【摘要】 目的 探究槲皮素对宫颈癌Hela/DDP细胞顺铂耐药性的影响及其可能作用机制。方法 体外培养人宫颈癌Hela/DDP、Hela细胞，按如下分组开展实验：(1)Hela/DDP、Hela细胞设置不同浓度(0、5、10、20、40、80μg/mL)顺铂处理组；(2)Hela/DDP细胞设置对照组(不处理)、顺铂组(10μg/mL顺铂)、低浓度槲皮素组(10μg/mL顺铂+20μmol/L槲皮素)、中浓度槲皮素组(10μg/mL顺铂+40μmol/L槲皮素)和高浓度槲皮素组(10μg/mL顺铂+80μmol/L槲皮素);(3)Hela/DDP细胞设置NC组、NC-siRNA组和miRNA-18a-siRNA组，按组转染处理，均加入终浓度为10μg/mL的顺铂和80μmol/L的槲皮素。CCK-8法检测Hela/DDP、Hela细胞增殖情况；流式细胞仪检测Hela/DDP细胞凋亡情况；实时荧光定量聚合酶链式反应法检测Hela/DDP细胞中miRNA-18a表达情况；蛋白免疫印迹法检测Hela/DDP细胞中B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达情况。结果 使用5、10、20、40、80μg/mL浓度的顺铂处理后，Hela/DDP细胞增殖抑制率均低于Hela细胞(P<0.05);顺铂对Hela、Hela/DDP细胞的半数抑制浓度(IC50)分别处于5～10μg/mL、10～20μg/mL之间。与对照组相比，顺铂组Hela/DDP细胞增殖率、细胞中Bcl-2蛋白表达水平显著降低(P<0.05),凋亡率、细胞中miRNA-18a、Bax蛋白表达水平显著升高(P<0.05);与顺铂组相比，低、中、高浓度槲皮素组Hela/DDP细胞增殖率、细胞中Bcl-2蛋白表达水平降低(P<0.05),凋亡率、细胞中miRNA-18a、Bax蛋白表达水平升高(P<0.05),且呈剂量依赖性(P<0.05)。与NC-siRNA组相比，miRNA-18a-siRNA组Hela/DDP细胞中miRNA-18a表达水平及细胞凋亡率均显著降低(P<0.05),细胞光密度(OD)值显著升高(P<0.05)。结论 槲皮素可在顺铂处理宫颈癌Hela/DDP细胞基础上进一步抑制细胞增殖并诱导细胞凋亡，且可能是通过上调miRNA-18a表达逆转宫颈癌细胞的顺铂耐药性。更多还原

【Abstract】 Objective To investigate the effect of quercetin on cisplatin resistance on cervical cancer cells Hela/DDP and its possible mechanism. Methods Hela/DDP and Hela cells were cultured in vitro, and perform experiments in groups as follows. First, Hela/DDP and Hela cells were treated with cisplatin at various concentrations(0, 5, 10, 20, 40, 80 μg/mL). Second, Hela/DDP cells were divided into control group(no treatment), cisplatin group(10 μg/mL cisplatin), low-dose quercetin group(10 μg/mL cisplatin + 20 μmol/L quercetin), medium-dose quercetin group(10 μg/mL cisplatin + 40 μmol/L quercetin) and high-dose quercetin group(10 μg/mL cisplatin + 80 μmol/L quercetin). Third, Hela/DDP cells were divided into NC group(no transfection), NC siRNA group(transfected with NC siRNA) and miRNA-18a-siRNA group(transfected with miRNA-18a-siRNA), followed by treatment with cisplatin(10 μg/mL) and quercetin(80 μmol/L). The proliferation of Hela/DDP and Hela cells was detected by CCK-8. assay. Real-time fluorescence quantitative PCR(qRT-PCR) was used to detect the expression of miRNA-18a in Hela/DDP cells. The apoptosis of Hela/DDP cells was detected by flow cytometry. Western blot was used to detect the expressions of Bcl-2 and Bax in Hela/DDP cells. Results After cisplatin induction, the proliferation inhibition rate of Hela/DDP cells was significantly lower than that of Hela cells(P<0.05). The half inhibitory concentration(IC50) of cisplatin was 5-10 μg/mL in Hela cells and 10-20 μg/mL in Hela/DDP cells. Compared with those in the control group, the proliferation rate and protein expression of Bcl-2 in Hela/DDP cells of the cisplatin group were significantly lower(P<0.05), while the apoptosis rate and the expression levels of miRNA-18a and Bax were significantly higher(P<0.05). Quercetin treatment dose-dependently inhibited the proliferation rate and protein expression of Bcl-2 in Hela/DDP cells(P<0.05), but increased the apoptosis rate, and expression levels of miRNA-18a and Bax(P<0.05), and showing a dose-dependent(P<0.05). Compared with those in NC siRNA group, expression level of miRNA-18a and the apoptosis rate of Hela/DDP cells in miRNA-18a-siRNA group were significantly lower(P<0.05). Conclusion Quercetin can inhibit the proliferation and induce apoptosis of cisplatin-induced Hela/DDP cells, and reverse cisplatin resistance of cervical cancer cells by up-regulating miRNA-18a.更多还原