高压氧增强肺癌A549细胞对常见肺癌化疗药物敏感性研究  

Hyperbaric oxygen enhancing the sensitivity of lung cancer A549 cells to common lung cancer chemotherapy drugs

作　　者：刘维萍[1] 张丽芳[2] 陈云[1] 弓清梅[1] Liu Weiping;Zhang Lifang;Chen Yun;Gong Qingmei(Department of Respiratory and Critical Care Medicine,Shanxi Provincial People's Hospital,Taiyuan 030012,China;Department of Emergency,Shanxi Provincial People's Hospital,Taiyuan 030012,China)

机构地区：[1]山西省人民医院呼吸与危重症医学科,太原030012 [2]山西省人民医院急诊科,太原030012

出　　处：《中华航海医学与高气压医学杂志》2022年第4期478-482,共5页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：山西省卫生计生委科研课题(2017019)。

摘　　要：目的:探讨高压氧(HBO)处理肺癌A549细胞对常见肺癌化疗药物敏感性的影响及其作用机制。方法:选取顺铂(Cis)、吉西他滨(Gem)及长春新碱(VCR)这3种临床上常用的肺癌化疗药物作为研究对象。肺癌A549细胞培养完成后,按照实验要求,将细胞分成8组:对照组(不作HBO及药物处理),HBO组(只作0.20 MPa HBO处理3 h),Cis组(只给予20μmol/L Cis处理),HBO+Cis组(HBO预处理3 h后加入20μmol/L Cis),Gem组(只给予20μmol/L Gem处理),HBO+Gem组(HBO预处理3 h后加入20μmol/L Gem),VCR组(只给予20μmol/L VCR处理),HBO+VCR组(HBO预处理3 h后加入20μmol/L VCR)。CCK-8法检测A549细胞增殖情况;流式细胞术检测A549细胞凋亡率及细胞内活性氧(ROS)水平;Western blotting实验检测A549细胞凋亡蛋白的表达。结果:HBO+Cis组、HBO+Gem组和HBO+VCR组A549细胞存活率明显低于Cis组、Gem组和VCR组,差异均有统计学意义(P<0.05)。Cis、Gem和VCR单独处理组A549细胞凋亡比例分别为(44.18±1.27)%、(51.64±2.38)%和(40.38±1.59)%,HBO+Cis组、HBO+Gem组和HBO+VCR组A549细胞凋亡比例分别为(62.37±2.46)%、(75.28±3.59)%和(65.29±1.58)%,差异均有统计学意义(P<0.05)。HBO+Cis组、HBO+Gem组和HBO+VCR组A549细胞中ROS水平明显高于Cis、Gem和VCR单独处理组,差异均有统计学意义(P<0.05)。加入ROS清除剂N-乙酰半胱氨酸(NAC)后,HBO+Cis组、HBO+Gem组和HBO+VCR组A549细胞凋亡被抑制,与未加NAC各组A549细胞的凋亡率比较,差异均有统计学意义(P<0.05)。与Cis组、Gem组和VCR组比较,HBO+Cis组、HBO+Gem组和HBO+VCR组A549细胞凋亡蛋白Bcl-2的表达水平明显降低,caspase-3、PARP和Bax的表达水平明显升高,差异均有统计学意义(P<0.05);与HBO+Cis组、HBO+Gem组和HBO+VCR组比较,Cis+HBO+NAC组、GEM+HBO+NAC组和VCR+HBO+NAC组A549细胞凋亡蛋白Bcl-2的表达水平明显升高,caspase-3、PARP和Bax的表达水平明显降低,差异均有统计学意义(P<0.05)。结论:HBO处理能够增强肺癌A549细胞�Objective To investigate whether hyperbaric oxygen(HBO)can enhance the sensitivity of A549 cells to common chemotherapy drugs for lung cancer and explore its mechanism.Methods The three commonly used chemotherapy drugs for lung cancer,i.e.,cisplatin(Cis),gemcitabine(Gem),and vincristine(VCR),were selected for research.Upon the completion of the cell culture,the A549 cells were divided into eight groups according to the experimental design,including control group(did not receive HBO therapy or chemotherapy drug treatment),HBO group(only given 0.20 MPa HBO for 3 h),Cis group(only given 20μmol/L of Cis),HBO+Cis group(given 20μmol/L of Cis after HBO treatment for 3 h),Gem group(only given 20μmol/L of Gem),HBO+Gem group(given 20μmol/L of Gem after HBO treatment for 3 h),VCR group(only given 20μmol/L of VCR),and HBO+VCR group(given 20μmol/L of VCR after HBO treatment for 3 h).CCK-8 assay was used to detect the proliferation of A549 cells.Flow cytometry was used to detect the apoptosis of A549 cells and the changes of reactive oxygen species(ROS)in the cells.Western blot was used to detect the expression of proteins related to the apoptosis of A549 cells.Results The survival rates of A549 cells in the HBO+Cis,HBO+Gem,and HBO+VCR groups were significantly lower than those in the Cis,Gem,and VCR groups,and all the differences were statistically significant(P<0.05).The A549 cells apoptosis ratios in the Cis,Gem,and VCR groups were(44.18±1.27)%,(51.64±2.38)%,and(40.38±1.59)%,respectively,while the apoptosis ratios in the HBO+Cis,HBO+Gem,HBO+VCR groups were(62.37±2.46)%,(75.28±3.59)%,(65.29±1.58)%,compared with those in the groups without HBO therapy,and all the differences were statistically significant(P<0.05).The levels of ROS in A549 cells in the HBO+Cis,HBO+Gem,and HBO+VCR groups were significantly higher than those in the Cis,Gem,and VCR groups,and the differences were statistically significant(P<0.05).After adding N-acetylcysteine(NAC),a ROS scavenger,the apoptosis of A549 cells in the HBO+Cis,HBO+Gem,and

关 键 词：高压氧 肺癌A549细胞 顺铂 吉西他滨 长春新碱 活性氧 

分 类 号：R734.2[医药卫生—肿瘤]