联合nrf2沉默与自噬抑制增强蛋白酶体抑制剂诱导的胰腺癌细胞ros依赖的线粒体凋亡

《联合nrf2沉默与自噬抑制增强蛋白酶体抑制剂诱导的胰腺癌细胞ros依赖的线粒体凋亡》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。

1、华中科技大学硕士学位论文激后，可抑制硼替佐米介导的自噬；硼替佐米诱导人胰腺癌细胞内ROS增多，抑制ROS后人胰腺癌细胞PARP1等凋亡相关蛋白、LC3B自噬标记蛋白及BIP、CHOP等内质网应激相关蛋白表达减少；硼替佐米诱导人胰腺癌细胞内NRF2、HO-1表达增加；联合NRF2沉默及自噬抑制后，人胰腺癌细胞细胞内ROS明显增加，且线粒体凋亡相关蛋白表达明显增加。结论：蛋白酶体抑制剂硼替佐米可抑制人胰腺癌细胞增殖；蛋白酶体抑制剂硼替佐米可通过增加人胰腺癌细胞内ROS水平诱导其线粒体凋亡。同时，由细胞内ROS增加

2、诱导自噬产生及NRF2表达增加可对抗ROS依赖的线粒体凋亡。同时沉默NRF2及自噬可增强硼替佐米引起的线粒体凋亡。关键词：自噬NRF2蛋白酶体抑制剂线粒体凋亡胰腺癌2华中科技大学硕士学位论文CombinationNRF2SilenceandAutophagyInhibitionEnhanceROS-dependentMitochondrialApoptosisinPancreaticCancerCellsInducedbyProteasomeInhibitorAbstractObjectiveToinvest

3、igatetheeffectofproteasomeinhibitorreactiveoxygenspeciesandapoptosisofhumanpancreaticcancercells.PreliminaryresearchcombinationNRF2silenceandautophagyinhibitionpromoteROS-dependentmitochondrialapoptosis.MethodsStudytheeffectofBoronbiologicalcharacteristicso

4、fpancreaticcancercells.CCK-8assayandcloneformationassaydetectedcellviability;AnnexinV-FITC/PIdoublestainingflowcytometryandwesternblottingtodetectcellapoptosis;WesternblottingdetectedtheexpressionofER-stressrelatedproteinsandnrf2;DCFH-DAfluorescentprobewere

5、usedtodetectreactiveoxygenspecies(ROS)level;JC-1fluorescentprobedetectcellmitochondriamembranepotential;Calciumlevelweredetectedbyusingfluo-3AMfluorescentprobe.DetectmechanismofBorresistanceinpancreaticcancerchemotherapy.changesofsubcellularstructureswereob

6、servedbyusingtransmissionelectronmicroscopy;thefluorescentautophagymakerGFP-LC3andthepositioningrelationshipofGFP-LC3andLyso-TrackerRedlysosomalprobewereobservedunderaconfocalmicroscopy;autophagywasinhibitedbyusingautophagyinhibitororsilencingatg5,andNRF2si

7、lencingwasusingRNAitechnology,andthenreactiveoxygenspecies(ROS)levelandapoptosiswereassessed.ResultBorinhibitedtheproliferationofhumanpancreaticcancercelllinesinadose-andtime-dependentmanner;Borinhibitedthecloneformationandinducedapoptosis;Bordecreasedmitoc

8、hondrialmembranepotentialofhumanpancreaticcancercells,andinduced3华中科技大学硕士学位论文mitochondrialapoptosis;BorcanincreasethenumberofautophagosomesandtheexpressionofLC3B,aautophagymakerprotein;Borinducedthefor