小鼠肝癌淋巴道轉移分子機制的探討.pdf

1、小鼠肝癌淋巴道轉移分子機制的探討碩士研究生二指導教師:指導小組:教授副教授教授專業(yè)名稱:侯力凌茂英辛毅呂申病理學飛中瘤轉移是一個復雜而有黑過WfyaF4llt月中瘤絕大多數(shù)的癌以淋巴道轉移為主，但因淋巴道轉移模型的缺少，癌淋巴道轉移機偉。尚知之甚少了目的用目建的小鼠肝癌高、低淋巴道轉移模型，探討其轉移的分子機制扮法將高低轉移小鼠肝癌”胞系二aF(高轉移)和HCaP(低轉移)在細胞培養(yǎng)環(huán)境中，加入淋巴結、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細胞后，以流式細胞儀動態(tài)檢測同側引流淋巴結內(nèi)淋巴細胞的細胞周期以TUNEL方法檢測了轉移淋巴結內(nèi)巨噬細胞凋亡情況以免疫組化Sp法檢測了r和P細胞中凋亡相關基因FasL、增殖相關基因PCNA凋亡抑制基因Bc:2的表達情況。結果在單純1640培養(yǎng)基中F和P細胞僅分泌微弱的MMP9，且二者分泌量無明顯差別。向培養(yǎng)基加入淋巴結勻漿后i細胞分泌大量UP9及其活性型，MMP2及

3、其活性型:P細胞分泌IMP9及其活性型、分泌%IP2量均明顯低于F細胞，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.流式細胞儀檢測發(fā)現(xiàn)F組小鼠引流琳巴結悶淋巴細胞增殖峰出現(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