發(fā)布時(shí)間：2019-02-19 20:52

【摘要】：[目的]探討YB-1在低氧誘導(dǎo)的宮頸癌EMT發(fā)生中的作用及可能機(jī)制,為中晚期宮頸癌的靶向治療提供實(shí)驗(yàn)依據(jù)。 [方法](1)采用鏈霉菌抗生物素蛋白-過氧化酶(S-P)免疫組織化學(xué)方法檢測78例中、晚期宮頸癌組織微陣列芯片標(biāo)本中YB-1和Vimentin的表達(dá),探討其表達(dá)情況與各臨床病理特征的關(guān)系以及兩者的相關(guān)性。(2)應(yīng)用氯化鈷(CoCl2)處理宮頸癌Hela細(xì)胞模擬細(xì)胞低氧微環(huán)境,采用MTT法檢測CoCI2對Hela細(xì)胞增殖活力的影響,并篩選出最適作用濃度及作用時(shí)間。(3)CoCl2150μmol/L處理Hela細(xì)胞,6h后應(yīng)用倒置顯微鏡觀察Hela細(xì)胞的形態(tài)學(xué)特征；應(yīng)用細(xì)胞免疫熒光法檢測Hela細(xì)胞中YB-1、Vimentin和MMP-2的蛋白表達(dá)及其相互關(guān)系。 [結(jié)果] (1)YB-1在宮頸癌組織中的陽性表達(dá)率為30.8%(24/78),YB-1蛋白在宮頸癌組織中的表達(dá)與患者年齡(P=0.477)、腫瘤組織學(xué)類型(P=0.962)、淋巴結(jié)轉(zhuǎn)移(P=0.224)等無明顯相關(guān),而與FIGO分期和分化程度有關(guān)(P0.05)。Ⅳ期宮頸癌組織中YB-1蛋白的表達(dá)顯著高于Ⅱ、Ⅲ期(P=0.003)。高分化宮頸癌組織中YB-1的表達(dá)率為22.2%(4/18)；中分化宮頸癌組織中YB-1的陽性表達(dá)率為33.3%(17/51)；低分化宮頸癌組織中YB-1的陽性表達(dá)率為37.5%(3/8),隨著分化程度的降低YB-1的表達(dá)逐漸增高(P=0.001)。Vimentin在宮頸癌組織的陽性表達(dá)率為38.5%(30/78),Vimentin蛋白的表達(dá)與患者年齡無關(guān)(P=0.616),與腫瘤組織學(xué)類型(P=0.046)、、淋巴結(jié)轉(zhuǎn)移(P=0.004)、FIGO分期(P=0.020)和分化程度(P=0.000)密切相關(guān),且隨著FIGO分期的變晚和淋巴結(jié)轉(zhuǎn)移的出現(xiàn)Vimentin蛋白的表達(dá)逐漸增高(P0.05)。宮頸癌組織中YB-1和Vimentin表達(dá)呈正相關(guān)(r=0.375,P0.01)。 (2)CoCl2可明顯抑制Hela細(xì)胞的活力,并呈濃度-時(shí)間依賴性,模擬低氧微環(huán)境的CoCl2最適濃度為150μmol/L,最適作用時(shí)間為6h(P0.05)。 (3)低氧使Hela細(xì)胞逐漸獲得了間質(zhì)細(xì)胞的形態(tài)特征,150μmol/L的CoCl2處理Hela細(xì)胞6小時(shí)后,倒置顯微鏡觀察發(fā)現(xiàn)Hela細(xì)胞大部分逐漸伸展、變形,細(xì)胞間空隙變寬,突起變得更為狹長。 (4)低氧6h后,Hela細(xì)胞中YB-1、Vimentin、MMP-2表達(dá)較常氧組顯著增高(P0.05),且YB-1、Vimentin、MMP-2之間相互呈正相關(guān)(P0.05)。 [結(jié)論]YB-1可能通過YB-1-MMP-2-Vimentin途徑,上調(diào)Vimentin表達(dá)從而在低氧誘導(dǎo)的宮頸癌EMT發(fā)生中起到關(guān)鍵作用。
[Abstract]:[objective] to investigate the role and possible mechanism of YB-1 in hypoxic induced cervical cancer EMT, and to provide experimental evidence for the targeted therapy of advanced cervical cancer. [methods] (1) the expression of YB-1 and Vimentin were detected by streptavidin biotin peroxidase (S-P) immunohistochemical method in 78 cases of advanced cervical cancer. To investigate the relationship between the expression of CoCI2 and the clinicopathological features and the correlation between them. (2) to treat the hypoxic microenvironment of cervical cancer Hela cells with cobalt chloride (CoCl2), and to detect the effect of CoCI2 on the proliferation of Hela cells by MTT assay. The optimal concentration and time of action were obtained. (3) CoCl2150 渭 mol/L was used to treat Hela cells. After 6 hours, the morphological characteristics of Hela cells were observed by inverted microscope. The expression and correlation of YB-1,Vimentin and MMP-2 in Hela cells were detected by immunofluorescence assay. [results] (1) the positive expression rate of YB-1 in cervical carcinoma was 30.8% (24 / 78). The expression of YB-1 protein in cervical carcinoma was associated with age (P0. 477) and tumor histological type (P0. 962). The expression of YB-1 protein in stage 鈪，

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