食管癌相關(guān)基因2（ECRG2）對腫瘤細(xì)胞增殖、遷移和轉(zhuǎn)移的抑制作用.pdf

1、中國防雨】醫(yī)科大學(xué)I尊士學(xué)位論文博士論文題目X761916食管癌相關(guān)基因2(ECRG2)對腫瘤細(xì)胞增殖、遷移和轉(zhuǎn)移的抑制作用博士生：黃革導(dǎo)師：陸士新專業(yè)：腫瘤學(xué)研究方向：腫瘤的細(xì)胞與分子生物學(xué)所在院所：腫瘤研究所入學(xué)時(shí)間：2002年9月2005年5月朱餐怍謄、導(dǎo)師船龠勿全文公布里墮型墮型奎堂堡主蘭垡簍奎重至Theinhibitionsofthetumorcellproliferation，migrationandmetastasisby

2、ECRG2invitroandinvivoABSTRACTEsophagealCancerRelatedGene2wasclonedandidentifiedinourlaboratoryPreviousdatashowthatECRG2hassimilareffectswithtumorsuppressorgene，buttheroleofECRG2intumordevelopmentandprogressandtheexactmec

3、hanismremainsunclearTogainfurthercomprehendthisissue，first，weclonedandsubclonedthefull1engthECRG2cDNAintothepGEX4TlvectortoproducedtherecombinantECRG2proteinjnEcoliandpurifiedECRG2GSTfusionproteinforfuturestudybyaffinity

4、chromatographyOilglutathionecoupledbeadsSecond，wegeneratedanti—ECRG2monoclonalantibodies(MAbs)byusingCterminalofECRG2peptideasimmunogen；theseantibodiesaresuitablefordetectingECRG2byELISAWesternblottingandlmmunofluorescen

5、cemicroscopyThird，wesuccessfullyconstrucledarecombinantplasmidpcDNA31一ECRG2andtransfecteditintoesophagealcancereelllineEC9706byusingLipofectamin2000TheresultsshowedthatthecolonyformationactivityofEC9706／pcDNA31一ECRG2wasl

6、8％whilethatofthecontrolcellEC9706／pcDNA31was55％(P005)111eactivityofanchorage—independentproliferationofEC9706／pcDNA3I—ECRG2waslowerthanthatofEC9706／pcDNA3IinsoftagarFourth，weextendedtheexaminationoftheroleofECRG2byusingr

7、ecombinantECRG2andsyntheticECRG2fragmentTIleEC9706cellsgrowthwassuppressedaftereelltreatedwiththesetweagentstoculturemedium；itconfirmedtheinhibitionofECRG2inproliferationoftumorcellsWhenEC9706cellswerepre—incubatedwithth

8、esetworeagentsandinoculatedSCintotheflankofmldemice，theoccHITenceofreplantedtumorswasdelayed，andgrowthoftumorswasjnhibitedTheseresultssuggestedthatECRG2inhibitedthetumoreellproliferationinbothinvitroandinvivoTheproteinho

9、mologousanalysisimplicatedthatECRG2containthebasicstructureofkazalproteaseinhibitorwhichmayplayakeyroleinthedegradationofECMcomponentsmediatingtumorjnvasionThestructuralresemblanceandd訊rerencebetweenECRG2andtheothermembe