罗哌卡因抑制膀胱癌J82细胞增殖、迁移及侵袭的分子机制研究  

Molecular mechanisms of ropivacaine in inhibiting the proliferation,migration and invasion of bladder cancer J82 cells

作　　者：杨欣宇 杨阳 王立娟 YANG Xinyu;YANG Yang;WANG Lijuan(Department of Anesthesiology,Second Hospital of Baoding,Baoding,Hebei 071000,China;Department of Anesthesiology,Beijing Youlian Eye,Ear,Nose and Throat Hospital,Beijing 100000,China;Department of Otolaryngology,Second Hospital of Baoding,Baoding,Hebei 071000,China)

机构地区：[1]河北省保定市第二医院麻醉科,河北保定071000 [2]北京优联眼耳鼻喉医院麻醉科,北京100000 [3]河北省保定市第二医院耳鼻咽喉科,河北保定071000

出　　处：《检验医学与临床》2024年第21期3195-3200,共6页Laboratory Medicine and Clinic

基　　金：河北省卫生健康委员会科研基金项目(20242031);河北省保定市科学技术研究与发展指导计划(17ZF011)。

摘　　要：目的探讨罗哌卡因对人膀胱癌J82细胞增殖、迁移和侵袭的影响及对c-Jun氨基末端激酶(JNK)信号通路的调控作用。方法体外培养人膀胱癌J82细胞,该研究分为预实验和正式实验两个部分。预实验分组:对照组(不进行干预),低、中、高浓度罗哌卡因组(200、400、800μmol/L罗哌卡因);正式实验分组:对照组、罗哌卡因组(有显著作用浓度的罗哌卡因)、阳性药物组(100 nmol/L紫杉醇)、抑制剂组(罗哌卡因+20μmol/L JNK信号通路抑制剂SP600125)和激活剂组(罗哌卡因+1μg/mL JNK信号通路激活剂茴香霉素),干预24 h。分别采用细胞计数试剂盒-8(CCK-8)法、5-乙炔基-2′脱氧尿嘧啶核苷(EdU)、划痕法、Transwell小室法、蛋白免疫印迹法检测细胞活力、增殖率、迁移率、侵袭细胞数、上皮间质转化(EMT)及JNK通路相关蛋白水平。结果预实验中,与对照组比较,高浓度罗哌卡因组细胞活力明显降低(P<0.05),以高浓度罗哌卡因(800μmol/L)进行正式实验。正式实验中,罗哌卡因组和阳性药物组细胞增殖率、迁移率、侵袭细胞数,以及N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、纤维粘连蛋白(FN)和磷酸化(p)-JNK表达水平较对照组降低(P<0.05),E-钙黏蛋白(E-cadherin)表达水平较对照组升高(P<0.05)。与罗哌卡因组相比,抑制剂组细胞增殖率、迁移率、侵袭细胞数,以及N-cadherin、Vimentin、FN和p-JNK表达水平降低(P<0.05),E-cadherin表达水平升高(P<0.05),而激活剂组上述指标趋势则正好相反(P<0.05)。结论罗哌卡因可能通过阻断JNK信号通路活化抑制人膀胱癌J82细胞增殖、迁移、侵袭和EMT进程。Objective To investigate the effects of ropivacaine on the proliferation,migration and invasion of human bladder cancer J82 cells,as well as its regulatory effects on the c-Jun N-terminal kinase(JNK)signaling pathway.Methods Human bladder cancer J82 cells were cultured in vitro and the experiment of this study was divided into two parts:the preliminary experiment and the formal experiment.In the preliminary experiment,the groups were as follows:control group(without intervention),and low,medium,high concentrations of ropivacaine groups(200,400,800μmol/L ropivacaine).In the formal experiment,the groups included:control group,ropivacaine group(ropivacaine at the concentration with significant effects),positive drug group(100 nmol/L paclitaxel),inhibitor group(ropivacaine+20μmol/L JNK signaling pathway inhibitor SP600125),and activator group(ropivacaine+1μg/mL JNK signaling pathway activator fennel mycotoxin),with interventions lasting 24 h.Cell viability,proliferation rate,migration rate,number of invasive cells,levels of epithelial-mesenchymal transition(EMT)proteins and JNK signaling pathway-related proteins were detected by cell counting Kit-8(CCK-8)assay,5-ethynyl-2′-deoxyuridine(EdU)assay,wound healing assay,Transwell chamber assay and Western blot.Results In the preliminary experiment,compared with the control group,the cell viability was significantly reduced in the high concentration of ropivacaine group(P<0.05).Therefore,the formal experiment was conducted with high concentration ropivacaine(800μmol/L).In the formal experiment,the cell proliferation rate,migration rate,invasion cell number and N-cadherin,Vimentin,fibronectin(FN),phosphorylated(p)-JNK expression levels in ropivacaine group and positive drug group were lower than those in the control group(P<0.05),while the expression level of E-cadherin protein was higher than that in the control group(P<0.05).Compared with the ropivacaine group,the inhibitor group showed decreased cell proliferation rates,migration rate,invasion cell counts,and lower

关 键 词：膀胱癌 罗哌卡因 人膀胱癌J82细胞 c-Jun氨基末端激酶信号通路 增殖 迁移 侵袭 上皮间质转化 

分 类 号：R737.14[医药卫生—肿瘤] R318.16[医药卫生—临床医学]