小鼠肝癌淋巴道轉(zhuǎn)移分子機制的探討.pdf

1、小鼠肝癌淋巴道轉(zhuǎn)移分子機制的探討碩士研究生二指導(dǎo)教師:指導(dǎo)小組:教授副教授教授專業(yè)名稱:侯力凌茂英辛毅呂申病理學(xué)飛中瘤轉(zhuǎn)移是一個復(fù)雜而有黑過WfyaF4llt月中瘤絕大多數(shù)的癌以淋巴道轉(zhuǎn)移為主，但因淋巴道轉(zhuǎn)移模型的缺少，癌淋巴道轉(zhuǎn)移機偉。尚知之甚少了目的用目建的小鼠肝癌高、低淋巴道轉(zhuǎn)移模型，探討其轉(zhuǎn)移的分子機制扮法將高低轉(zhuǎn)移小鼠肝癌”胞系二aF(高轉(zhuǎn)移)和HCaP(低轉(zhuǎn)移)在細(xì)胞培養(yǎng)環(huán)境中，加入淋巴結(jié)、肝臟或脾臟勻漿，離心收集培養(yǎng)上清，

2、經(jīng)明膠酶譜法檢測各組的MMPs酶譜，分析酶譜的差異:經(jīng)小鼠皮下接種F和P細(xì)胞后，以流式細(xì)胞儀動態(tài)檢測同側(cè)引流淋巴結(jié)內(nèi)淋巴細(xì)胞的細(xì)胞周期以TUNEL方法檢測了轉(zhuǎn)移淋巴結(jié)內(nèi)巨噬細(xì)胞凋亡情況以免疫組化Sp法檢測了r和P細(xì)胞中凋亡相關(guān)基因FasL、增殖相關(guān)基因PCNA凋亡抑制基因Bc:2的表達(dá)情況。結(jié)果在單純1640培養(yǎng)基中F和P細(xì)胞僅分泌微弱的MMP9，且二者分泌量無明顯差別。向培養(yǎng)基加入淋巴結(jié)勻漿后i細(xì)胞分泌大量UP9及其活性型，MMP2及

3、其活性型:P細(xì)胞分泌IMP9及其活性型、分泌%IP2量均明顯低于F細(xì)胞，但不分泌MMP2的活性型。在加入肝臟或脾臟勻漿后，F(xiàn)和fr胞均不分泌AMPS.流式細(xì)胞儀檢測發(fā)現(xiàn)F組小鼠引流琳巴結(jié)悶淋巴細(xì)胞增殖峰出現(xiàn)晚二F尸繃.n.拼續(xù)時間很短，迅速滑落至低位，P組則始終ABSTRACTnvestigationonthemolecularbasisoflymphaticmetastasis時hepatocarcinomaHouLiLingMaoY

4、ingDepartmentofPathologyDalianMedicalUniversityNeoplasmmetastasisisacomplexandanactiveprocess.Lymphaticmetastasisisthemostnumerousofthemetastatictypeofcarcinomasbutthemolecularbasisoflymphaticmetastasisisstillunclearbeca

5、useoftheabsenceoflymphaticmetastaticmodel.ObjectiveToinvestigatethemolecularbasisoflymphaticmetastasisusing2groupsofmousehepatocarcinomacells(HcaFandHcaP)withdifferentlymphaticmetastaticpotential.MethodsThedifferenceofMM

6、Psproductionsandactivitiesbetweenthesetwocelllinesthatwereculturedtothemediumincludinghomogenateoflymphnodeorliverorspleentissueweredetectedwithzymographicanalysis.Thecellcycleofiymphcytesinlymphnodeofmicefollowinginject

7、ionsofFcelIsandPcellsweredetectedbyflowcytometryassay.WealsoexaminedtheapoptosisofmicrophagesinlymphnodesbyTUNEL.FasLPCNAandBcl2expressionsweredetectedinHcaFandHcaPcellsimmunohistochemicallv.ResultsHcaFandHcaPcellsproduc