柚皮素通过调控糖原合成酶激酶3β/β-Catenin/Snaill信号通路对膀胱癌细胞增殖、凋亡的影响

Impacts of naringenin on proliferation and apoptosis of bladder cancer cells by regulating glycogen synthase kinase 3β/β-Catenin/Snail1 signaling pathway

目的石研究柚皮素通过调控糖原合成酶激酶3β(GSK-3β)/β-Catenin/Snaill信号通路对膀胱癌细胞增殖、凋亡的影响。方法采用CCK-8法检测0μg/ml、10μg/ml、20μg/ml、40μg/ml.60μg/ml、80μg/ml柚皮素处理的人膀胱癌细胞T24存活率,筛选出柚皮素的合适作用浓度。将体外培养的T24细胞随机分为对照组、柚皮素组、氯化锂组、柚皮素+氯化锂组,以柚皮素和氯化锂分组处理后,采用免疫印迹检测各组细胞GSK-3β/β-Catenin/Snaill信号通路蛋白表达;采用MTT法与细胞集落形成实验检测各组细胞增殖;采用流式细胞实验分别检测各组细胞凋亡;采用免疫荧光检测各组T24细胞增殖相关蛋白Ki67与凋亡相关蛋白Bcl-2、Bax表达。结果与对照组相比,柚皮素组细胞GSK-3β/p-GSK-3β、β-Catenin及Snaill蛋白表达、存活率、集落形成率、Ki67表达指数、Bcl-2/Bax、肿瘤重量、肿瘤体积降低(P<0.05),调亡率升高(P<0.05);氯化锂组细胞GSK-3β/p-GSK-3β、β-Catenin及Snaill蛋白表达、存活率、集落形成率、Ki67表达指数、Bcl-2/Bax、肿瘤重量、肿瘤体积升高(P<0.05);调亡率降低(P<0.05)。与柚皮素组相比,柚皮素+氯化锂组细胞GSK-3β/p-GSK-3β、β-Catenin及Snail1蛋白表达、存活率、集落形成率、Ki67表达指数、Bcl-2/Bax、肿瘤重量、肿瘤体积升高(P<0.05),调亡率降低(P<0.05)。结论仑柚皮素通过抑制GSK-3β/β-Catenin/Snaill通路进而抑制膀胱癌细胞增殖及其在裸鼠体内成瘤性,并促使其调亡。

Objective This paper aims to study the impacts of naringenin on the proliferation and apoptosis of bladder cancer cells by regulating the glycogen synthase kinase 3β(GSK-3β)/β-Catenin/Snaill signaling pathway.Methods CCK-8 method was applied to detect the survival rate of human bladder cancer T24 cells treated with naringenin at Oμg/ml,10μg/ml,20μg/ml,40μg/ml,60μg/ml,80μg/ml and screen the appropriate concentration of naringenin.T24 cells cultured in vitro were randomly grouped into control group and naringenin group,lithium chloride group group,naringenin+lithium chloride group;after treatment with naringenin and lithium chloride,the expression of GSK-3β/β-Catenin/Snaill signal pathway pro-tein was detected by Western blot;MTT assay and cell colony forming assay were applied to detect cell proliferation in each group;cell apoptosis was detected by flow cytometry;the expression of proliferation-related protein Ki67 and apoptosis related protein Bcl-2 and Bax of T24 cells in each group were detected by immunofluorescence.Results Compared with the control group,the expression of GSK-3β/p-GSK-3β,β-Catenin and Snaill proteins,survival rate,colony formation rate,Ki67 expression index,Bcl-2/Bax,tumor weight and tumor volume of naringenin group decreased(P<0.05),and the apop-tosis rate increased(P<0.05).The expression of GSK-3β/p-GSK-3β,β-Catenin and Snaill proteins,survival rate,colony formation rate,Ki67 expression index,Bcl-2/Bax,tumor weight and tumor volume increased in lithium chloride group(P<0.05);and the apoptosis rate decreased(P<0.05).Compared with naringenin group,the expression of GSK-3β/p-GSK-3β,β-Catenin and Snaill proteins,survival rate,colony formation rate,Ki67 expression index,Bcl-2/Bax,tumor weight and tumor volume increased in naringenin+lithium chloride group(P<0.05),and the apoptosis rate decreased(P<0.05).Conclusion Naringenin can inhibit the proliferation and tumorigenicity of bladder cancer cells in nude mice and pro-mote their apoptosis by inhibiting GSK-3β/β-Catenin/Snaill pathway.