大網(wǎng)膜乳斑巨噬細(xì)胞表型與不同分期胃癌關(guān)系的研究.pdf

1、大網(wǎng)膜乳斑巨噬細(xì)胞表型與不同分期胃癌關(guān)系的研究中文摘要目的：探討大網(wǎng)膜乳斑巨噬細(xì)胞(M巾)表型與不同分期胃癌的關(guān)系。方法：收集2011年3月至2011年12月青島市市立醫(yī)院不同分期胃癌患者大網(wǎng)膜乳斑標(biāo)本50例(其中T卜3期30例、T4期20例)以及正常人大網(wǎng)膜乳斑標(biāo)本10例。采用改良三步培養(yǎng)法及Ficoll密度梯度離心法純化培養(yǎng)巨噬細(xì)胞、倒置顯微鏡及瑞氏染色觀察細(xì)胞形態(tài)、CD68免疫熒光染色鑒定巨噬細(xì)胞、流式細(xì)胞術(shù)(FCM)檢測(cè)CD68

2、／CD206一M巾和CD206M2的表達(dá)情況、酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測(cè)不同時(shí)間點(diǎn)巨噬細(xì)胞IL一10、IL—12的分泌情況。對(duì)網(wǎng)膜乳斑巨噬細(xì)胞表型與胃癌病理學(xué)特征進(jìn)行相關(guān)性分析。結(jié)果：免疫熒光染色大網(wǎng)膜乳斑CD68巨噬細(xì)胞陽(yáng)性率為699％。流式細(xì)胞術(shù)檢測(cè)對(duì)照組、T13期組和T4期組大網(wǎng)膜乳斑CD68／CD206一M巾數(shù)量分別為(8295900546)X105個(gè)、(7453500546)X105個(gè)和(2652503002)105個(gè)

3、，三組差異具有顯著統(tǒng)計(jì)學(xué)意義(X2=49186，PO01)；對(duì)照組及T13期組CD68／CD206一M巾數(shù)量明顯高于T4期組為主(F=3418713、6305821，PO01)。大網(wǎng)膜乳斑CD206M2數(shù)量分別為(0169500242)X105個(gè)、(1006801213)X105個(gè)和(5914402958)105個(gè)，三組差異具有顯著統(tǒng)計(jì)學(xué)意義(x2=49194，PO01)。T4期組CD206M2數(shù)量明顯高于對(duì)照組及T13期組(Z=439

4、9、一5942，PO01)。CD68／CD206一M由與CD206M2表達(dá)及胃癌T分期呈明顯負(fù)相關(guān)(r=O586、一0832，PO01)，而CD206M2的表達(dá)與胃癌T分期呈明顯正相關(guān)(r=0826，P001)。ELISA檢測(cè)對(duì)照組、T13期組大網(wǎng)膜乳斑巨噬細(xì)胞不同時(shí)間點(diǎn)IL一10的平均分泌量均低于T4期組(均PO05)，而IL12平均分泌量均高于T4期組(均P005)。IL一12的分泌和ILIO的分泌及胃癌T分期呈明顯負(fù)相關(guān)(r一01

5、94、一0476，PO01)，而ILIO的分泌與胃癌T分期呈明顯正相關(guān)(r=0552，PO01)。在胃癌患者大網(wǎng)膜乳斑中，CD68／CD206一M中、CD206M2的表達(dá)與腫瘤是否侵犯漿Thestudyoftherelationbetweenthephenotypesofmacrophagesinmilkyspotsinhumangreateromentnmandgastriccarcinomaswithdifferentclinica

6、lstagesAbstractObjective：ToexploretherelationbetweenthephenotypesofmacrophagesinmilkyspotsinthegreateromentumandgastriccarcinomaswithdifferentstagesMethods：Specimensofhumangreatomentumofgastriccarcinomaswithdifferentstag

7、es(50casesof30instageT1—3and20instageT4)andnormalpeople(10cases)werecollectedinQingdaomunicipalhospitalfromMarch2011toDecember2011UsingamodifiedthreestepincubationmethodandFicolldensitygradientcentrifugationtoisolateandp

8、urifymacrophagesThecellswereidentifiedbymorphologicalobservationandimmunofluorescencestainingFlowCytometry(FCM)wasusedtodeterminetheexpressionofCD68／CD206‘a(chǎn)ndCD206macrophagesEnzymelinkedimmunosorbentassay(ELISA)Wasusedto

9、determinethesecretionofIL10andIL12indifferenttimepointsAndtherelmionbetweenthephenotypesofmacrophagesinmilkyspotsinthegreateromentumandthepathologicalfeaturesofgastriccarcinomaswasanalyzedResults：Theobtainedcellswereandw

10、ereconfirmedtobeCD68macrophageswithapositiverateof699％byimmunofluorescencestainingByFCMthenumberofCD68／CD206。MQinmilkyspotsingreateromentuminnormalcontrols，T1—3groupandT4groupwere(82959a：00546)105(74535士00546)105(59144士0

11、2958)105，respectivelyThenumberofCD68／CD206‘MQinnormalcontrolsandT13groupwaslargerthanthatininT4group(F=3418713、6305821，P001)Thereweresignificantdifferenceamongthetheregroups(f=49186，POo11ThenumberofCD206M2innormalcontrol