Huang Li-Qiong, Zhang Wei, Yang Yang, Tao Lu
Department of Obstetrics and Gynecology, Renmin Hospital, Wuhan University, Wuhan, China.
Zhonghua Fu Chan Ke Za Zhi. 2009 Jun;44(6):436-9.
To evaluate the effects of quercetin on HeLa cells proliferation and study the mechanism of apoptosis.
The proliferation of HeLa cells treated with quercetin at different concentrations (6.25, 12.5, 25, 50 micromol/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 V- fluorescein isothiocyannate (FITC)/propidium iodide (PI) technology. The changes of intracellular calcium ion concentration ([Ca2+]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 caspase-3.
After treated by different concentrations of quercetin (6.25, 12.5, 25, 50 micromol/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 [Ca2+]i, and the caspase-3 was activated in a dose-dependent manner (P < 0.01).
Quercetin can significantly inhibit the proliferation of HeLa cells, which may be induced apoptosis of cervical cancer cells via the Ca2+ -dependent mitochondrial apoptosis pathway.
评估槲皮素对HeLa细胞增殖的影响并研究其诱导凋亡的机制。
采用噻唑蓝(MTT)法检测不同浓度(6.25、12.5、25、50 μmol/L)槲皮素处理HeLa细胞24小时后的增殖情况,并观察槲皮素对BALB/C裸鼠移植瘤的抑制作用。采用膜联蛋白V-异硫氰酸荧光素(FITC)/碘化丙啶(PI)技术通过流式细胞术检测细胞凋亡率。利用激光共聚焦荧光显微镜及Fluo-3/AM和JC-1特异性荧光探针研究槲皮素诱导HeLa细胞内钙离子浓度([Ca2+]i)及线粒体膜电位的变化。采用比色法检测半胱天冬酶-3的活性。
不同浓度(6.25、12.5、25、50 μmol/L)槲皮素处理后,HeLa细胞增殖受到抑制,凋亡被诱导,抑制率分别为(13.4±2.2)%、(26.2±6.8)%、(39.8±11.4)%和(48.5±9.1)%,凋亡率分别为(9.0±1.4)%、(13.3±1.1)%、(22.5±2.3)%和(44.7±4.2)%,且呈剂量依赖性(P<0.01)。不同浓度(25、50、100 mg kg(-1) d(-1))槲皮素处理后,裸鼠移植瘤生长受到抑制,抑制率分别为13.4%、30.4%、45.8%。槲皮素诱导HeLa细胞凋亡,显著降低线粒体膜电位,有效升高[Ca2+]i,且半胱天冬酶-3呈剂量依赖性激活(P<0.01)。
槲皮素可显著抑制HeLa细胞增殖,其可能通过Ca2+依赖的线粒体凋亡途径诱导宫颈癌细胞凋亡。