罗哌卡因通过AKT/GSK-3β/β-catenin信号通路对膀胱癌细胞生长和化疗效果影响  

Effects of ropivacaine on the growth and chemotherapy of bladder cancer cells through AKT/GSK-3β/β-catenin signaling pathway

作　　者：陈君[1] 李林艳[2] 赵晓勇 王鹏[2] 路云若雪 孙银贵[2] CHEN Jun;LI Linyan;ZHAO Xiaoyong;WANG Peng;LU Yunruoxue;SUN Yingui(School of Anesthesiology,Weifang Medical University,Weifang,Shandong261053,China;Department of Anesthesiology,Affiliated Hospital of Weifang Medical University,Weifang,Shandong261031,China)

机构地区：[1]潍坊医学院麻醉学院,山东潍坊261053 [2]潍坊医学院附属医院麻醉科,山东潍坊261031

出　　处：《中华肿瘤防治杂志》2023年第20期1208-1222,共15页Chinese Journal of Cancer Prevention and Treatment

基　　金：2021年山东省医药卫生科技发展项目(202104110339)。

摘　　要：目的探讨罗哌卡因(RVC)对膀胱癌细胞的生长和化疗效果的影响及其潜在作用机制。方法在使用不同浓度的罗哌卡因(0、0.1、0.5、1.0和2.0 mmol/L)处理膀胱癌细胞J82和T24细胞系后,采用CCK-8、EDU染色和细胞克隆形成能力实验检测细胞增殖水平。借助细胞划痕实验和Transwell小室检测罗哌卡因对细胞迁移和侵袭的影响。采用TUNEL和蛋白质印迹法检测细胞凋亡水平以及凋亡相关蛋白的表达。采用CCK-8检测罗哌卡因对丝裂霉素C(MMC)或吡柔比星(THP)的化疗效果影响。采用蛋白质印迹实验检测细胞AKT/GSK-3β/β-catenin信号蛋白的表达。结果在细胞增殖研究中,与对照组相比,随着罗哌卡因处理浓度的提高,膀胱癌J82(F=19.512,P<0.001)和T24细胞(F=68.939,P<0.001)增殖水平被显著抑制。随着罗哌卡因浓度升高,J82和T24细胞侵袭水平和迁移率均降低,F值分别为45.290、51.910、172.200和84.820,均P<0.001;J82(F=506.200,P<0.001)和T24细胞(F=811.500,P<0.001)凋亡率均升高;J82细胞Bcl-2(F=65.080,P<0.001)表达水平降低,Bax(F=91.150,P<0.001)和cleaved caspase-3(F=148.500,P<0.001)表达水平升高;T24细胞Bcl-2(F=96.320,P<0.001)表达水平降低,Bax(F=150.800,P<0.001)和cleaved caspase-3(F=91.660,P<0.001)表达水平升高。应用MMC后,随着罗哌卡因浓度升高,J82(F=195.630,P<0.001)和T24(F=108.390,P<0.001)细胞增殖水平均降低;应用THP后呈现出相同的趋势,F值分别为289.068和156.865,均P<0.001。随着罗哌卡因浓度升高,J82细胞p-AKT(F=41.770,P<0.001)和p-GSK-3β(F=40.030,P<0.001)表达水平均降低;T24细胞p-AKT(F=31.080,P<0.001)和p-GSK-3β(F=42.720,P<0.001)表达水平降低。加入MMC或THP后,罗哌卡因可通过AKT/GSK-3β/β-catenin信号通路影响细胞凋亡、侵袭和迁移,均P<0.001。结论罗哌卡因通过AKT/GSK-3β/β-catenin信号抑制膀胱癌细胞的生长,增强化疗效果。Objective To investigate the effects of ropivacaine(RVC)on the growth and chemotherapy of bladder cancer cells and its potential mechanism.Methods After the bladder cancer cell lines J82and T24were treated with different concentrations of ropivacaine(0,0.1,0.5,1.0and 2.0mmol/L),the cell proliferation was detected by CCK-8,EDU staining and cell clone formation ability test.The effects of ropivacaine on cell migration and invasion were detected using cell scratch experiments and Transwell cells.TUNEL and western blotting were used to detect the level of cell apoptosis and the expression of apoptosis related proteins.CCK-8was used to detect the effect of ropivacaine on the chemotherapy efficacy of mitomycin C(MMC)or pirarubicin(THP).The expression of cell AKT/GSK-3β/β-catenin signaling protein was detected using protein blotting assay.Results In the cell proliferation study,compared with the control group,with the increase of ropivacaine treatment concentration,the proliferation levels of bladder cancer J82(F=19.512,P<0.001)and T24cells(F=68.939,P<0.001)were significantly inhibited.As the concentration of ropivacaine increased,the invasion level and migration rate of J82and T24cells decreased,with F-values of 45.290,51.910,172.200and 84.820,respectively,all P<0.001;The apoptosis rates of J82(F=506.200,P<0.001)and T24cells(F=811.500,P<0.001)increased;The expression level of Bcl-2(F=65.080,P<0.001)in J82cells decreased,while the expression levels of Bax(F=91.150,P<0.001)and cleared caspase-3(F=148.500,P<0.001)increased;The expression levels of Bcl-2(F=96.320,P<0.001)in T24cells decreased,while the expression levels of Bax(F=150.800,P<0.001)and cleared caspase-3(F=91.660,P<0.001)increased.After the application of MMC,as the concentration of ropivacaine increased,the proliferation levels of J82(F=195.630,P<0.001)and T24(F=108.390,P<0.001)cells decreased;After application of THP,the same trend was observed,with F-values of 289.068and 156.865,both P<0.001.As the concentration of ropivacaine increased,the expression leve

关 键 词：膀胱癌 罗哌卡因 AKT/GSK-3β/β-catenin信号通路 增殖 迁移 凋亡 化疗 

分 类 号：R737.14[医药卫生—肿瘤]