罗哌卡因对骨肉瘤细胞增殖和凋亡的影响及作用机制  

Ropivacaine affects proliferation and apoptosis of osteosarcoma cells by down⁃regulating miR⁃199b⁃5p expression

作　　者：王铭[1] 司小萌[1] 陈欢[1] 郭培霞 张朔 梁玉柱[1] WANG Ming;SI Xiaomeng;CHEN Huan;GUO Peixia;ZHANG Shuo;LIANG Yuzhu(No.1 Operation Department,Nanyang City Center Hospital,Nanyang,Henan 473006,China)

机构地区：[1]南阳市中心医院手术一部,河南南阳473006

出　　处：《安徽医药》2020年第12期2347-2351,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨罗哌卡因对骨肉瘤细胞增殖和凋亡的影响及作用机制。方法体外培养骨肉瘤细胞MG63,分为对照组、低剂量罗哌卡因组、中剂量罗哌卡因组、高剂量罗哌卡因组、anti⁃miR⁃NC组、anti⁃miR⁃199b⁃5p组、中剂量罗哌卡因+miR⁃199b⁃5p组和中剂量罗哌卡因+miR⁃NC组。四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞增殖,流式细胞仪检测细胞凋亡,蛋白质印迹法(Western Blot)检测细胞中细胞周期蛋白D1(CyclinD1)、P21、B细胞淋巴瘤/白血病⁃2(Bcl⁃2)、Bcl⁃2相关X蛋白(Bax)表达水平,实时荧光定量逆转录PCR(RT⁃qPCR)检测细胞中miR⁃199b⁃5p表达水平。结果与对照组比较,低剂量罗哌卡因组、中剂量罗哌卡因组、高剂量罗哌卡因组MG63细胞增殖能力[(0.88±0.08)、(0.49±0.05)、(0.34±0.03)比(0.92±0.09)]、Cyclin D1蛋白[(0.85±0.08)、(0.66±0.06)、(0.41±0.04)比(0.95±0.09)]表达降低(P<0.05),而P21[(0.18±0.02)、(0.37±0.04)、(0.71±0.07)比(0.14±0.01)]蛋白表达升高(P<0.05),中剂量罗哌卡因组Bcl⁃2蛋白[(0.20±0.02)比(0.76±0.07)]及miR⁃199b⁃5p[(0.13±0.01)比(0.82±0.08)]的表达降低(P<0.05),而MG63细胞凋亡率[(23.51±2.42)%比(7.66±0.75)%]和Bax蛋白[(0.89±0.09)比(0.28±0.03)]表达升高(P<0.05)。与anti⁃miR⁃NC组比较,anti⁃miR⁃199b⁃5p组MG63细胞增殖能力[(0.64±0.06)比(0.98±0.09)]及Cyclin D1蛋白[(0.40±0.04)比(0.95±0.09)]和Bcl⁃2蛋白[(0.36±0.03)比(0.76±0.07)]的表达降低(P<0.05),而MG63细胞凋亡率[(19.56±1.58)%比(7.66±0.75)%]及P21[(0.53±0.05)比(0.14±0.01)]和Bax[(0.71±0.07)比(0.28±0.03)]的蛋白表达升高(P<0.05)。与中剂量罗哌卡因+miR⁃NC组比较,中剂量罗哌卡因+miR⁃199b⁃5p组MG63细胞增殖能力[(0.58±0.06)比(0.36±0.03)]及Cyclin D1蛋白[(0.71±0.07)比(0.40±0.04)]和Bcl⁃2蛋白[(0.53±0.05)比(0.21±0.02)]的表达升高(P<0.05),而MG63细胞凋亡率[(12.68±1.23)%比(23.35±2.34)%]及P21[(0.37±0.03)比(0.72±0.Objective To investigate the effect of ropivacaine on the proliferation and apoptosis of osteosarcoma cell line MG63 and its mechanism.Methods Osteosarcoma MG63 cells were cultured in vitro and assigned into control group,low⁃dose ropiva⁃caine group,medium⁃dose ropivacaine group,high⁃dose ropivacaine group,anti⁃miR⁃NC group,anti⁃miR⁃199b⁃5p group,medium⁃dose ropivacaine+miR⁃199b⁃5p group and medium⁃dose ropivacaine+miR⁃NC group.MTT method was used to detect cell prolifera⁃tion;flow cytometry was used to detect cell apoptosis;Western Blot was used to detect the protein expression level of CyclinD1,P21,Bcl⁃2 and Bax;RT⁃qPCR was used to detect the expression level of miR⁃199b⁃5p in cells.Results Compared with the control group,the proliferation ability of MG63 cells in the low⁃dose ropivacaine group,medium⁃dose ropivacaine group,and high⁃dose ropi⁃vacaine group[(0.88±0.08),(0.49±0.05),(0.34±0.03)vs.(0.92±0.09)]and the protein expression of Cyclin D1[(0.85±0.08),(0.66±0.06),(0.41±0.04)vs.(0.95±0.09)]decreased(P<0.05),while the protein expression of P21[(0.18±0.02),(0.37±0.04),(0.71±0.07)vs.(0.14±0.01)]increased(P<0.05);the expression of Bcl⁃2 protein[(0.20±0.02)vs.(0.76±0.07)]and miR⁃199b⁃5p[(0.13±0.01)vs.(0.82±0.08)]in the medium⁃dose ropivacaine group decreased(P<0.05),while the apoptotic rate of MG63 cell[(23.51±2.42)%vs.(7.66±0.75)%]and the expression of Bax protein[(0.89±0.09)vs.(0.28±0.03)]increased(P<0.05).Compared with the anti⁃miR⁃NC group,MG63 cells proliferation ability[(0.64±0.06)vs.(0.98±0.09)],the protein expression of Cyclin D1[(0.40±0.04)vs.(0.95±0.09)]and Bcl⁃2[(0.36±0.03)vs.(0.76±0.07)]in the anti⁃miR⁃199b⁃5p group decreased(P<0.05),while the apoptotic rate of MG63 cells[(19.56±1.58)%vs.(7.66±0.75)%]and the protein expression of P21[(0.53±0.05)vs.(0.14±0.01)]and Bax[(0.71±0.07)vs.(0.28±0.03)]increased(P<0.05).Compared with the medium⁃dose ropiva⁃caine+miR⁃NC group,MG63 cells proliferation ability[

关 键 词：骨肉瘤 罗哌卡因 细胞周期蛋白D1 基因 bcl⁃1 miR⁃199b⁃5p 细胞增殖 凋亡 

分 类 号：R738.1[医药卫生—肿瘤]