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目的:研究植物化学药物槲皮素对宫颈癌细胞的生长抑制作用和凋亡诱导效应,并探讨槲皮素对抗宫颈癌的可能作用机制。方法:采用浓度为10、20、40、80和160μmol/L槲皮素处理HeLa细胞,四唑盐比色法(MTT法)检测不同浓度在作用24、48和72h对HeLa细胞增殖的影响;流式细胞仪分析槲皮素对HeLa细胞凋亡的影响;逆转录-多聚酶链反应(RT-PCR)和蛋白质印迹法检测细胞内的MK和Caspase-3基因mRNA和蛋白表达水平。结果:槲皮素能抑制HeLa细胞增殖,且在一定浓度范围内呈时间浓度依赖性,P<0.05;使用浓度为10μmol/L的槲皮素处理HeLa细胞48h凋亡率为(2.18±0.04)%,20μmol/L为(3.75±0.11)%,40μmol/L为(6.04±0.07)%,80μmol/L为(9.65±0.04)%,160μmol/L为(21.41±1.81)%,与空白对组的(1.64±0.12)%比较差异有统计学意义,F=300.80,P<0.01;随槲皮素浓度的增高HeLa细胞中MK mRNA和蛋白表达依次下降,其中80和160μmol/L组下降较明显,而Caspase-3的mRNA和蛋白表达随药物浓度增高表达增强,P<0.01。MK与Caspase-3的蛋白表达呈负相关,r=-0.922,P<0.01;MK与Caspase-3的mRNA表达呈负相关,r=-0.955,P<0.01。结论:槲皮素可显著抑制HeLa细胞增殖并诱导其凋亡,其机制可能通过下调MK、上调Caspase-3的表达水平而发挥抗宫颈癌作用。
Objective: To study the inhibitory effect of phytochemical quercetin on cervical cancer cells and the induction of apoptosis, and to explore the possible mechanism of quercetin on cervical cancer. Methods: HeLa cells were treated with 10, 20, 40, 80 and 160μmol / L quercetin. The effects of different concentrations of HeLa cells on the proliferation of HeLa cells were detected by MTT method. The effect of quercetin on the apoptosis of HeLa cells was analyzed by flow cytometry. The mRNA and protein expression of MK and Caspase-3 in HeLa cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Results: Quercetin inhibited the proliferation of HeLa cells in a concentration-dependent manner (P <0.05). The apoptosis rate of HeLa cells treated with quercetin at the concentration of 10 μmol / L for 48 h was (2.18 ± 0.04) %, 20μmol / L was 3.75 ± 0.11%, 40μmol / L was 6.04 ± 0.07%, 80μmol / L was 9.65 ± 0.04%, 160μmol / L was 21.41 ± 1.81% (1.64 ± 0.12)%, the difference was statistically significant (F = 300.80, P <0.01). The expression of MK mRNA and protein in HeLa cells decreased with the increase of quercetin concentration, of which 80 and 160 μmol / L groups decreased more obviously , While the expression of Caspase-3 mRNA and protein increased with the increase of drug concentration, P <0.01. MK and Caspase-3 protein expression was negatively correlated, r = -0.922, P <0.01; MK and Caspase-3 mRNA expression was negatively correlated, r = -0.955, P <0.01. Conclusion: Quercetin can significantly inhibit the proliferation and induce apoptosis of HeLa cells. The mechanism may exert anti-cervical cancer effect by down-regulating MK and up-regulating the expression of Caspase-3.