circRNA MYLK基因干扰对胰腺癌PANC-1细胞线粒体膜电位、氧化损伤和微管形成的影响  

Effects of circRNA MYLK Gene Interference on Mitochondrial Membrane Potential,Oxidative Damage and Formation of Microtubules in Pancreatic Cancer PANC-1 Cells

作　　者：李青聪 黄鑫[1] 李自康[1] 李燕[1] 罗威 LI Qingcong;HUANG Xin;LI Zikang;LI Yan;LUO Wei(Department of Oncology,Ya'an People's Hospital(Ya'an Hospital,West China Hospital,Sichuan University),Ya'anSichuan 625000,China)

机构地区：[1]雅安市人民医院(四川大学华西医院雅安医院)肿瘤科,四川雅安625000

出　　处：《联勤军事医学》2024年第1期11-16,共6页Military Medicine of Joint Logistics

基　　金：吴阶平医学基金会临床科研专项资助基金(320.6750.2021-22-12)。

摘　　要：目的探讨circRNA肌球蛋白轻链激酶(myosin light chain kinase,MYLK)基因干扰对胰腺癌PANC-1细胞线粒体膜电位、氧化损伤和微管形成的影响。方法将对数生长期PANC-1细胞分为空白对照组、shRNA-NC组和circMYLK-shRNA组。转染后,采用细胞克隆形成实验检测各组PANC-1细胞的生长。试剂盒检测各组细胞上清液中超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)水平。流式细胞仪分析细胞线粒体膜电位。显微镜下观察各组细胞微管结节数。Western blot检测各组细胞Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)/B细胞淋巴瘤2基因(B-cell lymphoma 2,Bcl-2)、原癌基因c-Myc、血管内皮生长因子(vascular endothelial growth factor,VEGF)和波形蛋白(Vimentin)表达。结果与shRNA-NC组相比,circMYLK-shRNA1组PANC-1细胞的克隆形成率、JC-1红色荧光所占百分比、微管结节数明显降低(P均<0.05);细胞上清中SOD水平明显降低,MDA水平明显升高(P均<0.05);细胞内Bax/Bcl-2蛋白表达明显升高(P<0.05),c-Myc、VEGF和Vimentin蛋白表达明显降低(P均<0.05)。结论circRNA MYLK基因沉默可以抑制PANC-1细胞增殖能力,降低PANC-1细胞的线粒体膜电位,诱导细胞氧化损伤,抑制微管的形成。Objective To explore the effects of circRNA myosin light chain kinase(MYLK)gene interference on mitochondrial membrane potential,oxidative damage and formation of microtubules in pancreatic cancer PANC-1 cells.Methods PANC-1 cells in logarithmic growth phase were divided into blank control group,shRNA-NC group and circ-MYLK-shRNA group.After transfection,the growth of PANC-1 cells in each group was detected by clone formation as-say.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in supernatant of each group were detected by reagent.Mitochondrial membrane potential in each group was detected by flow cytometry.The number of microtubule nodules in each group was observed under the microscope.The expressions of Bcl-2 associated X protein/B-cell lymphoma 2(Bax/Bcl-2),proto-oncogene c-Myc,vascular endothelial growth factor(VEGF)and Vimentin in each group was detected by Western blot.Results Compared with shRNA-NC group,clonal formation rate,percentage of JC-1 red fluo-rescence and number of microtubule nodules of PANC-1 cells in circMYLK-shRNA1 group significantly decreased(all P<0.05);SOD level in cell supernatant significantly decreased,MDA level significantly increased(all P<0.05);expres-sion of Bax/Bcl-2 protein in cells significantly increased(P<0.05),and the protein expressions of c-Myc,VEGF and Vi-mentin significantly decreased(all P<0.05).Conclusion Silencing circRNA MYLK gene can inhibit the proliferation of PANC-1 cells,reduce mitochondrial membrane potential of PANC-1 cells,induce oxidative damage and inhibit formation of microtubules.

关 键 词：circRNA肌球蛋白轻链激酶 胰腺癌 线粒体膜电位 氧化损伤 微管 

分 类 号：R735.9[医药卫生—肿瘤]