Using Pkc Regulated Mirnas as Core to Construct Hcc Suppression Network and to Search for Drugs to Restore This Network$S Function in Mirna Knockout Mice Induced Hcc Model

Stepwise progression of human hepatocellular carcinoma (HCC) has been well documented. To identify critical genes expression’s alterations accumulated during tumor progression is essential to understand the molecular mechanism of carcinogenesis and to design effective therapeutic strategy to cure this first cancer killer in Taiwan. Recent studies have shown a crucial role of miR101 in the developments of human prostate cancer and heptocellular carcinoma (HCC). Since regulatory role miRNA on gene expression is on both mRNA and protein level, how to integrate expression data generated from miRNA array, transcriptome and proteomics analysis to construct a realistic control network to lead developing novel therapeutic strategy become a new challenge. My laboratory recently has discovered that activation of protein kinase C in human hepatoma HepG2 cells induced growth arrest and up-regulated several miRNA expressions including miR101. Seven of TPA induced miRNA (101, 125b, 29c, 139-5p, 195, 150 and 497) were also found to be down regulated in human hepatoma tissues. We hypothesize that these coordinately regulated miRNA may act through altering both mRNA and protein level to form a tumor suppressor network. If we can construct putative miRNA mediated tumor suppressor network and to show collapsed network in HCC, we will be able to search for chemicals to restore gene expression and control network to normal in HCC. The potential of these chemicals to treat HCC should be further explored in both cultured cells and miRNA knockout induced HCC animal model. Therefore, in this proposal we set 4 specific aims as follows: 1, to understand how PKC coordinately up-regulates miR101 and six other miRNAs in HepG2 cells? 2, to develop a novel algorithm to construct miRNA-mediated tumor suppressor networks, including predicted miRNA-targets and downstream of targets based on datasets generated from miRNA, transcriptome and proteomics analysis analysis in both TPA treated HepG2 and 20 pair of HCC tissues. 3, to map gene signatures of miRNA-mediated tumor suppressor networks to Connectivity Map, a drug-associated gene expression profiles database, to identify small molecules which may mimic miRNA’s regulatory role to restore network function to normal and validate its potential as anti-HCC drug in cultured HCC cells and in miRNA knockout induced HCC animal model. 4, to establish miR101 knockout and HBX overexpression mice and to combine with miR122, p53 and Pten knockout as an animal model for liver cancer development.

Project ID:PA10101-1145
External Project ID:NSC99-2311-B182-004-MY3