脉冲电场对胰腺癌细胞活性氧生成的影响  

Effect of pulsed electric field on reactive oxygen species generation in pancreatic cancer cells

作　　者：张至轩 李卓群 韩轩 张楚 杨海若 仵正[2] 吕毅[2] 张娜娜 Zhang Zhixuan;Li Zhuoqun;Han Xuan;Zhang Chu;Yang Hairuo;Wu Zheng;Lyu Yi;Zhang Nana(Institute of Regenerative and Reconstructive Medicine,Med-X Institute,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710049,China;Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院Med-X研究院再生与重建医学研究所,西安710049 [2]西安交通大学第一附属医院肝胆外科,西安710061

出　　处：《中华肝胆外科杂志》2023年第10期753-759,共7页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金青年项目(82202297);陕西省重点研发计划(2021LL-JB-06);西安交通大学第一附属医院科研发展基金(2021QN-23)。

摘　　要：目的探究不同强度的脉冲电场(PEF)作用下,胰腺癌细胞内活性氧(ROS)水平的变化。方法将鼠源胰腺癌细胞Panc02经电场强度分别为0、250、500、750、1 000 V/cm的PEF处理后,通过流式细胞技术和免疫荧光法,在体外探究细胞内ROS的生成规律,并探索不同场强下鼠源和人源胰腺癌细胞凋亡情况。选取20只6~8周龄的雄性C57BL/6 SPF级小鼠制备小鼠原位胰腺癌模型,分为5组,每组4只:250 V/cm PEF治疗组、500 V/cm PEF治疗组、750 V/cm PEF治疗组、1 000 V/cm PEF治疗组和假手术组。通过免疫荧光法检测各组小鼠残存肿瘤组织的ROS表达情况。结果在500 V/cm、750 V/cm、1 000 V/cm电场强度作用下,与作用后2 h相比,作用6 h、12 h、24 h、48 h后细胞内ROS表达阳性的细胞比例均降低,差异均具有统计学意义(均P<0.05)。与0 V/cm PEF处理相比,500 V/cm及750 V/cm PEF处理后的Panc02细胞的ROS均增加,差异具有统计学意义(均P<0.05);与相同作用时间下250 V/cm PEF处理相比,电场强度为500 V/cm和750 V/cm时Panc02细胞中ROS的表达水平均增加,差异具有统计学意义(均P<0.05)。Panc02细胞和MIA-PaCa-2细胞凋亡比例均随场强的增高而增加,并在场强为750 V/cm时,肿瘤细胞凋亡比例达到峰值。与假手术组相比,在500 V/cm PEF治疗组(16.65±6.01比2.38±1.21,t=-6.53)和750 V/cm PEF治疗组(16.54±4.41比2.38±1.21,t=-6.48)小鼠胰腺癌组织中ROS的表达量增加,差异均具有统计学意义(均P<0.001)。结论 PEF处理能够增加胰腺癌细胞及组织内ROS水平,当电场强度为500及750 V/cm时,ROS产生较多。Objective To investigate the changes of reactive oxygen species(ROS)in pancreatic cancer cells under the effect of pulsed electric fields(PEF).Methods Murine-derived pancreatic cancer cells Panc02 were treated with PEF at electric field strengths of 0,250,500,750,and 1000 V/cm,respectively.The intracellular ROS generation patterns under the different field strengths and at different times after the PEF were investigated in vitro by flow cytometry and immunofluorescence,meanwhile exploring the apoptosis of murine and human pancreatic cancer cells under different field strengths.Twenty 6-to 8-week-old male C57BL/6 SPF mice were prepared as orthotopic pancreatic cancer models and divided into five groups of four mice each:250 V/cm PEF group,500 V/cm PEF group,750 V/cm PEF group,1000 V/cm PEF group,and sham operation group.ROS expression in the residual tumor tissues of mice in each group was detected by immunofluorescence.Results Under the 500 V/cm,750 V/cm and 1000 V/cm electric field strength,the proportion of cells with intracellular ROS expression was decreased after 6 h,12 h,24 h and 48 h of the PEF compared with 2 h after the PEF,and the differences were statistically significant(all P<0.05).Compared with 0 V/cm PEF group,ROS expression increased in Panc02 cells treated with 500 V/cm and 750 V/cm PEF groups,and the differences were statistically significant(all P<0.05).Compared with 250 V/cm PEF group under the same time,ROS in Panc02 cells treated with 500 V/cm and 750 V/cm electric field strengths increased,and the differences were statistically significant(all P<0.05).The proportions of apoptosis of both Panc02 cells and MIA-PaCa-2 cells increased with rising field strength and peaked at the field strength of 750 V/cm.Compared with the sham-operated group,the expression of ROS was increased in pancreatic cancer tissues of mice in the 500 V/cm PEF-treated group(16.65±6.01 vs.2.38±1.21,t=-6.53)and 750 V/cm PEF-treated group(16.54±4.41 vs.2.38±1.21,t=-6.48),and the differences were statistically significan

关 键 词：脉冲电场 胰腺癌 电场强度 活性氧 

分 类 号：R735.8[医药卫生—肿瘤]