YB-1在低氧微环境诱导的宫颈癌EMT中的作用研究
发布时间：2019-02-19 20:52
【摘要】：[目的]探讨YB-1在低氧诱导的宫颈癌EMT发生中的作用及可能机制,为中晚期宫颈癌的靶向治疗提供实验依据。 [方法](1)采用链霉菌抗生物素蛋白-过氧化酶(S-P)免疫组织化学方法检测78例中、晚期宫颈癌组织微阵列芯片标本中YB-1和Vimentin的表达,探讨其表达情况与各临床病理特征的关系以及两者的相关性。(2)应用氯化钴(CoCl2)处理宫颈癌Hela细胞模拟细胞低氧微环境,采用MTT法检测CoCI2对Hela细胞增殖活力的影响,并筛选出最适作用浓度及作用时间。(3)CoCl2150μmol/L处理Hela细胞,6h后应用倒置显微镜观察Hela细胞的形态学特征；应用细胞免疫荧光法检测Hela细胞中YB-1、Vimentin和MMP-2的蛋白表达及其相互关系。 [结果] (1)YB-1在宫颈癌组织中的阳性表达率为30.8%(24/78),YB-1蛋白在宫颈癌组织中的表达与患者年龄(P=0.477)、肿瘤组织学类型(P=0.962)、淋巴结转移(P=0.224)等无明显相关,而与FIGO分期和分化程度有关(P0.05)。Ⅳ期宫颈癌组织中YB-1蛋白的表达显著高于Ⅱ、Ⅲ期(P=0.003)。高分化宫颈癌组织中YB-1的表达率为22.2%(4/18)；中分化宫颈癌组织中YB-1的阳性表达率为33.3%(17/51)；低分化宫颈癌组织中YB-1的阳性表达率为37.5%(3/8),随着分化程度的降低YB-1的表达逐渐增高(P=0.001)。Vimentin在宫颈癌组织的阳性表达率为38.5%(30/78),Vimentin蛋白的表达与患者年龄无关(P=0.616),与肿瘤组织学类型(P=0.046)、、淋巴结转移(P=0.004)、FIGO分期(P=0.020)和分化程度(P=0.000)密切相关,且随着FIGO分期的变晚和淋巴结转移的出现Vimentin蛋白的表达逐渐增高(P0.05)。宫颈癌组织中YB-1和Vimentin表达呈正相关(r=0.375,P0.01)。 (2)CoCl2可明显抑制Hela细胞的活力,并呈浓度-时间依赖性,模拟低氧微环境的CoCl2最适浓度为150μmol/L,最适作用时间为6h(P0.05)。 (3)低氧使Hela细胞逐渐获得了间质细胞的形态特征,150μmol/L的CoCl2处理Hela细胞6小时后,倒置显微镜观察发现Hela细胞大部分逐渐伸展、变形,细胞间空隙变宽,突起变得更为狭长。 (4)低氧6h后,Hela细胞中YB-1、Vimentin、MMP-2表达较常氧组显著增高(P0.05),且YB-1、Vimentin、MMP-2之间相互呈正相关(P0.05)。 [结论]YB-1可能通过YB-1-MMP-2-Vimentin途径,上调Vimentin表达从而在低氧诱导的宫颈癌EMT发生中起到关键作用。
[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，
本文编号：2426868
[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，
本文编号：2426868
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