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
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
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。
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.
