線粒體DNA D-Loop區(qū)基因突變、ROS水平與肝癌關(guān)系的研究.pdf

1、東南大學(xué)碩士學(xué)位論文線粒體DNADLoop區(qū)基因突變、ROS水平與肝癌關(guān)系的研究姓名：黃學(xué)文申請學(xué)位級別：碩士專業(yè)：遺傳學(xué)指導(dǎo)教師：張麗珊20030818東南大學(xué)碩士學(xué)位論文RelationshipbetweenhepatocellularcarcinomaandmutationsintheD—LoopregionofmitochondrialDNAandtheROSlevelGeneticResearchCenterMedicalCo

2、flege，SoutheastUniversityPostgraduate：HUANGxuewenTutor：ZHANGli—shanobiectiveToinvestigatemutationsintheD—LoopregionofmitochondrialDNAinDfimaryhepatocellularcarcinoma，andmeasure】evelsofROSandMnSODinserumandtissuresofprima

3、ryhepatocellularcareinomaInordertogettherelationshipbetweenROSandroutationinDLoopweinvestigatefunetionsamongthemutationsinmitochundrialDNAROSandMnSODinthetumorMethodsTheDLoopregionof20hepatocellularscalcinonlatighterwith

4、theadjacentllOlnmltissueswereamplifiedbyPCRandsequencedMeanwhilethereactiveoxygenspecies(ROS)levelinserumweredetectedbyFentonreactionandGriessPrincipleROSintissueweremeasuredbyflowcytometryTSODandMnSODlevelinSelumandtiss

5、ueweredetectedbythemethodofxanthincoxidasercontrolgroupisthesenlmofnormalmanintheprocessofdetectingROSandMnSODinserumandcontrolgroupisthenormaltissuesadjacentthetumortissueintheprocessofdetectingROSandMnSODintissue)Resul

6、tsAmongthe20pairsofhepatocellulars，8pairsofmutationswerewiththemutationrate40％Therewere53mutationsdetectedinthegPairshepatocellulars，containing2insretions，11deletionsand40pointmutations75％pointmutationswereTCandCTtransit

7、ionThereare4microsatelliteinstabilitiesamongthemutationsMutationsintheadjacentnormaltissuesalwaysfo10wmutationsidentifiedintumortissuestherateofmutationsinthetumortissueishigherthantherateoftheadjacentnormaltissuesandalt

8、ofthemutationsidentifiedherewerehomoplasmicTherewerealargerdeletionhavebeenidentifiedTheROS1evelintheserunlandtissueofhepatocellularcarcinomaismuchhigherthanthecontrolgroup(p001)buttheTSODM11SODlevelintheseFumofhepatocel

9、lularcaicinomaismuchlowerthanthecontrolgroup(p001)Intheserumofpatient，theROSlevelisnegativecorrelationwiththeMnSODlevel(t=lO27，F(xiàn)一0924，pO01)11le■SOD，h佃SODleveIinthetissues’ofhepatocellularcarcinomaismuchhigherthanthecontr

10、olgmup(p001)Meanwhile。wefindtheROS1evelofhepatocellularcarcinomatissuewhichmtDNAD—LoopmutatedishigherthanthetheROSlevelofhepatocellularcarcinomatissuewhichmtDNAD—Loopdidn’tmutate，thel的Slevelintissuesofadjacenthepatocellu

11、larcarcinomawhichmtDNADLoopmutatedishigherthantheROSlevelintissuesofadjacenthepatocellularcareinomawhichIntDNADLoopdidn’tmutatedConclusions1TheDLoopregionofmitochondrialDNAisahighlypolymorphoricandmutableregionThemutatio