As the most abundant liver-specific microRNA,mi-croRNA-122(miR-122)is involved in various physio-logical processes in hepatic function as well as in liver pathology.There is now compelling evidence that miR-122,as a r...As the most abundant liver-specific microRNA,mi-croRNA-122(miR-122)is involved in various physio-logical processes in hepatic function as well as in liver pathology.There is now compelling evidence that miR-122,as a regulator of gene networks and pathways in hepatocytes,plays a central role in diverse aspects of hepatic function and in the progress of liver diseases.This liver-enriched transcription factors-regulated miRNA promotes differentiation of hepatocytes and regulates lipid metabolism.With regard to liver diseases,miR-122 was shown to stimulate hepatitis C virus(HCV)replication through a unique and unusual interaction with two binding sites in the 5′-UTR of HCV genome to mediate the stability of the viral RNA,whereas inhibit the expression and replication of hepatitis B virus(HBV)by a miR-122-cylin G1/p53-HBV enhancer regulatory pathway.In addition,miR-122 acts as a suppressor of cell prolif-eration and malignant transformation of hepatocytes with remarkable tumor inhibition activity.Notably,a clinical trial targeting miR-122 with the anti-miR-122 oli-gonucleotides miravirsen,the first miRNA targeted drug,has been initiated for treatment of HCV infection.With further understanding of the comprehensive roles of miR-122 in hepatic functions and the mechanisms in-volved in miR-122 down-regulation in chronic hepatitis or hepatocellular carcinoma,miR-122 appears to be a promising candidate for effective therapeutic ap-proaches against tumor and infectious diseases.展开更多
MiR-122 is one of the non-coding RNAs which showed its effects on the lipo-metablism, virus infection and HCC forming through regulation of liver gene expression. Its eukaryotic expression vector was constructed by us...MiR-122 is one of the non-coding RNAs which showed its effects on the lipo-metablism, virus infection and HCC forming through regulation of liver gene expression. Its eukaryotic expression vector was constructed by using pSuper which was widely applied in the siRNA expression. The precursor of human miR-122 gene was amplified by polymerase chain reaction （PCR） from the human genomic DNA. The positive clones were screened by PCR and restriction enzyme digestion. The new expression vector of miR-122 was named pHsa-m122. PHsa-m122 and its controls were transfected to HepG2 cells. The miR-122 expression activity was evaluated by GFP122i sensor reporter plasmid through fluorescence detection and Western blot. It was shown that the fluorescence intensity of GFP122si and pHsa-m122 co-transfection group was weaker than that of the controls, so the functional activity of expressed miR-122 was detected. When HepG2 cells were co-transfected with HBV1.3 and pHsa-m122 plasmids, the results showed miR-122 may down-regulate the gene expression of HBV. The human liver specific microRNA eukaryotic expression vector of miR-122 was constructed successfully, which may facilitate further study of its function in the development of liver virus infection diseases and HCC. Cellular ＆ Molecular Immunology.展开更多
Hepatocellular carcinoma(HCC)is one of the common lethal types of tumor all over the world.The lethality of HCC accounts for many reasons.One of them,the lack of reliable diagnostic markers at the early stage,in this ...Hepatocellular carcinoma(HCC)is one of the common lethal types of tumor all over the world.The lethality of HCC accounts for many reasons.One of them,the lack of reliable diagnostic markers at the early stage,in this context,serum miRNAs became promising diagnostic biomarkers.Herein,we aimed to identify the predictive value of two miRNAs(miR-122 and miR-224)in plasma of patients with HCC preceded by chronic HCV infection.Taqman miRNA assays specific for hsa-miR-122 and hsa-miR-224 were used to assess the expression levels of the chosen miRNAs in plasma samples collected from three groups;40 patients with HCC related to HCV,40 with CHC patients and 20 healthy volunteers.This study revealed that the mean plasma values of miRNA-122 were significantly lower among HCC group when compared to CHC and control groups(P<0.001).Whereas,miR-224 mean plasma values were significantly higher among HCC group when compared to both CHC group and control group.Moreover,it was found that miR-122 can predict development of HCC at cut-off value<0.67(RQ)and(AUC Z 0.98,P<0.001).As regards miR-224,it can predict development of HCC at cut-off value>1.2(RQ)and(AUC Z 0.93,P<0.001),while the accuracy of AFP to diagnose HCC was(AUC:0.619;P Z 0.06).In conclusion,the expression plasma of miR-122 and miR-224 could be used as noninvasive biomarkers for the early prediction of developing HCC at the early stage.展开更多
MicroRNAs （miRNAs） are endogenous small non-coding RNAs that repress their targets at post transcriptional level.Existing studies have shown that miRNAs are important regulatory genes in hepatocellular carcinoma （...MicroRNAs （miRNAs） are endogenous small non-coding RNAs that repress their targets at post transcriptional level.Existing studies have shown that miRNAs are important regulatory genes in hepatocellular carcinoma （HCC）,as either tumor suppressors or oncogenes.MiR-122 is normally downregulated in HCC and regarded as a tumor suppressor.Recently miR-122 has been reported to be regulated by CEBPA,which is then involved in a novel pathway to influence proliferation of tumor cells.However it is unknown whether CEBPA is regulated by miRNAs in HCC.In this study,we find that miR182 is upregulated in HCC model rat,and represses CEBPA in both rat and human.This further improves the current CEBPA/miR-122 pathway that controls the proliferation of tumor cells.These results suggest that miR-182 is a potential oncogene in HCC and could be used as a diagnostic marker and drug target of HCC.展开更多
