氯胍介导氧化还原驱动的铁死亡诱导膀胱癌细胞凋亡的研究  

Proguanil induces bladder cancer cell apoptosis through mediating oxidation-reduction driven ferroptosis

作　　者：潘清华 刘印龙 刘勇 廖宝春 胡健 朱志坚 PAN Qing-hua;LIU Yin-long;LIU Yong;LIAO Bao-chun;HU Jian;ZHU Zhi-jian(Abdominal Pelvic Oncology,Ganzhou Cancer Hospital(Affiliated Cancer Hospital of Gannan Medical University),Ganzhou 341000,Jiangxi Province,China;Abdominal Surgery,Ganzhou Cancer Hospital(Affiliated Cancer Hospital of Gannan Medical University),Ganzhou 341000,Jiangxi Province,China;Departmentof Pharmacy,Ganzhou Cancer Hospital(Affiliated Cancer Hospital of Gannan Medical University),Ganzhou 341000,Jiangxi Province,China)

机构地区：[1]赣州市肿瘤医院(赣南医科大学附属肿瘤医院)腹盆部肿瘤内科,江西赣州341000 [2]赣州市肿瘤医院(赣南医科大学附属肿瘤医院)腹部外科,江西赣州341000 [3]赣州市肿瘤医院(赣南医科大学附属肿瘤医院)药学部,江西赣州341000

出　　处：《中国临床药理学杂志》2024年第20期2988-2992,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨氯胍对膀胱癌细胞增殖、凋亡影响的潜在作用机制。方法将253J细胞随机分为对照组(正常培养)、氯胍组(42.06μmol·L^(-1)氯胍)、pcDNA组(转染pcDNA+42.06μmol·L^(-1)氯胍)、FADS2组[转染脂肪酸去饱和酶基因2(FADS2)+42.06μmol·L^(-1)氯胍]、si-NC组(转染si-NC)、si-FADS2组(转染si-FADS2)、Ferrostatin-1组(转染si-FADS2+10μmol·L^(-1)铁抑素-1)。用实时荧光定量聚合酶链反应(RT-qPCR)实验检测相关基因相对表达水平,用蛋白质印迹实验检测各蛋白相对表达水平,用原位末端转移酶标记法(Tunel)实验检测细胞凋亡情况,用5-乙炔基-2’-脱氧尿苷(EdU)实验检测细胞增殖情况,用Transwell实验检测细胞迁移能力,用试剂盒法测定Fe^(2+)水平,用DCFH-DA探针检测活性氧(ROS)水平。结果对照组、氯胍组、pcDNA组、FADS2组细胞FADS2 mRNA水平分别为1.00±0.11、0.47±0.09、0.49±0.06和2.09±0.21,细胞增殖率分别为(100.00±3.50)%、(54.31±4.90)%、(56.46±5.17)%和(78.76±6.50)%,凋亡率分别为(3.92±0.53)%、(28.79±3.30)%、(27.20±2.90)%和(7.34±0.68)%,细胞迁移数分别为(132.70±9.81)、(70.10±5.05)、(68.70±5.37)和(101.80±11.25)个,Fe^(2+)水平分别为(100.00±8.14)%、(201.33±17.84)%、(192.38±21.34)%和(116.70±10.90)%,谷胱甘肽过氧化物酶4(GPX4)蛋白相对表达水平分别为0.77±0.05、0.31±0.05、0.34±0.05和0.68±0.06。氯胍组的上述指标分别与对照组比较,FADS2组的上述指标分别与pcDNA组比较,在统计学上差异均有统计学意义(均P<0.05)。si-NC组、si-FADS2组、Ferrostatin-1组ROS水平分别为9.72±1.18、40.94±5.63和13.77±1.40。si-FADS2组与si-NC组比较,Ferrostatin-1组与si-FADS2比较,在统计学上差异均有统计学意义(均P<0.05)。结论氯胍可通过抑制FADS2表达介导氧化还原驱动的铁死亡途径诱导膀胱癌细胞凋亡。Objective To explore the potential mechanism of proguanil on the proliferation and apoptosis of bladder cancer cells.Methods253J cells were randomly divided into control group(normal treatment),proguanil group(42.06μmol·L^(-1)proguanil),pcDNA group(transfected with pcDNA+42.06μmol·L^(-1)proguanil),FADS2 group[transfected fatty acid desaturase gene 2(FADS2)+42.06μmol·L^(-1)proguanil],si-NC(transfection si-NC),si-FADS2(transfection si-FADS2),Ferrostatin-1 group(transfected with si-FADS2+10μmol·L^(-1)ferrostatin-1).Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)assay was used to detect mRNA expression of related genes;Western blot assay was used to detect the expression of each protein;apoptosis was detected by TdT mediated dUDP nick end labeling(Tunel)assay;5-ethynyl-2'-deoxyuridine(EdU)assay to detect cell proliferation;the Transwell assay measures the ability of cells to migrate;Fe^(2+)levels were determined by kit method;DCFH-DA probe was used to detect ROS levels.Results The mRNA levels of FADS2 in control group,proguanil group,pcDNA group and FADS2 group were 1.00±0.11,0.47±0.09,0.49±0.06 and 2.09±0.21,respectively;cell proliferation rate were(100.00±3.50)%,(54.31±4.90)%,(56.46±5.17)%and(78.76±6.50)%,respectively;the apoptosis rate were(3.92±0.53)%,(28.79±3.30)%,(27.20±2.90)%and(7.34±0.68)%,respectively;the migration number were132.70±9.81,70.10±5.05,68.70±537 and 101.80±11.25,respectively;Fe^(2+)level were(100.00±8.14)%,(201.33±17.84)%,(192.38±21.34)%and(116.70±10.90)%,respectively;GPX4 protein relative expression level were 0.77±0.05,0.31±0.05,0.34±0.05 and 0.68±0.06,respectively.The above indexes in proguanil group were compared with those in control group,the above indexes in FADS2 group were compared with those in pcDNA group,all the differences were statistically significant(all P<0.05).The ROS levels of si-NC,si-FADS2 and Ferrostatin-1 groups were 9.72±1.18,40.94±5.63 and 13.77±1.40,respectively.Compared the si-FADS2 group with the si-NC group,Fe

关 键 词：氯胍 膀胱癌 脂肪酸去饱和酶基因2 铁死亡 凋亡 

分 类 号：R979.1[医药卫生—药品]