羽扇豆醇联合槲皮素对膀胱癌5637细胞的抑制作用

Inhibitory effect of lupeol and quercetin on bladder cancer 5637 cells

目的研究羽扇豆醇(Lupeol)联合槲皮素(Quercetin)对膀胱癌5637细胞增殖抑制的作用,为膀胱癌的治疗提供实验依据。方法利用噻唑蓝(MTT)法分别检测不同浓度的羽扇豆醇(10、20、40、60、80、100、120μmol/L)及槲皮素(5、10、20、40、60、80μmol/L)对膀胱癌5637细胞的增殖抑制作用,并计算作用48 h后细胞的生长抑制率。选取抑制率为30%的药物浓度(即IC^(30))作为加药浓度分别单独及联合处理5637细胞48 h,MTT法检测各处理组细胞生长抑制率,并计算两种药物多个组合的Q值,以评价联合效果。利用克隆形成实验及划痕实验检测细胞克隆能力及迁移能力的影响。利用实时荧光定量反转录PCR(qRT-PCR)检测miR-145的表达水平。结果羽扇豆醇和槲皮素均能有效抑制膀胱癌细胞的增殖,呈浓度依赖性,槲皮素的量效变化更为明显。计算得出羽扇豆醇和槲皮素48 h的IC50分别为50.02μmol/L和44.42μmol/L;IC^(30)分别为35.40μmol/L(实验中使用35μmol/L)和29.86μmol/L(实验中使用30μmol/L),羽扇豆醇与槲皮素联合作用5637细胞抑制率达58.83%,两种药物为协同作用(Q值为1.167)。两种药物联合作用可明显抑制5637细胞的克隆形成及迁移能力,并使miR-145相对表达量增高。结论羽扇豆醇和槲皮素均能抑制膀胱癌5637细胞生长,并呈浓度依赖性,两种药物联合能协同抑制细胞的增殖,并明显抑制其克隆形成能力及迁移能力,其分子机制可能与miR-145的表达增高相关。

Objective To study the effects of Lupeol combined with quercetin on the proliferation inhibition of bladder cancer 5637 cells,and to provide experimental evidence for the treatment of bladder cancer.Methods The MTT method was used to detect different concentrations of lupeol(10,20,40,60,80,100,120μmol/L)and quercetin(5,10,20,40,60,80μmol/L)Inhibiting the proliferation of bladder cancer 5637 cells,and calculating the growth inhibition rate of the cells after 48 h.The drug concentration with the inhibition rate of 30%(IC^(30))was selected as the dosing concentration to treat 5637 cells individually and in combination for 48 h.The MTT method was used to detect the cell growth inhibition rate of each treatment group,and the Q value of multiple combinations of the two drugs was calculated and evaluated the joint effect.The effects of cell cloning ability and migration ability were detected by clone formation experiment and scratch experiment.The expression level of mir-145 was detected by real-time fluorescence quantitative reverse transcription PCR(qRT-PCR).Results Both lupeol and quercetin can effectively inhibit the proliferation of bladder cancer cells in a concentration-dependent manner,and the dose-effect change of quercetin was more obvious.The IC50 of lupeol and quercetin at 48 h was 50.02μmol/L and 44.42μmol/L respectively.The IC^(30) of lupeoland quercetin were 35.40μmol/L(35μmol/L in the experiment)and 29.86μmol/L(30μmol/L in the experiment)respectively.The inhibition rate of 5637 cells under the combined treatment of lupeol and quercetin was 58.83%,and the two drugs had a synergistic effect(Q value was 1.167).The combination of the two drugs could significantly inhibit the cloning and migration ability of 5637 cells,and increase the relative expression of miR-145.Conclusion s Both lupeol and quercetin could inhibit the growth of bladder cancer 5637 cells in a concentration-dependent manner.The combination of the two drugs could synergistically inhibit the cell proliferation,and significantly inhibit cell cloning and migration ability.The molecular mechanism might be related to the increased expression of miR-145.