AIM: To investigate whether expression of selected mi RNAs obtained from fibrotic liver biopsies correlate with fibrosis stage.METHODS: Altogether, 52 patients were enrolled in the study representing various etiologic...AIM: To investigate whether expression of selected mi RNAs obtained from fibrotic liver biopsies correlate with fibrosis stage.METHODS: Altogether, 52 patients were enrolled in the study representing various etiologic backgrounds of fibrosis: 24 cases with chronic hepatitis infections(types B, C), 19 with autoimmune liver diseases(autoimmune hepatitis, primary biliary cirrhosis, primary sclerosing cholangitis, overlapping syndrome cases), and 9 of mixed etiology(alcoholic and nonalcoholic steatosis, cryptogenic cases). Severity of fibrosis was determined by both histologic staging using the METAVIR scoring system and noninvasive transient elastography. Following RNAisolation, expression levels of mi R-21, mi R-122, mi R-214, mi R-221, mi R-222, and mi R-224 were determined using Taq Man Micro RNA Assays applying mi R-140 as the reference. Selection of mi RNAs was based on their characteristic up- or downregulation observed in hepatocellular carcinoma. Relative expression of mi RNAs was correlated with fibrosis stage and liver stiffness(LS) value measured by transient elastography, as well as with serum alanine aminotransferase(ALT) level.RESULTS: The expression of individual mi RNAs showed deregulated patterns in stages F1-F4 as compared with stage F0, but only the reduced level of mi R-122 in stage F4 was statistically significant(P < 0.04). When analyzing mi RNA expression in relation to fibrosis, levels of mi R-122 and mi R-221 showed negative correlations with fibrosis stage, and mi R-122 was found to correlate negatively and mi R-224 positively with LS values(all P < 0.05). ALT levels displayed a positive correlation with mi R-21(P < 0.04). Negative correlations were observed in the fibrosis samples of mixed etiology between mi R-122 and fibrosis stage and LS values(P < 0.05), and in the samples of chronic viral hepatitis, between mi R-221 and fibrosis stage(P < 0.01), whereas mi R-21 showed positive correlation with ALT values in the samples of autoimmune liver diseases(P < 0.03). The results also revealed 展开更多
基金supported by grants from the National Natural Science Foundation of China(Grant Nos.30970146,91029724,and 81021003).
文摘As the most abundant liver-specific microRNA,mi-croRNA-122(miR-122)is involved in various physio-logical processes in hepatic function as well as in liver pathology.There is now compelling evidence that miR-122,as a regulator of gene networks and pathways in hepatocytes,plays a central role in diverse aspects of hepatic function and in the progress of liver diseases.This liver-enriched transcription factors-regulated miRNA promotes differentiation of hepatocytes and regulates lipid metabolism.With regard to liver diseases,miR-122 was shown to stimulate hepatitis C virus(HCV)replication through a unique and unusual interaction with two binding sites in the 5′-UTR of HCV genome to mediate the stability of the viral RNA,whereas inhibit the expression and replication of hepatitis B virus(HBV)by a miR-122-cylin G1/p53-HBV enhancer regulatory pathway.In addition,miR-122 acts as a suppressor of cell prolif-eration and malignant transformation of hepatocytes with remarkable tumor inhibition activity.Notably,a clinical trial targeting miR-122 with the anti-miR-122 oli-gonucleotides miravirsen,the first miRNA targeted drug,has been initiated for treatment of HCV infection.With further understanding of the comprehensive roles of miR-122 in hepatic functions and the mechanisms in-volved in miR-122 down-regulation in chronic hepatitis or hepatocellular carcinoma,miR-122 appears to be a promising candidate for effective therapeutic ap-proaches against tumor and infectious diseases.
基金financially supported by the grants from Natural Science Foundation of Hubei Province(No.2006ABA143)Foundation of Tongji Medical College of Huazhong University of Science and Technology(No.01510747).
文摘MiR-122 is one of the non-coding RNAs which showed its effects on the lipo-metablism, virus infection and HCC forming through regulation of liver gene expression. Its eukaryotic expression vector was constructed by using pSuper which was widely applied in the siRNA expression. The precursor of human miR-122 gene was amplified by polymerase chain reaction （PCR） from the human genomic DNA. The positive clones were screened by PCR and restriction enzyme digestion. The new expression vector of miR-122 was named pHsa-m122. PHsa-m122 and its controls were transfected to HepG2 cells. The miR-122 expression activity was evaluated by GFP122i sensor reporter plasmid through fluorescence detection and Western blot. It was shown that the fluorescence intensity of GFP122si and pHsa-m122 co-transfection group was weaker than that of the controls, so the functional activity of expressed miR-122 was detected. When HepG2 cells were co-transfected with HBV1.3 and pHsa-m122 plasmids, the results showed miR-122 may down-regulate the gene expression of HBV. The human liver specific microRNA eukaryotic expression vector of miR-122 was constructed successfully, which may facilitate further study of its function in the development of liver virus infection diseases and HCC. Cellular ＆ Molecular Immunology.
