亞硒酸鈉誘導胰腺癌PANC-1細胞凋亡及其機制的探討.pdf

1、溫州醫(yī)學院碩士學位論文亞硒酸鈉誘導胰腺癌PANC1細胞凋亡及其機制的探討姓名：向俊峰申請學位級別：碩士專業(yè)：外科學指導教師：張啟瑜201211溫州1醫(yī)學院碩士學位論文Ex)erimentalStu&theE—ofSodiumSeleniteongxoerlmentaStudvontheEftbctofSodiumSeleniteonlAOoDtosisofHumanPancreaticCancerCellSAbS仃actObjectiv

2、e：Toobservedtheeffectsofsodiumselenite(Na2Se03，SSE)onhumanpancreaticcancerPANC1cellsthroughinhibitionrate，apoptosisrateandcellmorphology，tostudythemechanismsofsodiumseleniteinducedapoptosisofhumanpancreaticcancerPANClcel

3、lsbydetectingthereactiveoxygenspecies，mitochondrialmembranepotential，cytochromeCandcaspase3changes，Methods：ThecellmorphologychangeofPANC一1cellsaftertreated、航msodiumselenitewasobservedbymicroscopecamera；Proliferationinhib

4、itedratewasdetectedbYMTTassaysTheDNAchangeswasobservedbyDNAladderextractionandelectrophoresisTheAnnexinVFITC／PIdoublestainingmethodwasusedtodetectcellapoptosisTheintracellularreactiveoxygenspecies(ROS)wasdetectedbYDCFH—D

5、AwithFluorescencemicroscopyandFlowcytometryThemitochondrialmembranepotentialwasdetectedbyJC一1withFlowcytometryEnzymelinkedimmuno—sorbentassay(ELISA)wasusedtoquantitativedetectionoftheapoptosisrelatedproteinCaspase3Wester

6、nblotwasusedtodetectproteincytochromeCreleasesion，GenecytCdetectbyFluorescencequantitativePCRResults：SodiumselenitesignificantlyinhibitthegrowthofhumanpancreaticcancerPANC1cellsaSconcentrationtimedependentwaymicroscopewa

7、sobservedwiththeconcentrationofsodiumseleniteincreased，theprolongationofPANC一1cellsbecameround，shrunken，floatingandbroken，deathrateincreasedgmduallYDNAladderelectrophoresisshowedtheladderribbon，accompaniedbythetail，sugge

8、stingthatalsoaccompaniedbyextensivenecrosisTheMTTaSsaysshowthat謝ththeconcentrationofsodiumseleniteincreasedandactiontimeextendedtheproliferation—inhibitedrateofPANC—lcellsincreasedgraduallyTheFlowCytometryillustratedthea

9、poptosisrate剛C1cellalsoprovedthisThelevelofintracellularreactiveoxygenspecies(ROS)andcaspase3contentofthecellundertreatedwithsodiumseleniteincreasedasaconcentrationdependentway；WesternBlotandRTPCRshowstheintracellularcyt