硒对顺铂诱导的肺癌细胞增殖、凋亡、迁移、侵袭及MDA、GSH-Px活性的影响  

Effects of selenium on the proliferation, apoptosis, migration, invasion, MDA and GSH Px activities of lung cancer cells induced by cisplatin

作　　者：丁华 郭浩 陈杉 王淼 龙芬 陈坤 冉瑞智 Ding Hua;Guo Hao;Chen Shan;Wang Miao;Long Fen;Chen Kun;Ran Ruizhi(Department of Oncology,Enshizhou Tujia and Miao Autonomous Prefecture Central Hospital,Hubei Province,Enshi 445000,China)

机构地区：[1]湖北省恩施土家族苗族自治州中心医院肿瘤二科,445000

出　　处：《疑难病杂志》2023年第1期30-35,共6页Chinese Journal of Difficult and Complicated Cases

基　　金：湖北省卫生健康委员会资助项目(WJ2021M259)。

摘　　要：目的探讨硒对顺铂(CIS)诱导的肺癌细胞(A549)增殖、凋亡、迁移、侵袭及丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)活性的影响。方法2021年7月—2022年3月于恩施州土家族苗族自治州中心医院硒实验室进行实验。以对数期的A549细胞为研究对象,计算CIS和亚硒酸钠(Na2SeO3)的半数抑制浓度(IC50)。将A549细胞分为对照组(C组)、CIS组、Na_(2)SeO_(3)组、CIS+Na_(2)SeO_(3)组,C组常规培养48 h,CIS组加入12μmol/L CIS培养48 h,Na_(2)SeO_(3)组加入Na_(2)SeO_(3)110 nmol/L培养48 h,CIS+Na_(2)SeO_(3)组先加入Na_(2)SeO_(3)110 nmol/L培养24 h,随后加入12μmol/L CIS继续培养24 h。采用CCK-8法检测A549细胞活力,流式细胞仪检测A549细胞凋亡,Western-blot检测凋亡、迁移侵袭蛋白表达,Transwell实验检测A549细胞迁移、侵袭个数;测定细胞活性氧(ROS)、MDA、4-羟基壬烯醛(4-HNE)、GSH-Px水平。结果CIS的IC50为12.02μmol/L,Na_(2)SeO_(3)的IC_(50)为110.20 nmol/L。与C组比较,CIS组、Na_(2)SeO_(3)组A549细胞活力、迁移个数、侵袭个数、MMP-9、MMP-3蛋白水平均显著降低(F/P=147.876/<0.001、78.926/<0.001、43.071/<0.001、36.862/<0.001、75.431/<0.001),凋亡率、cleaved-caspase9、cleaved-caspase3蛋白、ROS、MDA、4-HNE水平显著升高(F/P=15.625/<0.001、8.131/0.004、22.371/<0.001、45.779/<0.001、5.216/0.019、25.084/<0.001),GSH-Px活性比较差异无统计学意义(P>0.05)。与CIS组比较,CIS+Na_(2)SeO_(3)组A549细胞活力、迁移个数、侵袭个数、MMP-9、MMP-3蛋白水平降低(t/P=9.340/<0.001、9.573/<0.001、9.746/<0.001、6.788/0.001、9.481/<0.001),凋亡率、cleaved-caspase9、cleaved-caspase3蛋白、ROS、MDA、4-HNE水平升高(t/P=10.911/<0.001、9.098/<0.001、10.809/<0.001、6.069/0.002、9.218/<0.001、15.945/<0.001),GSH-Px活性比较差异无统计学意义(P>0.05)。结论Na_(2)SeO_(3)可加强CIS诱导的A549细胞凋亡,抑制其增殖、迁移、侵袭,可能与促进ROS、MDA产生有关。Objective To investigate the effects of selenium on the proliferation, apoptosis, migration, invasion, malondialdehyde(MDA) and glutathione peroxidase(GSH-Px) activities of lung cancer cells(A549) induced by cisplatin(CIS). Methods From July 2021 to March 2022, the experiment was conducted in the Selenium Laboratory of the Central Hospital of Enshi Tujia and Miao Autonomous Prefecture. The median inhibitory concentration(IC50) of CIS and sodium selenite(Na_(2)SeO_(3)) was calculated on A549 cells in logarithmic phase. A549 cells were divided into control group(Group C), CIS group, Na_(2)SeO_(3)group, CIS+Na_(2)SeO_(3)group. Group C was routinely cultured for 48 h, and CIS group was added with 12 μmol/L CIS was cultured for 48 h, Na_(2)SeO_(3)group was added with Na_(2)SeO_(3)110 nmol/L for 48 h, CIS+Na_(2)SeO_(3)group was added with Na_(2)SeO_(3)110 nmol/L for 24 h, and then 12 μmol/L CIS was cultured for 24 h. CCK-8 method was used to detect the viability of A549 cells, flow cytometry was used to detect the apoptosis of A549 cells, Western blot was used to detect the expression of apoptosis and migration invasion proteins, and Transwell test was used to detect the number of A549 cells migrating and invading;The levels of ROS, MDA, 4-HNE and GSH-Px were measured. Results The IC50of CIS is 12.02 μmol/L, IC50of Na_(2)SeO_(3)is 110.20 nmol/L. Compared with group C, the activity, migration, invasion, MMP-9, MMP-3 protein levels of A549 cells in CIS group and Na_(2)SeO_(3)group decreased(F/P=147.876/<0.001, 78.926/<0.001, 43.071/<0.001, 36.862/<0.001, 75.431/<0.001), apoptosis rate, cleaved caspase9, cleaved caspase3 protein, ROS, MDA The level of 4-HNE increased significantly(F/P=15.625/<0.001, 8.131/0.004, 22.371/< 0.001, 45.779/< 0.001, 5.216/0.019, 25.084/< 0.001), and there was no significant difference in GSH-Px activity(P>0.05). Compared with CIS group, A549 cells in CIS+Na_(2)SeO_(3)group had lower activity, migration, invasion, MMP-9, MMP-3 protein levels(t/P=9.340/< 0.001, 9.573/< 0.001, 9.746/<0.001, 6.7

关 键 词：肺癌 亚硒酸钠 顺铂 增殖 迁移 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]