5-脫氧雜氮胞苷誘導(dǎo)肝癌細(xì)胞系分泌exosomes及相關(guān)免疫分子的研究.pdf

1、內(nèi)蒙古醫(yī)學(xué)院碩士學(xué)位論文5脫氧雜氮胞苷誘導(dǎo)肝癌細(xì)胞系分泌exosomes及相關(guān)免疫分子的研究姓名：薩仁高娃申請學(xué)位級別：碩士專業(yè)：人體解剖與組織胚胎學(xué)指導(dǎo)教師：吳巖20090501AStudyof5—Aza—deoxycytidineInducingHepatomaCarcinomaCellstoSecreteExosomesandImmunoAssociatedMoleculesAbstractExosomebiogenesisinv

2、olvestheinwardbuddingofmultivesicularbodies(MVB)toformintraluminalvesiclesWhenfusedwiththeplasmamembrane，theMVBreleasesthevesiclesintotheextracellularenvironmentasexosomesTheimportantfeatureofexosomesistheirOverallprotei

3、ncompositionsExosomesfromdifferentcellularsourcesbearspecificproteinsoftheiroriginalcelltypeTumor—derivedexosomescontaintumordifferentiationantigenssuchasMAGE，NY—ESO一1moleculeandheatshockproteinAsaresult，theycanplayarole

4、inthephysiologyoftheexosome—secretingcellandcouldplayanintercellularcommunicationpurposebyallowingexchangeofproteinsandlipidsbetweenthesecretingcellandtargetcellsTheycanbeprocessed，tested，andusedinamuchsimplerwaythancell

5、Objective5一Aza—deoxycytidine(5一Aza—CdR)Wasusedinducehepatomacarci—nonlaceilstosecreteexosomesSOastoinvestigateexosomesbionomics，itsimmunologicalfunctionsandtheanticanceractivityofsupportingimmuno—associatedmolecules，priv

6、idingexosomesasanewandeffectivecellfreevaccinefortumorimmunotherapyMethodsExosomesderivedfromHepG2cellsandHep3Bcellstreatedwith5—aza—CdRwereisolatedandpurifiedbyultrafiltrafioncentrifugationandsucrosegradientultracentrif

7、ugation，andthentheirultrastructuresandimmuno—moleculewereobservedbyelectronmicroscopyTheexpressionsofHSP70，HLA—IandNY—ESO一1wereinvestigatedbyWesternblot；theconcentrationofproteinsinexosomesWasdetectedbyBCATheexpressionof

8、P53Wasobservedbyreversetranscription—polymerasechainreaction(RT—PCR)ResultsThenumbersofexosomesandtheconcentrationoftotalproteinsinexosomeswereincreasedafterbeinginducedby5—aza—CdRAftertreatmentwith5一Aza—CdR，thecontainin

9、gofHSP70，HLA—IandNY—ESO一1moleculewereincreasedinHepG2cellsandHe：p3Bcells，whichwereconfirmedbyimmuno—electronandWesternblot(P005)，themRNAexpressionofwidetypep53genewasincreasedinbothHepG2cellsandHep3BcellsConclusion5一aza—