title: miR-203a is involved in HBx-induced inflammation by targeting Rap1a.
abstract: Hepatitis B virus (HBV) causes acute and chronic hepatitis, and is one of the major causes of cirrhosis and hepatocellular carcinoma. Accumulating evidence suggests that inflammation is the key factor for liver cirrhosis and hepatocellular carcinoma. MicroRNAs play important roles in many biological processes. Here, we aim to explore the function of microRNAs in the HBX-induced inflammation. First, microarray experiment showed that HBV+liver samples expressed higher level of miR-203a compared to HBV-liver samples. To verify these alterations, HBx-coding plasmid was transfected into HepG2 cells to overexpress HBx protein. The real-time PCR results suggested that over-expression of HBx could induce up-regulation of miR-203a. To define how up-regulation of miR-203a can induce liver cells inflammation, we over-expressed miR-203a in HepG2 cells. Annexin V staining and BrdU staining suggested that overexpression of miR-203a significantly increased the cell apoptosis and proliferation, meanwhile, over-expression of miR-203a could lead to a decrease in G0/G1 phase cells and an increase in G2/M phase cells. Some cytokines production including IL-6 and IL-8 were significantly increased, but TGFβ and IFNγ were decreased in miR-203a over-expressed HepG2 cells. Luciferase reporter assay experiments, protein mass-spectrum assay and real-time PCR all together demonstrated that Rap1a was the target gene of miR-203a. Further experiments showed that these alterations were modulated through PI3K/ERK/p38/NFκB pathways. These data suggested that HBV-infection could up-regulate the expression of miR-203a, thus down regulated the expression of Rap1a and affected the PI3K/ERK/p38/NFκB pathways, finally induced the hepatitis inflammation.
authors: Department of gastroenterology, The First affiliated Hospital of Soochow University, Suzhou 215006, China.
authors: AiRong
authors: A
authors: Wu
authors: Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, China.
authors: Huo
authors: H
authors: Chen
authors: Department of gastroenterology, The First affiliated Hospital of Soochow University, Suzhou 215006, China.
authors: ChunFang
authors: C
authors: Xu
authors: Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, China.
authors: Ji
authors: J
authors: Zhou
authors: Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, China.
authors: Si
authors: S
authors: Chen
authors: Department of gastroenterology, The First affiliated Hospital of Soochow University, Suzhou 215006, China.
authors: YuQi
authors: Y
authors: Shi
authors: Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, China.
authors: Jie
authors: J
authors: Xu
authors: Department of gastroenterology, The First affiliated Hospital of Soochow University, Suzhou 215006, China. Electronic address: j_pzhang@suda.edu.cn.
authors: JianHe
authors: J
authors: Gan
authors: Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215123, China. Electronic address: ganjianhe@aliyun.com.
authors: JinPing
authors: J
authors: Zhang
mesh_terms: D017209:Apoptosis / Q000235:genetics; D002453:Cell Cycle; D049109:Cell Proliferation; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000502:physiology; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000473:pathology; D008099:Liver / Q000378:metabolism / Q000473:pathology / Q000821:virology; D020935:MAP Kinase Signaling System / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015534:Trans-Activators / Q000378:metabolism*; D054334:Viral Regulatory and Accessory Proteins; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D016207:Cytokines; C546703:MIRN203 microRNA, human; D035683:MicroRNAs; C430803:RAP1A protein, human; D012333:RNA, Messenger; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; D020827:rap1 GTP-Binding Proteins
keywords: HBx; Inflammation; MiR-203a; Rap1a