L/O/G/OL/O/G/O

Yifeng Zhou Ph.Dzhouyifeng@suda.edu.cnzhouyifeng@suda.edu.cn

New Wine in Old BottlesNew Wine in Old Bottles

A New Approach of SNP Function ResearchA New Approach of SNP Function Research

Soochow Soochow UniversityUniversity

Why We Design This Study?Why We Design This Study?

What Did We Publish BeforeWhat Did We Publish Before?

What Did We not Publish What Did We not Publish Before?Before?

What Did We Perform Now?What Did We Perform Now?

What Did We Publish Now?What Did We Publish Now? 4

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An Unfinished Story: SNP in An Unfinished Story: SNP in TIMP2TIMP2 Promoters and Risk of Breast Cancer Promoters and Risk of Breast Cancer

My First PaperMy First Paper

Zhou Zhou et al.et al. Carcinogenesis.Carcinogenesis. 2004 25:399-404. 2004 25:399-404.

BackgroudBackgroud

TIMP-2: An Endogenous InhibitorTIMP-2: An Endogenous Inhibitor of MMP-2of MMP-2

TIMP-2 binds with Pro-MMP-2TIMP-2 binds with Pro-MMP-2

Result IResult I

Zhou Zhou et al.et al. Carcinogenesis.Carcinogenesis. 2004 25:399-404. 2004 25:399-404.

Result IIResult II

Zhou Zhou et al.et al. Carcinogenesis.Carcinogenesis. 2004 25:399-404. 2004 25:399-404.

Function Assays of SNPs in PromoterFunction Assays of SNPs in Promoter

SNPs in SNPs in PromoterPromoter

EMSAEMSA

ReportReportGeneGene

Real-TimeReal-TimePCR…PCR…

ChIPChIP

EMSAEMSA

Data UnpublishedData Unpublished

MechanismMechanism

Aishan HAN Aishan HAN et al.et al. ANALYTICAL SCIENCES 2006 ANALYTICAL SCIENCES 2006

MechanismMechanism

Huan Chen Huan Chen et al.et al. Biosensors and Bioelectronics 24 2009 1955–1961 Biosensors and Bioelectronics 24 2009 1955–1961

Material Material

GraphiteGraphite Glassy CarbonGlassy Carbon

IM6 systemIM6 system

Mechanism Mechanism

ResultsResults

(a)(a) Cyclic voltammograms (scan rate:100mVs-1) Cyclic voltammograms (scan rate:100mVs-1)

(b) Electrochemical impedance spectroscopy (Nyquist plots) response at different (b) Electrochemical impedance spectroscopy (Nyquist plots) response at different modified process in the electrode modified process in the electrode

MechanismMechanism

ResultsResults

Differential pulse voltammograms of [Fe(CN)6]3-/4-accumulated on theDifferential pulse voltammograms of [Fe(CN)6]3-/4-accumulated on the probe after its interaction with different concentration of the wtDNAprobe after its interaction with different concentration of the wtDNA

ResultsResults

ResultsResults

ResultsResults

The washing ratio responses as a function of NaCl concentrations in washing bufferThe washing ratio responses as a function of NaCl concentrations in washing buffer

ResultsResults

ResultsResults

TIMP-2TIMP-2 -418G/C -418G/C

Breast CancerBreast Cancer

ElectrochemistryElectrochemistry

Affinity of G is Affinity of G is significant higher than Csignificant higher than C

EMSAEMSA

No Difference No Difference Between two AlleleBetween two Allele

Case-ControlCase-Control

PP = 0.060 = 0.060

Report GeneReport Gene

G Allele G Allele significant significant higher than C Allelehigher than C Allele

ConclusionConclusion

• Present study provides further evidence Present study provides further evidence that the functional SNP in that the functional SNP in TIMP-2TIMP-2 promoter is genetic susceptibility factors for promoter is genetic susceptibility factors for breast cancer.breast cancer.

PublicationsPublications

Gao Y Gao Y et al.et al. Carcinogenesis. 2009 30:2064-9. Carcinogenesis. 2009 30:2064-9.

Acknowledgment Acknowledgment

• Yu XiaoYu Xiao

• Jiang LanJiang Lan

• Zheng JianZheng Jian

• Kang ShenhuiKang Shenhui

• Peng RuiPeng Rui

• Xu JinhaiXu Jinhai

L/O/G/OL/O/G/O

Thank You!zhouyifeng@suda.edu.cnzhouyifeng@suda.edu.cn