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1、染料木黃酮對(duì)卵巢癌HO【關(guān)鍵詞】卵巢腫瘤InhibitoryeffectofgenistEinonhumanovariancancercelllineHO8910PMinnudemice 【Abstract】AIM:ToinvestigatetheeffectsandpossiblemechanismsofgenistEIn(GEN),akindofphytoestrogens,binedanovariancancercellsinnudemice.METHODS:OvariancancercelllineHO8910PMd
2、erivedfromhumanovarianserousadenocarcinomaice,izedequallyinto4groups:GENtreatmentgroup,DDPtreatmentgroup,DDPplusGENtreatmentgroup,andacontrolgroup.Fourassofthenudemiceorphologyoftumorcellsicroscope.TheexpressionsofPAandmicrovasculardensity(MVD)inedbyimmunohistochemi
3、stry.RESULTS:GENandDDPeffectivelyinhibitedthegroanovariancancercellstransplantedintonudemiceandtheinhibitoryeffectorphologicalchangesincondensationandmarginationofnuclearchromatin,apoptoticbodiesandcellnecrosissignificantlyincreasedinGENgroupandGENplusDDPgroup.CONCL
4、USION:BothDDPandGENcaninhibitthegroanovariancancercellsinnudemiceandexertsynergisticantitumoreffect.TheanticancermechanismofGENmayinvolvesuppressingtheproliferationofovariancancercellsandangiogenesisandblockingthebloodsupplyofthetumor. 【Keys;genistein;cisplatin;neo
5、vascularization,pathologic;proliferatingcellnuclearantigen 【摘要】目的:觀察植物雌激素染料木黃酮(GEN)與順鉑(DDP)聯(lián)合應(yīng)用治療人卵巢癌裸鼠移植瘤的效應(yīng)及其機(jī)制.方法:建立20只人卵巢癌裸鼠移植瘤模型,隨機(jī)分成4組,分別為對(duì)照組、GEN組、DDP組及GEN+DDP組.4L/L小牛血清的RPMI1640培養(yǎng)液中,置于37℃,50mL/LCO2飽和濕度培養(yǎng)箱內(nèi),大量擴(kuò)增細(xì)胞.將處于對(duì)數(shù)生長(zhǎng)期的細(xì)胞制成2×107/mL懸液,注入裸鼠后背部皮下.穿刺點(diǎn)距注射部位1.2
6、cm,每只裸鼠皮下接種1點(diǎn),每點(diǎn)0.2mL,含4×106個(gè)細(xì)胞. 1.2.2治療待移植瘤直徑長(zhǎng)成4mm左右時(shí)(約接種后7d),采用隨機(jī)數(shù)字法將20只裸鼠隨機(jī)分成4組,每組5只.GEN組灌胃給予GEN50mg/kg・d,共30d;DDP組腹腔注射DDP5mg/kg・d,實(shí)驗(yàn)開(kāi)始連續(xù)用1L/次. 1.2.3觀察指標(biāo)GEN治療30d后,脫頸處死各組裸鼠,稱體質(zhì)量,解剖裸鼠.取移植瘤稱質(zhì)量,計(jì)算各組平均瘤質(zhì)量及抑瘤率(IR),IR=(1-治療組平均瘤質(zhì)量/對(duì)照組平均瘤質(zhì)量)×100%.按金正均法計(jì)算q值
7、來(lái)評(píng)價(jià)GEN與DDP兩藥合用的效果,q=(EA+B)/[EA+(1-EA)×EB],EA+B為兩藥合用時(shí)的抑瘤率,EA和EB為各藥單獨(dú)應(yīng)用時(shí)的抑瘤率.q<0.85表示兩藥有拮抗作用,0.85≤q≤1.15表示兩藥有相加作用,q>1.15表示兩藥有協(xié)同作用.各組裸鼠心、肝、脾、肺、腎等重要臟器及移植瘤,用40g/L中性甲醛溶液固定,常規(guī)石蠟包埋、切片.HE染色,光鏡下觀察.SP免疫組化按試劑盒說(shuō)明進(jìn)行操作,Ⅷ因子相關(guān)抗原抗體或PA抗體的工作濃度均為1∶100.Ⅷ因子只在血管內(nèi)皮細(xì)胞表達(dá),任何一個(gè)胞質(zhì)呈棕染色的內(nèi)皮細(xì)胞或內(nèi)皮細(xì)胞簇
8、,與周圍其他組織有明顯界限,無(wú)論有無(wú)管腔,定義為一個(gè)血管.在低倍鏡(100倍)下選定移植瘤內(nèi)血管密度最高的5個(gè)區(qū)域,再在高倍鏡(200倍)下計(jì)數(shù)每個(gè)視野的血管條數(shù),以平均值為該移植瘤的微血管密度(microvesseldensity,MVD)[4].PA定位于細(xì)胞核上,陽(yáng)性染