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[槲皮素对子宫颈癌细胞中乙酰肝素酶表达的影响]

[Impact of quercetin on the expression of heparanase in cervical cancer cells].

作者信息

Zhang Wen-ting, Zhang Wei, Zhong Ya-juan, Lü Qiong-ying, Cheng Jing

机构信息

Department of Gynecology, Renmin Hospital of Wuhan University, Wuhan 430060, China.

出版信息

Zhonghua Fu Chan Ke Za Zhi. 2013 Mar;48(3):198-203.

Abstract

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 quercetin 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 cells 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 80 µmol/L) and time growth (24, 48 and 72 hours), the expression level of HPA protein decreased (P < 0.01), and the inhibition of HPA protein expression was concentration and time dependent. Compared with negative control group, the expression level of HPA protein 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 protein expressed no obvious difference in quercetin (80 µmol/L) group (all P > 0.05) in both HeLa cells and Caski cells (all P > 0.05).

CONCLUSION

Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines, which inhibition is concentration and time dependent.

摘要

目的

检测宫颈癌细胞中乙酰肝素酶(HPA)的表达,探讨槲皮素对HPA表达的影响以及槲皮素抑制宫颈癌细胞生长的分子机制。

方法

实验组包括在培养基中暴露于不同浓度槲皮素(20、40和80 μmol/L)的宫颈癌细胞系(HeLa和Caski)。对照组包括仅用RPMI 1640培养的阴性对照组,以及用HPA小干扰RNA(siRNA)转染宫颈癌细胞以沉默HPA表达的阳性对照组。在处理后24、48和72小时,用荧光定量实时PCR和蛋白质印迹分析检测HPA的细胞表达水平。

结果

(1)HPA在两种宫颈癌细胞系(HeLa和Caski)中均有显著表达,且存在于细胞核和细胞质中。(2)实时PCR结果如下:随着槲皮素浓度增加(20、40和80 μmol/L),HPA的mRNA表达水平降低(P < 0.01),其中对HPA表达的抑制呈浓度依赖性。此外,对HPA表达的抑制也呈时间依赖性。随着时间延长,HeLa和Caski细胞中HPA mRNA(24、48和72小时)的表达水平降低(P < 0.01)。与阴性对照组相比,HeLa和Caski细胞中不同浓度槲皮素(40和80 μmol/L)组的HPA mRNA表达水平均降低(均P < 0.05);与阳性对照组相比,HeLa细胞中槲皮素(80 μmol/L)组的HPA mRNA表达水平无明显差异(P > 0.05),而Caski细胞中则相反(P < 0.01)。(3)蛋白质印迹结果显示,随着槲皮素浓度增加(20、40和80 μmol/L)以及时间延长(24、48和72小时),HPA蛋白表达水平降低(P < 0.01),对HPA蛋白表达的抑制呈浓度和时间依赖性。与阴性对照组相比,HeLa和Caski细胞中不同浓度槲皮素(40和80 μmol/L)组的HPA蛋白表达水平均降低(均P < 0.05);与阳性对照组相比,HeLa细胞和Caski细胞中槲皮素(80 μmol/L)组的HPA蛋白表达水平均无明显差异(均P > 0.05)。

结论

槲皮素可抑制宫颈癌细胞系中HPA的表达,这种抑制呈浓度和时间依赖性。

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