丁哌卡因对胰腺癌PANC-1细胞氧化应激和凋亡的影响

Effect of bupivacaine on oxidative stress and apoptosis of pancreatic cancer PANC-1 cells

目的探究丁哌卡因对胰腺癌PANC-1细胞氧化应激和凋亡的影响。方法以不同浓度(0.07、0.15、0.3、0.6、1.25、2.5、5、10、20、40、80 mmol/L)丁哌卡因处理胰腺癌PANC-1细胞,采用CCK8试剂盒检测其对细胞增殖能力的影响,筛选实验浓度。采用TUNEL染色观察细胞凋亡,生化试剂盒检测氧化应激指标,荧光探针检测活性氧(ROS)含量,流式细胞术检测线粒体膜电位,实时荧光定量PCR和蛋白印迹检测生长标记物和细胞凋亡标记物的表达。结果与对照组比较,2.5、5 mmol/L丁哌卡因组p21 mRNA及蛋白表达、细胞凋亡率、上清液丙二醛(MDA)含量、ROS含量、JC-1单体百分比、Bax/Bcl-2、cleaved cas3/cas3水平增加,Survivin、Ki67 mRNA及蛋白表达、上清液SOD、GSH含量、c-Myc蛋白表达减少(P<0.05)。结论高浓度丁哌卡因可诱导胰腺癌PANC-1细胞增殖抑制和凋亡,提高细胞内氧化应激水平。

Objective To explore the effect of bupivacaine on oxidative stress and apoptosis of pancreatic cancer PANC-1 cells.Methods PANC-1 cells were treated with different concentrations of bupivacaine(0.07,0.15,0.3,0.6,1.25,2.5,5,10,20,40 and 80 mmol/L).The cell proliferation ability was detected by CCK8 kit and the cell apoptosis was observed by TUNEL staining.The oxidative stress indexes were detected by biochemical kits.The levels of reactive oxygen species(ROS)were detected by fluorescent probe.The mitochondrial membrane potential(MMP)was detected by flow cytometry.The expressions of growth markers and apoptosis markers were detected by real-time quantitative PCR and western blot.Results Compared with the control group,the expressions of p21 mRNA and protein,cell apoptosis rate,the levels of supernatant MDA and ROS,the percentage of JC-1 monomer,and Bax/Bcl-2 and cleaved cas3/cas3 levels were increased,while the levels of survivin,expression of Ki67 mRNA and protein,levels of supernatant SOD and GSH,and expression of c-Myc protein were decreased in 2.5 mmol/L and 5 mmol/L bupivacaine groups(P<0.05).Conclusion High-concentration bupivacaine can induce the proliferation inhibition and apoptosis of pancreatic cancer PANC-1 cells,and improve oxidative stress level.