文摘Hepatocellular carcinoma(HCC)is one of the common lethal types of tumor all over the world.The lethality of HCC accounts for many reasons.One of them,the lack of reliable diagnostic markers at the early stage,in this context,serum miRNAs became promising diagnostic biomarkers.Herein,we aimed to identify the predictive value of two miRNAs(miR-122 and miR-224)in plasma of patients with HCC preceded by chronic HCV infection.Taqman miRNA assays specific for hsa-miR-122 and hsa-miR-224 were used to assess the expression levels of the chosen miRNAs in plasma samples collected from three groups;40 patients with HCC related to HCV,40 with CHC patients and 20 healthy volunteers.This study revealed that the mean plasma values of miRNA-122 were significantly lower among HCC group when compared to CHC and control groups(P<0.001).Whereas,miR-224 mean plasma values were significantly higher among HCC group when compared to both CHC group and control group.Moreover,it was found that miR-122 can predict development of HCC at cut-off value<0.67(RQ)and(AUC Z 0.98,P<0.001).As regards miR-224,it can predict development of HCC at cut-off value>1.2(RQ)and(AUC Z 0.93,P<0.001),while the accuracy of AFP to diagnose HCC was(AUC:0.619;P Z 0.06).In conclusion,the expression plasma of miR-122 and miR-224 could be used as noninvasive biomarkers for the early prediction of developing HCC at the early stage.
基金supported in part by a start-up grant of Kunming University of Science and Technology given to Yun Zhenga major projects of Shanghai Municipal Health Bureau(No.20100222) given to Xiaolin Wanga Youth Fund of Zhongshan Hospital,Fudan University(No.201102) given to Chenggang Wang
文摘MicroRNAs （miRNAs） are endogenous small non-coding RNAs that repress their targets at post transcriptional level.Existing studies have shown that miRNAs are important regulatory genes in hepatocellular carcinoma （HCC）,as either tumor suppressors or oncogenes.MiR-122 is normally downregulated in HCC and regarded as a tumor suppressor.Recently miR-122 has been reported to be regulated by CEBPA,which is then involved in a novel pathway to influence proliferation of tumor cells.However it is unknown whether CEBPA is regulated by miRNAs in HCC.In this study,we find that miR182 is upregulated in HCC model rat,and represses CEBPA in both rat and human.This further improves the current CEBPA/miR-122 pathway that controls the proliferation of tumor cells.These results suggest that miR-182 is a potential oncogene in HCC and could be used as a diagnostic marker and drug target of HCC.
基金Supported by Grant from the National Scientific Research Fund,OTKA K101435 and K108548
文摘AIM: To investigate whether expression of selected mi RNAs obtained from fibrotic liver biopsies correlate with fibrosis stage.METHODS: Altogether, 52 patients were enrolled in the study representing various etiologic backgrounds of fibrosis: 24 cases with chronic hepatitis infections(types B, C), 19 with autoimmune liver diseases(autoimmune hepatitis, primary biliary cirrhosis, primary sclerosing cholangitis, overlapping syndrome cases), and 9 of mixed etiology(alcoholic and nonalcoholic steatosis, cryptogenic cases). Severity of fibrosis was determined by both histologic staging using the METAVIR scoring system and noninvasive transient elastography. Following RNAisolation, expression levels of mi R-21, mi R-122, mi R-214, mi R-221, mi R-222, and mi R-224 were determined using Taq Man Micro RNA Assays applying mi R-140 as the reference. Selection of mi RNAs was based on their characteristic up- or downregulation observed in hepatocellular carcinoma. Relative expression of mi RNAs was correlated with fibrosis stage and liver stiffness(LS) value measured by transient elastography, as well as with serum alanine aminotransferase(ALT) level.RESULTS: The expression of individual mi RNAs showed deregulated patterns in stages F1-F4 as compared with stage F0, but only the reduced level of mi R-122 in stage F4 was statistically significant(P < 0.04). When analyzing mi RNA expression in relation to fibrosis, levels of mi R-122 and mi R-221 showed negative correlations with fibrosis stage, and mi R-122 was found to correlate negatively and mi R-224 positively with LS values(all P < 0.05). ALT levels displayed a positive correlation with mi R-21(P < 0.04). Negative correlations were observed in the fibrosis samples of mixed etiology between mi R-122 and fibrosis stage and LS values(P < 0.05), and in the samples of chronic viral hepatitis, between mi R-221 and fibrosis stage(P < 0.01), whereas mi R-21 showed positive correlation with ALT values in the samples of autoimmune liver diseases(P < 0.03). The results also revealed
