This study aimed to investigate the correlation between serum miR-154-5p and urinary albumin to creatinine ratio(UACR)in patients with type 2 diabetes mellitus(T2DM)and the association with biomarkers of inflammation ...This study aimed to investigate the correlation between serum miR-154-5p and urinary albumin to creatinine ratio(UACR)in patients with type 2 diabetes mellitus(T2DM)and the association with biomarkers of inflammation and fibrosis in diabetic kidney disease(DKD).A total of 390 patients with T2DM were divided into three groups:normal albuminuria(UACR<30 mg/g,n=136,NA),microalbuminuria(UACR at 30-300 mg/g,n=132,MA),and clinical albuminuria(UACR>300 mg/g,n=122,CA).Circulating miR-154-5p,inflammatory(C-reactive protein(CRP);erythrocyte sedimentation rate(ESR);and tumor necrosis factor-a(TNF-α)and fibrotic markers(vascular endothelial growth factor(VEGF);transforming growth factor-β1(TGF-β1);and fibronectin(FN),and other biochemical indicators were assessed via real-time PCR,enzyme-linked immunosorbent assay,and chemiluminescence assay in patients with T2DM and 138 control subjects(NC).UACR,miR-154-5p,glycated hemoglobin(HbA1c),serum creatinine(sCr),blood urea nitrogen(BUN),ESR,CRP,VEGF,TNF-α,TGF-β1,and FN were significantly higher and the estimated glomerular filtration rate(eGFR)was significantly lower in NA,MA,and CA groups than in NC subjects(P<0.05).Elevated levels of UACR and miR-154-5p were directly correlated with HbA1c,sCr,BUN,ESR,CRP,VEGF,TNF-α,TGF-β1,and FN and negatively correlated with eGFR(P<0.05).miR-154-5p,HbA1c,sCr,BUN,eGFR,ESR,CRP,VEGF,TNF-α,TGF-β1,and FN were important factors affecting UACR.These findings indicated that elevated serum miR-154-5p is significantly correlated with high UACR in patients with T2DM and may offer a novel reference for the early diagnosis of DKD.展开更多
探讨乙型肝炎病毒(Hepatitis B virus,HBV)通过miR⁃154⁃5p/STAT3轴促进肝癌细胞增殖和迁移的影响。用含有1.3倍超长HBV基因组的腺病毒感染HepG2细胞,以正常HepG2作为对照(NC),记为HBV组和NC组。采用实时荧光定量PCR(qRT⁃PCR)检测HBV组...探讨乙型肝炎病毒(Hepatitis B virus,HBV)通过miR⁃154⁃5p/STAT3轴促进肝癌细胞增殖和迁移的影响。用含有1.3倍超长HBV基因组的腺病毒感染HepG2细胞,以正常HepG2作为对照(NC),记为HBV组和NC组。采用实时荧光定量PCR(qRT⁃PCR)检测HBV组、NC组、稳定表达HBV的人肝癌细胞HepG2.2.15及人肝癌细胞HepG2中miR⁃154⁃5p表达水平。按照脂质体法将miR⁃NC模拟物、过表达miR⁃154⁃5p、过表达miR⁃154⁃5p+空载体pcDNA、过表达miR⁃154⁃5p+过表达STAT3转染至HepG2.2.15细胞中,记为miR⁃NC组、miR⁃154⁃5p组、miR⁃154⁃5p+pcDNA组、miR⁃154⁃5p+STAT3组。细胞计数试剂盒(CCK8)、克隆形成实验检测细胞增殖;伤口愈合实验检测细胞迁移;蛋白免疫印迹(Western blot)法检测信号转导与转录因子3(STAT3)、增殖细胞核抗原(PCNA)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)蛋白表达。荧光素酶实验检测miR⁃154⁃5p和STAT3的关系。与NC组相比,HBV组内miR⁃154⁃5p表达水平降低;与HepG2细胞相比,HepG2.2.15细胞中miR⁃154⁃5p表达水平明显降低。上调miR⁃154⁃5p明显降低细胞活性、克隆形成数、迁移率,并抑制了PCNA、MMP2和MMP9蛋白表达。miR⁃154⁃5p靶向负调控STAT3的表达,且过表达STAT3可以逆转上调miR⁃154⁃5p对HepG2.2.15细胞增殖、迁移的抑制作用。乙型肝炎病毒通过降低miR⁃154⁃5p表达上调STAT3,进而促进肝癌细胞的增殖和迁移。展开更多
Constitutive hedgehog （Hh） signaling is associated with the genesis of medulloblastomas （MB）. The objective of this study is to identify special microRNAs （miRNAs） regulated by the Hh pathway, and to clarify the...Constitutive hedgehog （Hh） signaling is associated with the genesis of medulloblastomas （MB）. The objective of this study is to identify special microRNAs （miRNAs） regulated by the Hh pathway, and to clarify the role of miRNAs during the genesis of MB induced by sustained Hh activation. In the primary screening, we used stemloop RT-PCR to test the expression of 90 different miRNAs in the wildtype （WT） and Ptc-/- MEF cell lines. In the secondary screening, the miRNAs screened from the first screening were validated in the Sufu-/- MEF cell lines. We then verified the expression of miRNAs both in the normal cerebellar tissues and the MB induced by activated Hh pathway, and examined the expression of the other 21 miRNA members of the miR-154 cluster in the MB and normal cerebellum. In the first screening, 13 miRNAs showed significant differential expression in WT and Ptc-/- MEF cell lines, while 10 of them had significant difference in the Sufu-/- MEF cell line. Compared to the normal mouse cerebellum, only 2 miRNAs in 15 miRNAs were differentially expressed between the MB and normal cerebellar tissues. Among 21 members of the miR-154 cluster, 6 miRNAs were downregulated in the MB. Our study demonstrated that miR-154 may be regulated by the Hh pathway, and the activation of the Hh pathway led to the downregulation of the miR-154 cluster, resulting in the genesis of MB.展开更多
基金This study was supported by the Higher School“High-end Talent Team Construction”of Liaoning Province(No.[2014]187)the Natural Science Foundation of Liaoning Province(No.201602862),Liaoning Province,China.
