Development of super-low-invasive liver-biopsy-method using molecular genetics analysis

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1. Evaluation of liver damage using molecular genetics methodWe analyzed gene expression profile of non-cancer lesion of 121 patients using oligo array. We finally obtained samples from 81 patients who could be analyzed with liver mean fibrous index (MFI) using Azan staining.2. Comparing patients in hepatitis C and hepatitis BsAmong patients with hepatitis C, genes related Interferon or immune function were observed. GIP2 or IFI27, which located at the top of the list, were observed in upregulated expression area in cancer area of HCV-positive HCC patients. This gene list was identified to be different expression pattern of the patients with Hepatitis B and C. Lots of genes were classified in type C and others.3. The evaluation of result using analysis with super-thin-needle methodWe obtained the amount of total RNA using 18G:30.4±4.4μg and 20G:15.3±1.4μg. We confirm that the cluster using the same patients was located at the same position. Furthermore, we obtained the amount of total RNA using more-thin needle of 22G:4.9±0.9μg、23G:3.4±0.5μg. The cluster analysis were identified at the same cluster using liver resection sample and the super-thin needle aspiration sample.4. Final selection of genesAccording to our results above, gene expression profile of samples from hepatitis C and B were different, we consider that hepatitis C and B would be analyzed in different methods. At first, we analyzed the relationship of liver mean fibrotic change intensitiy (MFI) and gene expression profile.5. We analyzed a microarray analysis using 74 samples using super-thin-needle aspiration, then finally identified 39 genes that would be related liver damage of hepatitis C. Among them, MHC-class-II molecules were clearly reflected the inflammation status. Furthermore, stem cell activating genes, such as' PIK3C2B、 ARHGDIB, were included. These gene might be molecular-prognostic-factor of hepatitis patients.