Differential MicroRNA Expression Between Hepatitis B and
Hepatitis C Leading Disease Progression to Hepatocellular Carcinoma
HEPATOLOGY 2009;49:000-000 Yin-Hong Zhu
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Overview of Presentation Backgroud -why Protocol -how
Discussion-what Instruction-learn
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1 HBV,HCV,HCC HEPATOLOGY 2009;49:000-000. HCV HBV epidemiologic
findings 1998 Disease progression HCC promotion Genes involved in
inflammation Genes responsible for Anti-inflammation Pro-apoptotic
and DNA repair responses inammatory and anti-apoptotic phenotypes
Factors inducing those differences ? Signal pathwaysGenes
expression Signal pathways virologic featureshistologic findings
2001 2006Different oncogenic factors
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2 Regulation of gene expression microRNA 30% of human
genes
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3 Gene expression regulated by microRNAs.
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microRNA Development Proliferation (Stem cells) diseases such
as infection and cancer Differentiation Apoptosis 4 Function of
microRNA in animals and plants
6 microRNAs, HBV,HCV,HCC HBV or HCV Normal HBV or HCV,HCC HBV
between HCV,HCC mirRNA-122SPC3649 LNA-antimiRTM-122 Clinic trial
phase I on health person Blank
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7 Scientific supposition of this research HCV HBV microRNAs
Genes expression Signal pathways Disease progression HCC promotion
Dignosis,Therapy
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Overview of Presentation Backgroud-why Protocol -how Discussion
-what Instruction-learn
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1 Basic problems for miRNA researches What miRNAs are expressed
miRNA library selection Isolation,clone,sequence miRNA miRNA
profiles analysis Which gene is the target of the miRNAs Which
important miRNA influence Some certain bio-process When and where
the miRNAs are expressed Functional analysis Therapy Diagnosis
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2 Researches of microRNA and diseases miRNA Compare miRNA
profiles of diseases and normal samples miRNA Identify miRNAs that
affect cell division,apoptosis,angiogenesis,cell
proliferation,telomerrase activity miRNAs miRNAs Target genes
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3 Protocol for this research RTD-PCR Microarray Analysis.
Identification of microRNA Target genes Verication in HBV/ HCV
Infection Model Functional analysis miRNAs expressing profiles
analysis Verification of Regulation of genes by microRNA
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Normal RTD-PCR 3 Protocol for this research HCV HBV 12 patients
14 patients Control 9 normal liver tissue from patients with
metastatic liver cancer Hierarchical cluster analaysis Class
Prediction Binary tree calssification 13significantly differential
microRNAs 188 known miRNAs
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3 Protocol for this research Microarray Analysis. Global
analysis of mRNA expression in the liver tissues
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3 Protocol for this research Identification of microRNA Target
genes Predicted by MIRANDA Pro3.0 Step 1Step 2
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3 Protocol for this research Verification of Regulation of
genes by microRNA 2 Lost of funtcion transfect with Small
interfering RNAs specific to 13 candidate miRNAs 1 Gain of function
transfect Huh7.5 cells with Pre-miRNA miRNAs and Target genes mRNA
were detected by RTD-PCR
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3 Protocol for this research Verication in HBV/ HCV Infection
Model Huh 7.5 cellsJFH1-RNApHBV Virus proteins and DNA/RNA were
deteced miRNA expression
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Overview of Presentation Backgroud-why Discussion -What
Protocol -how Instruction -learn
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1 CH and HCC miRNAs in the same patient HEPATOLOGY
2009;49:000-000. HCV HBV No dependence Paired conditionDisease
Normal
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2 HBV and HCV infection- specific miRNAs HEPATOLOGY
2009;49:000-000. HCV HBV 6 miRNAs Cell death,DNA damage and
recombination, trancription signals 13 miRNAs Immune
response,antigen presentation, Cell cycle,proteasome,and lipid
metabolism Endogennous miRNA involved in defense mechanisms,mainly
RNA viruses Normal
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HEPATOLOGY 2009;49:000-000. 3 Tumor-associated miRNAs HCV HBV
has-miR-191 Worse survival in acute myeloid leukemia p53 tumor
supressor pathwayhas-miR-34 has-let-7a ras and c-myc genes
has-miR-133bSequamous cell carcinoma has-miR-142-5pHematopoietic
specific Normal
HEPATOLOGY 2009;49:000-000. 5 miRNAs in HBV/HCV disease
progression and HCC promotion
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Overview of Presentation Backgroud-why Instruction -learn
Protocol -how Discussion - What
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1 The same problem in differ- ent aspects Disease progression
HCC promotion HCV HBV microRNAs Genes expression Signal pathways
200120062009 Researches are still in need
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2 The same aspect with diffe- rent protocols Northern Blot
miRNarray qRT-PCR Luciferase reporter EMS A CHIP Western Blot 5RACE
MicroRNA Sensor reporter Bioinformatics prediction microarray
Cellular model Cellular invasion Cellular apoptosis Cellular
proliferation Cellular cycle qRT-PCR
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3 The same protocol for the similar researchs miRNA Compare
miRNA profiles of cancer and normal samples miRNA Identify miRNAs
that affect cell division,apoptosis,angiogenesis,cell
proliferation,telomerrase activity miRNAs miRNAs Target genes
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miRNA HEPATOLOGY 2009;49:000-000.
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qRT-PCR---miRNAs HEPATOLOGY 2009;49:000-000.
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miRNA siRNA
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MicroRNA
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miRNA HEPATOLOGY 2009;49:000-000.
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Q-rt-pcr HEPATOLOGY 2009;49:000-000.
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HEPATOLOGY 2009;49:000-000.
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microRNA HEPATOLOGY 2009;49:000-000.
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RTD-PCR 3 Protocol for this research HCV HBV 12 patients 14
patients Control 9 normal liver tissue from patients with
metastatic liver cancer Nomal