橙皮素抑制膀胱肿瘤细胞T24增殖及其机制研究  

The mechanism of hesperetin on proliferation inhibited in human bladder cancer

作　　者：曹智修[1] 袁敬东[1] 章传华[1] 周高峰[1] 黄遂斌[1] 白涛 吕磊[1] 黄韬[1] Cao Zhixiu;Yuan Jingdong;Zhang Chuanhua;Zhou Gaofeng;Huang Suibin;Bai Tao;Lyu Lei;Huang Tao(Department of Urology,Wuhan NO.1 Hospital,Wuhan 430022,China)

机构地区：[1]武汉市第一医院泌尿外科,430022

出　　处：《中华实验外科杂志》2022年第12期2319-2322,共4页Chinese Journal of Experimental Surgery

摘　　要：目的:探讨橙皮素对膀胱肿瘤细胞T24细胞株生长繁殖的影响,探讨橙皮素的相关作用机制。方法:体外培养购自美国模式培养无集存库人膀胱肿瘤T24细胞株,将2.5、5.0、10.0、20.0、40.0、60.0μmol/L橙皮素分别作用于T24细胞株24、48、72 h并通过噻唑蓝(MTT)法观察细胞生长和增殖状况;通过流式细胞仪检测细胞生长G_(0)期与G_(1)比值百分比的变化;并通过Transwell实验观察橙皮素对T24细胞迁移和侵袭的影响;蛋白质印迹法(Western blot)观察磷酸化细胞外信号调节激酶(p-ERK)、细胞外信号调节激酶(ERK)、磷脂酰肌醇3激酶(PI3K)、磷酸化蛋白激酶B(p-Akt)、蛋白激酶B(Akt)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)蛋白表达及与内参甘油醛-3-磷酸脱氢酶(GAPDH)比值的变化。两组之间的比较使用独立样本t检验,多组间比较使用单因素方差分析。结果:橙皮素可明显抑制T24细胞株的增殖,且经橙皮素处理24 h组的半抑制浓度(IC_(50))[(20.79±1.82)μmol/L]与处理48 h组的IC_(50)[(21.34±1.64)μmol/L]相当,两者差异无统计学意义(t=0.562,P>0.05),而两组高于72 h组(10.28±1.16)μmol/L,有统计学意义(t=10.498、11.060,P<0.05);10、20μmol/L组作用48 h后细胞周期G_(0)/G_(1)百分比分别为(70.12±5.21)%、(78.46±7.43)%,高于对照组[(57.86±5.39)%],差异有统计学意义(t=12.260、20.600,P<0.05);10、20μmol/L组作用48 h后侵袭抑制率分别为(25.37±1.76)%、(41.31±1.86)%,迁移的抑制率分别为(21.23±1.09)%、(34.67±1.58)%,高于对照组侵袭抑制率[(0.21±0.02)%]和迁移抑制率[(0.27±0.03)%],差异有统计学意义(t=25.170、41.110;t=21.030、34.470,P<0.05);10、20μmol/L组作用48 h后p-ERK、PI3K、p-Akt、N-Cadherin蛋白表达降低,与内参的比值分别为(0.46±0.06、0.44±0.06;0.54±0.08、0.51±0.09;0.48±0.06、0.47±0.07;0.50±0.05、0.49±0.05)比对照组与内参比值(0.67±0.06、0.82±0.11、0.71±0.04、0.78±0.10)低,差异有统计学意�Objective To investigate the effect of hesperetin on proliferation in human bladder cancer cell line T24,and reveal its relevant mechanisms.Methods Human bladder cancer T24 cells purchased from American type culture collection were treated with hesperetin at different concentrations(2.5,5.0,10.0,20.0,40.0,60.0μmol/L).The cell proliferation rate was assessed by methyl thiazolyl tetrazolium(MTT)assay,the cell cycle changes was observed by flow cytometry,the cell invasion and migration was observed by Transwell technology,and the expression changes of epithelial-mesenchymal transition(EMT)related protein was assessed by Western blotting.Results Hesperetin significantly inhibited proliferation of T24 cell,the half maximal inhibitory concentration(IC_(50))in treated for 24 h and 48 h groups were(20.79±1.82),(21.34±1.64)μmol/L respectively,which were significant different from being treated for 72 h group[(10.28±1.16)μmol/L,t=10.498,11.060,P<0.05].Hesperetin induced G_(0)/G_(1)cell cycle arrest,the ratio of G_(0)/G_(1)being treated for 48 h in 10 and 20μmol/L groups were(70.12±5.21)%,(78.46±7.43)%respectively,which were significant from control group[(57.86±5.39)%,t=12.260,20.600,P<0.05].Hesperetin inhibited T24 cell invasion and migration,the invasion inhibited ratio were(25.37±1.76)%,(41.31±1.86)%in 10 and 20μmol/L groups respectively,the migration inhibited ratio were(21.23±1.09)%,(34.67±1.58)%in 10 and 20μmol/L groups respectively,which were significant from control group[(57.86±5.39)%,t=25.170,41.110,t=21.030,34.470,P<0.05].The expression of phosphorylated protein kinase B(p-Akt),phosphorylated extracellular signal-regulated kinase(p-ERK),phosphatidylinositol 3 kinase(PI3K)and N-cadherin were decreased while E-cadherin increased.The expression ratio of these protein compared with GAPDH were 0.46±0.06,0.54±0.08,0.48±0.06,0.50±0.05,0.73±0.14 in 10μmol/L group respectively and were 0.44±0.06,0.51±0.09,0.47±0.07,0.49±0.05,0.74±0.20 in 20μmol/L group respectively.They were significant diffe

关 键 词：膀胱肿瘤 橙皮素 机制 

分 类 号：R737.14[医药卫生—肿瘤]