槲皮素诱导下乳腺癌细胞对三苯氧胺耐药的增敏效应

Quercetin-induced strengthening efficiency to Tamoxifen in Tamoxifen-resistance breast cancer cells

目的探讨槲皮素对逆转乳腺癌细胞内分泌耐药的应用价值。方法选用雌激素受体阳性的人乳腺癌细胞株MCF-7,高浓度短时间冲击法建立三苯氧胺(tamoxifen,TAM)耐药细胞株(MCF-7Ca/TAM-R)。根据槲皮素给药浓度将实验组MCF-7Ca/TAM-R细胞分为实验1组(10μmol/L)、实验2组(25μmol/L)和实验3组(50μmol/L),各浓度组再分成2个亚组:槲皮素+OHT(4-hydroxytamoxifen,4-OH-TAM)同时干预组、槲皮素干预12 h后加入OHT序贯干预组;阴性对照组加入OHT;空白对照组不加任何药物。噻唑蓝法(MTT)检测各组细胞的增殖抑制率,流式细胞术检测各组细胞的凋亡。结果高浓度短时间冲击法成功建立人乳腺癌三苯氧胺耐药细胞株。各实验组细胞均有不同程度的增殖抑制和凋亡(P=0.000),该效应随槲皮素诱导剂量的递增而增强(P=0.000);低剂量(10μmol/L)用药对耐药细胞的增殖和凋亡无显著影响(P>0.05和P>0.002),槲皮素和三苯氧胺同时或序贯给药差异亦无统计学意义(P>0.05和P>0.002)。结论在槲皮素诱导下,三苯氧胺对耐药细胞的增殖抑制和凋亡效应随槲皮素诱导剂量的增加而增强,提示槲皮素可能具有逆转乳腺癌细胞对三苯氧胺的耐药性,该效应与槲皮素和三苯氧胺同时或序贯给药无关。

[Objective] To explore the value of Quercetin in reversal resistance to endocrine in breast cancer cells. [Methods] The impact- method of high concentration and short-term to MCF- 7, the human breast cancer cell line with estrogen receptor-position were used to establish Tamoxifen-resistant cell strain （MCF- 7Ca/TAM- R）. According to the concentration of Quercetin, experimental group MCF- 7Ca/TAM-R cells were divided into experimental group 1 （10 Ixmol/L）, experimental group 2 （25umol/L） and group 3 （50 umol/L）. Each concentration group was further divided into two subgroups： Quereetin ＋ OHT （4- hydroxy Ta- moxifen, 4- OH- TAM） simul-administrating group and sequential intervening group with Quercetin inducing 12 hours followed by the OHT. Negative control group was addded OHT and blank control group without any drugs. Proliferative inhibitory rate of each group cells were detected by MTT and apoptotic rate by Flow Cy- tometry. [Results] Tamoxifen-resistant cells line of breast cancer had been established successfully with the impact- method of high concentration and short-term. All of the experimental group cells appeared prolifera- tive inhibition and apoptosis in different degrees （P =0.000）. These effects were enhanced with gradual in- creasing of induced dose of Quercetin （P=0.000）. Proliferative inhibition and apoptosis in low- dose （10 umol/ L） of Quercetin had no significant difference （P 〉0.05 and P 〉0.002）. The differences between simultaneous and sequential administration of Quercetin and Tamoxifen was no significant yet （P 〉0.05 and P 〉0.002）.[Conclusion] Under Quercetin induced, efficiency of proliferative inhibition and apoptosis of Tamoxifen- re- sistance cell were enhanced with gradually increasing dose of Quereetin. This result supports the presumplion that Quercetin was provided with reversal effect in Tamoxifen- resistance of breast cancer cells. This reversal effect is independent without administrative mode of simultaneity or sequence of Quercetin and Tamoxifen.