文摘This study aimed to investigate the correlation between serum miR-154-5p and urinary albumin to creatinine ratio(UACR)in patients with type 2 diabetes mellitus(T2DM)and the association with biomarkers of inflammation and fibrosis in diabetic kidney disease(DKD).A total of 390 patients with T2DM were divided into three groups:normal albuminuria(UACR<30 mg/g,n=136,NA),microalbuminuria(UACR at 30-300 mg/g,n=132,MA),and clinical albuminuria(UACR>300 mg/g,n=122,CA).Circulating miR-154-5p,inflammatory(C-reactive protein(CRP);erythrocyte sedimentation rate(ESR);and tumor necrosis factor-a(TNF-α)and fibrotic markers(vascular endothelial growth factor(VEGF);transforming growth factor-β1(TGF-β1);and fibronectin(FN),and other biochemical indicators were assessed via real-time PCR,enzyme-linked immunosorbent assay,and chemiluminescence assay in patients with T2DM and 138 control subjects(NC).UACR,miR-154-5p,glycated hemoglobin(HbA1c),serum creatinine(sCr),blood urea nitrogen(BUN),ESR,CRP,VEGF,TNF-α,TGF-β1,and FN were significantly higher and the estimated glomerular filtration rate(eGFR)was significantly lower in NA,MA,and CA groups than in NC subjects(P<0.05).Elevated levels of UACR and miR-154-5p were directly correlated with HbA1c,sCr,BUN,ESR,CRP,VEGF,TNF-α,TGF-β1,and FN and negatively correlated with eGFR(P<0.05).miR-154-5p,HbA1c,sCr,BUN,eGFR,ESR,CRP,VEGF,TNF-α,TGF-β1,and FN were important factors affecting UACR.These findings indicated that elevated serum miR-154-5p is significantly correlated with high UACR in patients with T2DM and may offer a novel reference for the early diagnosis of DKD.
文摘探讨乙型肝炎病毒(Hepatitis B virus,HBV)通过miR⁃154⁃5p/STAT3轴促进肝癌细胞增殖和迁移的影响。用含有1.3倍超长HBV基因组的腺病毒感染HepG2细胞,以正常HepG2作为对照(NC),记为HBV组和NC组。采用实时荧光定量PCR(qRT⁃PCR)检测HBV组、NC组、稳定表达HBV的人肝癌细胞HepG2.2.15及人肝癌细胞HepG2中miR⁃154⁃5p表达水平。按照脂质体法将miR⁃NC模拟物、过表达miR⁃154⁃5p、过表达miR⁃154⁃5p+空载体pcDNA、过表达miR⁃154⁃5p+过表达STAT3转染至HepG2.2.15细胞中,记为miR⁃NC组、miR⁃154⁃5p组、miR⁃154⁃5p+pcDNA组、miR⁃154⁃5p+STAT3组。细胞计数试剂盒(CCK8)、克隆形成实验检测细胞增殖;伤口愈合实验检测细胞迁移;蛋白免疫印迹(Western blot)法检测信号转导与转录因子3(STAT3)、增殖细胞核抗原(PCNA)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)蛋白表达。荧光素酶实验检测miR⁃154⁃5p和STAT3的关系。与NC组相比,HBV组内miR⁃154⁃5p表达水平降低;与HepG2细胞相比,HepG2.2.15细胞中miR⁃154⁃5p表达水平明显降低。上调miR⁃154⁃5p明显降低细胞活性、克隆形成数、迁移率,并抑制了PCNA、MMP2和MMP9蛋白表达。miR⁃154⁃5p靶向负调控STAT3的表达,且过表达STAT3可以逆转上调miR⁃154⁃5p对HepG2.2.15细胞增殖、迁移的抑制作用。乙型肝炎病毒通过降低miR⁃154⁃5p表达上调STAT3,进而促进肝癌细胞的增殖和迁移。
文摘Constitutive hedgehog （Hh） signaling is associated with the genesis of medulloblastomas （MB）. The objective of this study is to identify special microRNAs （miRNAs） regulated by the Hh pathway, and to clarify the role of miRNAs during the genesis of MB induced by sustained Hh activation. In the primary screening, we used stemloop RT-PCR to test the expression of 90 different miRNAs in the wildtype （WT） and Ptc-/- MEF cell lines. In the secondary screening, the miRNAs screened from the first screening were validated in the Sufu-/- MEF cell lines. We then verified the expression of miRNAs both in the normal cerebellar tissues and the MB induced by activated Hh pathway, and examined the expression of the other 21 miRNA members of the miR-154 cluster in the MB and normal cerebellum. In the first screening, 13 miRNAs showed significant differential expression in WT and Ptc-/- MEF cell lines, while 10 of them had significant difference in the Sufu-/- MEF cell line. Compared to the normal mouse cerebellum, only 2 miRNAs in 15 miRNAs were differentially expressed between the MB and normal cerebellar tissues. Among 21 members of the miR-154 cluster, 6 miRNAs were downregulated in the MB. Our study demonstrated that miR-154 may be regulated by the Hh pathway, and the activation of the Hh pathway led to the downregulation of the miR-154 cluster, resulting in the genesis of MB.
