Analyse The Clinical Risk Factors Of Hepatocellular Carcinoma Based On Cirrhosis And Study On The Microrna Expression Profiles Of Liver Cancer And Cirrhosis

Objective:1) To survey the genetics characteristics of liver cirrhosis and risk factors of HCC incident of liver cirrhosis in XinJiang province. and analyze biochemical characteristics of liver cirrhosis. See if there were something different when there are HBV infection mixed with alcoholic.2) Screening on the microRNA expression profiles in liver cancer and cirrhosis with HBV.3) To detect the expression of the most differential miRNA in patients with hepatocellular carcinoma. To provide new ideas and ways for the study of the molecular mechanism of primary hepatocellular carcinoma and to plays foundation for exploring biomolecular marker of HCC to early diagnosis. Methods:1) All of liver cirrhosis patients who were hospitalized in the First affiliated hospital of XinJiang Medical University in past11years were included, they were differed into different groups according to the genetics of liver cirrhosis, hypothesis mild alcohol intake、HBV、HCV、alcohol、gender、age、ethnicity、diabetes mellitus2as the HCC risk factors; liver function test, lipoprotein and regular blood test results from patients’ data were compared among HBV combined mild alcohol intake liver cirrhosis, HBV related liver cirrhosis and alcoholic cirrhosis. Data were analyzed by SPSS13.0software. Using co-variant analysis method.2) In1169patients with HCC from the7212patients, to select9patients treated with resection of hepatocellular carcinoma, take the HCC specimen of the operation (experimental group) and more than5cm from the tumor of liver cirrhosis specimens (control group). Extraction of the total RNA in hepatocellular carcinoma tissues and paired liver cirrhosis tissues by Trizol method, to quantitative analysis by the spectrophotometer NanoDropND-1000, and extracted total RNA denaturing by gel electrophoresis. To microRNA chip hybridization by the microarray technique, the results to scanning reading, Finally, Significance Analysis of Microarrays software (SAM, version2.1) selected differentially expressed genes. To validate2miRNAs (hsa-miR-1269and hsa-miR-199b-3p) by the Real-time fluorescence quantitative PCR.(3).To detect the expression level of hsa-miR-1269in hepatocellular carcinoma and cirrhosis tissues by Real-time PCR. Results:1)7212liver cirrhosis patients were included, the most common fifth genetics of liver cirrhosis in Xiniang were HBV55.10%, HCV13.42%, PBC9.28%, alcoholic cirrhosis6.36%, cryptogenic cirrhosis9.01%; HCC16.5%, death rate6.3%during in hospital,12.1%patients combined with diabetes mellitus.There were3976liver cirrhosis due to HBV infection,24.25%of them were HBV combined with mild alcoholic intake. When HBV infection mixed with mild alcoholic intake, highest level of white cell count, hemoglobin, mean corpuscular volume, y-glutamyltranspeptidase and uric acid were observed (P<0.001) comparing with alcoholic cirrhosis and HBV related liver cirrhosis; HBV. HCV. alcoholic factor. male、aged were risk factors of HCC incident by multivariate analysis.2) Differential expression of miRNAs in HCC and cirrhosis tissues:The differentially expressed miRNAs of the experimental group and the control group has15, respectively, hsa-miR-1269, hsa-miR-18a, hsa-miR-1180, hsa-miR-1301, hsa-miR-182, hsa-miR-222, hsa-miR-362-5p, hsa-miR-99a, hsa-miR-lOa, hsa-miR-30a, hsa-miR-30e, hsa-miR-139-5p, hsa-miR-422a, hsa-miR-199a-3p, hsa-miR-199b-3p. Real time fluorescence quantitative PCR results:the relative abundance of hsa-miR-1269in the experimental group and the control group in abundance were12.058±3.954and16.678±2.352(P=0.0096). The relative abundance of hsa-miR-199b-3p expression in the experimental group and the control group in abundance were5.417±1.633and3.033±0.768(P=0.0028). The up-regulation of hsa-miR-1269expression and the dow-regulation expression of hsa-miR-199b-3p in hepatocellular carcinoma, miRNAs microarray and Real time PCR were consistent.3) Expression of hsa-miR-1269in HCC tissues:Real time fluorescence quantitative PCR results:the average abundance of hsa-miR-1269in the experimental group and the control group were12.634±4.004and16.963±1.832(P=0.0035). Conclusion:1) HBV is the dominate genetic for liver cirrhosis in XinJiang. But HBV mixed with alcoholic may be the genetical characteristic of the area. HBV combined with mild alcoholic liver cirrhosis endure the higher oxidative stress comparing with HBV-related liver cirrhosis, hint indirectly that even mild alcohol intake may increase liver cirrhosis developed in HBV infection population; HBV、HCV、alcoholic、male、aged were HCC risk factors. diabetes mellitus2and ethnicity (Uygur) didinot increase HCC incident comparing with HBV related liver cirrhosis.2) The study found15different miRNAs in HCC and liver cirrhosis tissues with HBV, which hsa-miR-1180, hsa-miR-1301has not been reported in the literatures, and hsa-miR-1269and hsa-miR-10a related to the occurrence and development of other tumors, but not in HCC.3) To all15miRNAs, hsa-miR-1269expression was significantly higher in HCC tissue, so, it may be a functionally similar oncogenic miRNAs.