槲皮素对子宫颈癌HeLa细胞的作用及其机制  

Effects and its mechanism of quercetin on cervical cancer HeLa cells

作　　者：黄丽琼[1] 张蔚[1] 杨洋[1] 陶璐[1] 

机构地区：[1]武汉大学人民医院妇产科,430060

出　　处：《中华妇产科杂志》2009年第6期436-439,共4页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的观察中药槲皮素对宫颈癌HeLa细胞的生长抑制及促凋亡作用，并对其机制进行初步探讨。方法不同浓度（分别为6．25、12．5、25、50μmol／L）槲皮素作用24h后，采用四甲基偶氮唑蓝（MTT）比色法检测其对HeLa细胞生长的抑制作用，采用膜联蛋白V-异硫氰酸荧光素（FITC）／碘化丙啶（PI）双染色流式细胞术检测槲皮素作用24h后HeLa细胞凋亡率的变化。建立宫颈癌裸鼠模型并检测腹腔注射不同浓度槲皮素（分别为25、50、100mg·kg^-1·d^-1）15d后荷瘤裸鼠的抑瘤率。采用激光共聚焦荧光显微镜结合Ca^2＋荧光探针——Fluo-3／AM和荧光探针——JC-1，分别检测槲皮素作用后HeLa细胞内游离Ca^2＋（[Ca^2＋]i）浓度和线粒体膜电位的改变，并通过MTT比色法检测HeLa细胞内半胱氨酸天冬氨酸蛋白酶3（caspase-3）活性的变化。结果6．25、12．5、25、50μmol／L的槲皮素作用24h后，HeLa细胞生长的抑制率分别为（13．4±2．2）％、（26．2±6．8）％、（39．8±11．4）％、（48．5±9．1）％，凋亡率分别为（9．0±1．4）％、（13．3±1．1）％、（22．5±2．3）％、（44．7±4．2）％，其生长抑制率和凋亡率均呈明显的浓度依赖关系（P〈0．01）。25、50、100mg·kg^-1·d^-1的槲皮素对裸鼠移植瘤的抑瘤率分别为13．4％、30．4％和45．8％。随着槲皮素浓度的增加，HeLa细胞内线粒体膜电位降低、[Ca^2＋]i浓度升高、caspase-3活性增强，且均呈明显的浓度依赖关系（P〈0．01）。结论槲皮素能明显抑制宫颈癌HeLa细胞的生长；槲皮素可能通过激活细胞内Ca^2＋信号传导通路．最终通过线粒体徐径．诱导宫颈痛细胞的凋亡。Objective To evaluate the effects of quercetin on HeLa cells proliferation and study the mechanism of apoptosis. Methods The proliferation of HeLa cells treated with quereetin at different concentrations （ 6. 25, 12. 5,25,50 μmol/L ） for 24 hours was assessed by methyl thiazolyl tetrazolium （MTT） method, and also observe the inhibitory effect of quercetin on the transplanted carcinoma in BALB/ C nude mice. Cell apoptosis rate was detected by flow cytometry with annexin Vfluorescein isothiocyannate （FITC）/ propidium iodide （PI） technology. The changes of intracellular calcium ion concentration （ [ Ca^2＋ ] i） and mitochondrial membrane potential in HeLa cells induced by quercetin were studied by laser confocal fluorescence microscope and specificity fluorescent probes of Fluo-3/AM and JC-1. Colorimetric method was used to measure the activity of easpase-3. Results After treated by different concentrations of quercetin （6. 25, 12. 5, 25, 50 μmol/L） , the proliferation was inhibited and the apoptosis was induced in HeLa cells with the inhibition rate of （ 13.4 ± 2. 2 ） %, （ 26. 2 ± 6. 8 ） %, （ 39. 8 ± 11.4 ） % and （ 48. 5 ± 9. 1 ） % respectively, and with the apoptosis rate of （9.0 ± 1.4） %, （ 13.3 ± 1.1 ） %, （ 22. 5 ± 2. 3 ） % and （44. 7 ± 4. 2） % respectively, and which effects were dose-dependent （ P 〈 0. 01 ）. Treated by different concentrations of quercetin（25,50,100 mg· kg^- 1 · d^-1） , the growth of transplanted tumor in nude mice was inhibited with the inhibition rate of 13.4% ,30. 4% ,45.8%. The apoptosis of HeLa cells was induced by quercetin, which markedly reduced mitochondrial membrance potential, effectively enhance [ Ca^2 ＋ ] i, and the easpase-3 was activated in a dose-dependent manner （P 〈 0.01 ）. Conclusion Quercetin can significantly inhibit the proliferation of HeLa cells, which may be induced apoptosis of cervical cancer cells via the Ca^2＋ -dependent mitochondrial apoptosis pathway.

关 键 词：宫颈肿瘤 HELA细胞 槲皮素 细胞凋亡 

分 类 号：R686[医药卫生—骨科学]