布比卡因通过调节AMPK对人膀胱癌T24细胞恶性增殖、氧化应激及凋亡的影响  

Effects of bupivacaine on malignant proliferation,oxidative stress and apoptosis of human bladder cancer T24 cells by regulating AMPK

作　　者：陈明明 司马靓杰[1] 刘巍巍 张宜林[1] CHEN Mingming;SIMA Liangjie;LIU Weiwei;ZHANG Yilin(Department of Anesthesiology,the First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471003,China)

机构地区：[1]河南科技大学第一附属医院麻醉科,河南洛阳471003

出　　处：《广东药科大学学报》2020年第4期519-525,共7页Journal of Guangdong Pharmaceutical University

基　　金：河南省医学科技攻关项目(2018020922)。

摘　　要：目的探讨布比卡因对人膀胱癌T24细胞增殖、氧化应激和凋亡的影响。方法选择不同剂量的布比卡因处理人膀胱癌T24细胞,24 h后CCK8检测细胞存活率。选择无显著毒性剂量分别为0、1.25、2.5、5 mmol/L的布比卡因进行后续实验。克隆形成实验检测细胞增殖;RT-PCR检测增殖标记物的mRNA水平;流式细胞术检测细胞凋亡;试剂盒检测细胞SOD活性和MDA、GSH的含量;蛋白印迹法检测凋亡相关蛋白和AMPKα1、Ki67和Caspase-3的表达。结果T24细胞的存活率以布比卡因浓度依赖的方式降低,布比卡因浓度超过10 mmol/L时,T24细胞的存活率显著降低(P<0.05)。与布比卡因0 mmol/L组比较,布比卡因2.5 mmol/L组和布比卡因5 mmol/L组T24细胞克隆形成率、Ki67和Survivin mRNA表达、SOD活性和GSH含量均显著降低(P<0.05);细胞凋亡率、MDA含量、AMPKα1蛋白表达水平、Cleaved Caspase-3/Caspase-3和Bax/Bcl-2的蛋白表达比值均显著升高(P<0.05)。5 mmol/L布比卡因组细胞引入抑制剂Compound C后,SOD活性和Ki67蛋白表达水平均显著降低(P<0.05),AMPKα1蛋白表达水平和Cleaved Caspase-3/Caspase-3比值均显著升高(P<0.05)。结论布比卡因可通过上调AMPK水平抑制人膀胱癌T24细胞恶性增殖并诱导细胞氧化应激和凋亡。Objective To investigate the effects of bupivacaine on human bladder cancer T24 cell proliferation,oxidative stress and apoptosis.Methods Different doses of bupivacaine were used to treat T24 cells,and the cell survival rate was detected by CCK8 after 24 h.The dose of bupivacaine at 0,1.25,2.5 and 5 mmol/L without significant toxicity was selected for subsequent experiments,and cell proliferation was detected by the clone formation experiment.mRNA level of proliferation marker was detected by RT-PCR.Cell apoptosis was detected by flow cytometry.The contents of SOD activity,MDA and GSH were detected by the kits.The expression of apoptotic protein and AMPKα1 were detected by western blot.Results Bupivacaine treatment decreased the survival rate of T24 cells in a concentration-dependent manner.When the concentration of bupivacaine was over 10 mmol/L,the survival rate of T24 cells was significantly decreased(P<0.05).Compared with 0 mmol/L bupivacaine group,bupivacaine at the dose of 2.5 mmol/L or 5 mmol/L obviously decreased T24 cell clone formation rate,Ki67 and Survivin mRNA expression,SOD activity and GSH content(P<0.05).Meantime,bupivacaine treatment increased the apoptosis rate,MDA content,AMPKα1 expression,cleaved Caspase-3/Caspase-3 and Bax/Bcl-2 ratios(P<0.05).In the 5 mmol/L bupivacaine combined with compound C group,the activity of SOD and the expression level of Ki67 protein were significantly decreased,but the expression level of AMPKα1 protein and the ratio of cleaved caspase-3/caspase-3 were significantly increased.Conclusion Bupivacaine can inhibit the malignant proliferation of human bladder cancer T24 cells and induce cell oxidative stress and apoptosis by upregulating AMPK levels.

关 键 词：布比卡因 AMPK 膀胱癌 增殖 氧化应激 凋亡 

分 类 号：R737.25[医药卫生—肿瘤]