显示miRNA-185-5p在NSCLC患者中明显下调,miRNA-185-5p在NSCLC癌组织中的表达水平低于癌旁组织(P<0.01)。miRNA-185-5p在NSCLC细胞株中的表达水平低于正常人肺上皮细胞株(P<0.05),与miRNA-NC组比较,miRNA-185-5p模拟物组miRNA-185-5p表达水平明显升高(P<0.05),YWHAZ表达水平明显降低(P<0.01),miRNA-185-5p抑制剂组中miRNA-185-5p表达水平明显降低(P<0.05),YWHAZ表达水平明显升高(P<0.01)。过表达miRNA-185-5p能明显抑制A549细胞增殖、侵袭和上皮间质转化,并促进细胞凋亡(P<0.05)。GEPIA数据库显示,YWHAZ在肺鳞癌和肺腺癌患者中明显上调,且与患者预后不良有关。YWHAZ可与miRNA-185-5p靶向
基金This study was supported by the USA National Institutes of Health(NIH)grant 1R01 DK117965-01A1(T.Li).
文摘Background:The liver plays a key role in regulating whole body cholesterol homeostasis.Hepatic cholesterol accumulation causes liver injury in fatty liver disease and hypercholesterolemia increases the risk of cardiovascular disease.MicroRNAs(miRNAs,miRs)have been shown to regulate various pathways in cholesterol metabolism.Recently,miR-185 has been shown to regulate sterol regulatory element-binding protein 2(SREBP2)and low-density lipoprotein receptor(LDLR)to modulate cholesterol syn-thesis and uptake.Materials and methods:The role of miR-185 in regulating diet-induced metabolic disorders were studied in liver-specific miRNA-185 knockout(L-miR-185 KO)mice.Results:L-miR-185 KO mice developed worsened hepatic steatosis upon high-fat high-cholesterol Western diet feeding with accumulation of triglyceride and cholesterol in the liver.In addition,L-miR-185 KO mice developed hypercholesterolemia upon Western diet feeding.Gene expression analysis showed that L-miR-185 KO mice did not show increased hepatic mRNA expression of SREBP2 or its targets LDLR and HMG-CoA reductase(HMGCR).Although expression of miR-185 mimic inhibited the mRNA of SREBP2,HMGCR and LDLR in HepG2 cells,miR-185 inhibitor did not increase the mRNA of SREBP2,HMGCR or LDLR in HepG2 cells.Conclusions:In conclusion,we reported that L-miR-185 KO mice were more sensitive to Western diet induced hepatic steatosis and hypercholesterolemia.The molecular mechanisms underlying these metabolic changes remain to be investigated in future studies.
