中文摘要：

目的探讨槲皮素对宫颈癌细胞中乙酰肝素酶（HPA）表达的影响。方法免疫荧光法检测宫颈癌细胞系HeLa、Caski细胞中HPA蛋白的表达。实验分为3组，槲皮素组：在HeLa、Caski细胞中加入含不同浓度（20、40、80μmol／L）槲皮素的RPMI 1640培养基；阳性对照组：小分子RNA（siRNA）转染HeLa、Caski细胞沉默其HPA的表达后，加入RPMI1640培养基；空白对照组：在HeLa、Caski细胞中仅加入RPMI1640培养基。3组HeLa、Caski细胞分别培养24、48、72h后，采用实时定量PCR技术和蛋白印迹法检测其HPAmRNA和蛋白的表达。结果（1）免疫荧光法检测显示，HeLa和Caski细胞中均有HPA蛋白的表达，均位于细胞核与细胞质中，呈红色荧光。（2）实时定量PCR技术检测显示，槲皮素组HeLa、Caski细胞中HPAmRNA的表达水平均随槲皮素浓度（分别为20、40、80μmol／L）的升高、处理时间（分别为24、48、72h）的延长而明显下降（P〈0．01），呈浓度和时间依赖性。与空白对照组相比，槲皮素组中40、80μmol／L槲皮素处理后HeLa、Caski细胞中HPAmRNA表达水平均明显下降（P〈0．05）；与阳性对照组相比，槲皮素组中80μmol／L槲皮素处理后HeLa细胞中HPAmRNA表达水平无明显变化（P〉0．05），而Caski细胞中HPAmRNA表达水平明显升高（P〈0．01）。（3）蛋白印迹法检测显示，槲皮素组HeLa、Caski细胞中HPA蛋白的相对表达水平均随槲皮素浓度（分别为20、40、80μmol／L）的升高、处理时间（分别为24、48、72h）的延长而明显下降（P〈0．01），呈浓度和时间依赖性。与空白对照组相比，槲皮素组中40、80μmol／L槲皮素处理后HeLa、Caski细胞中HPA蛋白表达水平明显下降（P〈0．05）；与阳性对照组相比，槲皮素组中80μmol／L槲皮素处理后HeLa、Caski细胞HPA蛋白表达水平无明显变化（P〉0．05）。结论槲皮素对宫颈癌细胞中HPAmRNA和蛋白的表?

英文摘要：

Objective To detect the expression of heparanase （HPA） in cervical cancer cells and investigate the impact of quercetin on the expression of HPA, and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells. Methods The experimental groups included cervical cancer cell lines （HeLa and Caski） exposed to different concentrations of quercetin （20, 40 and 80 μmol/L） in the culture medium. The control groups included a negative control group, which was cultured with RPMI 1640 only, and a positive control group, in which cervical cancer cells were transfected with HPA small interference RNA （siRNA） to silence HPA expression. The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24, 48 and 72 hours after treatment. Results （1） HPA was significantly expressed in both cervical cancer cell lines （HeLa and Caski） , and it exists both nucleus and cytoplasm. （2）The real-time PCR shows as follows： as the quereetin concentration increased （20,40 and 80 μmol/L） , the mRNA expression level of HPA decreased （P 〈0. 01 ）, in which the inhibition of HPA expression was concentration dependent. In addition, the inhibition of HPA expression was also time dependent. As time growth, the expression level of HPA mRNA （24,48 and 72 hours） in HeLa and Caski ceils decreased （ P 〈 0.01 ）. Compared with negative control group, the expression level of HPA mRNA decreased in different concentrations of quercetin （40 and 80 μmol/L） in both HeLa and Caski cells （ all P 〈 0. 05 ） ; Compared with positive control group, the expression level of HPA mRNA expressed no obvious difference in quercetin （80 μmol/L） group （P 〉0. 05） in HeLa cells, while it was opposite in Caski cells（P 〈0. 01 ）. （3）The result of western blot shown that, as the quercetin concentration increased（20,40 and 80μmol/L）and time growth （24,48 and 72 hours）, the expression level of HPA protein dec