槲皮素和乙酰肝素酶与宫颈癌Caski细胞侵袭与迁移机制的研究  

Effect of Quercetin and Heparin Enzyme on Invasion and Metastasis of Cervical Cancer Caski Cells

作　　者：程静[1] 张蔚[1] 吕琼莹[1] 张文婷[1] 钟亚娟[1] 

机构地区：[1]武汉大学人民医院,湖北武汉430060

出　　处：《实用妇产科杂志》2014年第3期212-215,共4页Journal of Practical Obstetrics and Gynecology

基　　金：国家自然科学基金项目(编号:31170329)

摘　　要：目的:通过Transwell小室基质侵袭和划痕损伤方法研究宫颈癌Caski细胞系侵袭和迁移能力,观察其与槲皮素、乙酰肝素酶(HPA)的关系。方法:根据GenBank数据库提供的人HPA基因核苷酸序列,按照小发夹RNA(ShRNA)设计原则设计4对特异性的寡核苷酸序列及阴性对照,分别克隆到pYr—1.1质粒载体中。将筛选出的质粒转染到宫颈癌Caski细胞中,并将Caski细胞分为Caski细胞组、Caski+槲皮素组、HPA—ShRNA—Caski组(HPA蛋白基因沉默的宫颈癌Caski细胞)和HPA-ShRNA-Caski+槲皮素组。然后进行Transwell小室基质侵袭和划痕损伤实验,观察不同时间宫颈癌细胞的侵袭和迁移能力。结果:①在6、12、24小时时,Caski+槲皮素组和HPA-ShRNA-Caski组划痕宽度明显大于Caski细胞组(P<0.05;P<0.01);HPA—ShRNA—Cas—ki+槲皮素组划痕宽度均大于其他3组(P<0.01)。()Caski+槲皮素组和HPA—ShRNA—Caski组有细胞侵袭能力的细胞数较Caski细胞组下降(P<0.05),HPA—ShRNA—Caski+槲皮素组较其他3组均明显下降(P<0.01)。结论:槲皮素对宫颈癌Caski细胞的侵袭和迁移能力有明显抑制作用,而HPA与宫颈癌细胞的侵袭、迁移过程有关。槲皮素对宫颈癌的侵袭和迁移能力的抑制作用可能与HPA相关。Objective：To evaluate the effect of heparan enzyme （heparanase HPA） and quercetin on in- vasion and metastasis of cervical cancer Caski cells through the transwell chamber and substrate scratch damage methods. Methods：According to the HPA gene nucleotide sequence in GenBank database, we de- sign 4 shRNA oligonucleotide sequences targeting HPA, and cloned into the pYr-l. 1 plasmid vector. One shRNA sequence was selected and transfected into cervical cancer Caski cells. Four groups were assigned as follows： control group, quercetin treatment group, HPA-ShRNA （group and HPA-ShRNA ＋ quercetin group. The invasion and metastasis abilityof cervical cancer cells were observed through Trans well chamber substrate scratch damage experiment. Results：After treatment duration of 6, 12, 24 hours, the migration a- bility in quercetin treatment group and HPA-shRNA group decreased significantly in comparison with control group （P 〈 0.05 and P 〈 0.01 ）, but no difference between quercetin treatment group and HPA-shRNA group（ P 〉0.05）. The migration ability in HPA-shRNA ＋ quercetin group decreased significantly in compari- son with the other 3 groups（ P〈0.01）. The invasion ability in quercetin treatment group and HPA-shRNA group decreased significantly in comparison with control group （P〈0.05）. The invasion ability in HPA-shR- NA ＋ quercetin group decreased significantly in comparison with the other 3 groups（ P 〈0.01 ） Conclusions： Quercetin could obviously inhibit the ability of invasion and metastasis of cervical cancer Caski cells. HPA was associated with the process of invasion and metastasis of cervical cancer cells, which might be involved in effect of quercetin.

关 键 词：宫颈癌 槲皮素 乙酰肝素酶 侵袭 迁移 

分 类 号：R737.33[医药卫生—肿瘤]