Expression Analysis Of Mrnas And Mirnas In Tissue And Serum Of Hepatocellular Carcinoma Patients

Part OneObjective:To identify differentially expressed mRNAs between hepatocellular carcinoma tissues and its matched adjacent normal liver tissues, and to carry out the bioinformatics analysis.Methods:Agilent 8x60K microarray technology was used to detect the changes of mRNA expression between hepatocellular carcinoma tissues and its matched adjacent normal liver tissues. To filter differentially expressed mRNAs, we did GO and Pathway analysis with the help of bioinformatics methods. Then Real-time PCR was applied to verify microarray data.Results:924 mRNAs exhibited higher expression in the hepatocellular carcinoma tissues than its matched adjacent normal liver tissues. In comparison with the adjacent normal tissues, the hepatocellular carcinoma tissues showed down-regulted expressions of 1770 mRNAs. GO and Pathway analysis involved in transcription process, REDOX, signal transduction ion transport, immune response, cell adhesion, and binding functions. The results of Real-time PCR were in high concordance with microarray results.Conclusion:Differentially expressed mRNAs may include signal transduction, immune response and many other key links, and may provide new ideas for early diagnosis and target therapy of hepatocellular carcinoma.Part TwoObjective:To identify differentially expressed miRNAs between hepatocellular carcinoma tissues and its matched adjacent normal liver tissues, and to predict the target genes of selected miRNAs.Methods:Microarray technology was used to detect the changes of miRNA expression between hepatocellular carcinoma tissues and its matched adjacent normal liver tissues. We predict the target genes of selected miRNAs by bioinformatics tools. Then Real-time PCR was applied to verify microarray data.Results:21 miRNAs exhibited higher expression in the hepatocellular carcinoma tissues than its matched adjacent normal liver tissues. In comparison with the adjacent normal tissues, the hepatocellular carcinoma tissues showed down-regulted expressions of 12 miRNAs. We obtained some valuable miRNAs and its target genes by comparing the results of mRNA microarray in part one with the results of target gene prediction. The results of Real-time PCR were in high concordance with microarray results.Conclusion:The selected differentially expressed miRNAs and its specific target genes may be active in the development of hepatocellular carcinoma, and these genes may create a new path for the specific targeted therapy of hepatocellular carcinoma patients.Part ThreeObjective:To detect expression levels of selected miRNAs in serum of hepatocellular carcinoma patients, and to carry out clinical analysis.Methods:We detected the expression levels of selected miRNAs (miR-155, miR-96, miR-99a) in serum of hepatocellular carcinoma patients, then compared with the expression levels in tissues of hepatocellular carcinoma patients, and carried out clinical analysis.Results:The results of selected miRNAs (miR-155, miR-96, miR-99a) in serum of hepatocellular carcinoma patients were in high concordance with tissues. These differentially expressed miRNAs had no obvious relation with age, sex, tumor number, tumor size, AFP levels and cirrhosis (P>0.05), but had obvious relation with TNM stages, differentiation and metastasis (P<0.05)Conclusion:Some miRNAs expressed differentially both in tissues and in serum of hepatocellular carcinoma patients. These differentially expressed miRNAs may be related with TNM stages, differentiation and metastasis and these differentially expressed miRNAs may be as a new biological indicator of hepatocellular carcinoma.