新型精胺氧化酶抑制药对人胃癌SGC-7901细胞增殖、凋亡和自噬的影响  

Effects of a novel spermine oxidase inhibitor on cell proliferation,apoptosis,autophagy of human gastric cancer SGC-7901 cells

作　　者：李玉玲 杨建林[1] 崔芝 王静[1] 吴红艳[1] 吕亚丰 曹春雨[1] LI Yu-ling;YANG Jian-lin;CUI Zhi;WANG Jing;WU Hong-yan;LÜYa-feng;CAO Chun-yu(Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,Medical College of China Three Gorges University,Yichang 443002,Hubei Province,China)

机构地区：[1]三峡大学基础医学院肿瘤微环境与免疫治疗湖北省重点实验室,湖北宜昌443002

出　　处：《中国临床药理学杂志》2023年第16期2343-2347,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81372265,30772590);湖北省卫生健康科研基金资助项目(WJ2019H532);肿瘤微环境与免疫治疗湖北省重点实验室开放基金资助项目(2019KZL01,2016KZL04)。

摘　　要：目的研究新型精胺氧化酶抑制药SI-4650对人胃癌SGC-7901细胞增殖、凋亡和自噬的影响。方法将SGC-7901细胞分为空白组(正常培养)、低、高剂量实验组(40、80μmol·L^(-1)SI-4650)、自噬抑制药3-甲基腺嘌呤(3-MA)组(2.5 mmol·L^(-1)3-MA)、3-MA+低、高剂量实验组(2.5 mmol·L^(-1)3-MA+40、80μmol·L^(-1)SI-4650),均处理48 h。用化学发光法检测细胞中精胺氧化酶(SMO)活性,用细胞计数8(CCK8)法和流式细胞术检测细胞增殖和周期,用碘化丙啶(PI)/异硫氰酸荧光素(FITC)-Annexin V双染法检测凋亡细胞情况,用蛋白质印迹法检测自噬相关蛋白微管结合蛋白轻链-3Ⅰ/Ⅱ(LC3-Ⅰ/Ⅱ)表达水平。结果空白组与低、高剂量实验组中人胃癌SGC-7901细胞中相对SMO酶活性分别为(7293±195)、(4506±195)、(989±115)RLU·(mg·s)^(-1),S期细胞比例分别为(28.83±1.17)%、(32.23±1.21)%、(36.50±0.73)%,低、高剂量实验组与空白组相比,差异均有统计学意义(均P<0.01)。空白组与低、高剂量实验组以及3-MA组与3-MA+低剂量实验组、3-MA+高剂量实验组中可见,细胞生长抑制率分别为(0.00±2.09)%、(43.61±2.29)%、(52.16±2.49)%、(1.81±4.43)%、(27.50±1.88)%、(34.22±1.07)%,细胞凋亡率分别为(7.01±2.09)%、(13.16±1.59)%、(25.35±2.32)%、(7.13±2.09)%、(8.61±1.59)%、(11.35±2.32)%,自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值分别为0.01±0.03、0.58±0.04、2.73±0.03、0.03±0.01、0.06±0.02、0.23±0.03。低、高剂量实验组与空白组相比,低、高剂量实验组与对应浓度3-MA+低、高剂量实验组相比,差异均有统计学意义(均P<0.01)。结论SI-4650能高效抑制人胃癌SGC-7901细胞增殖,机制可能与干扰多胺代谢、阻滞细胞周期S期和诱导细胞自噬、凋亡相关。Objective To investigate the effects a novel spermine oxidase(SMO)inhibitor SI-4650 on cell proliferation,apoptosis,autophagy of human gastric cancer SGC-7901 cells.Methods SGC-7901 cells were divided into blank control group(normal culture),experimental-L,-H groups(40,80μmol·L^(-1) SI-4650),autophagy inhibitor 3-methyladenine(3-MA)group(2.5 mmol·L^(-1)3-MA),and 3-MA+experimental-L,-H groups(2.5 mmol·L^(-1)3-MA+40,80μmol·L^(-1) SI-4650).Chemiluminescence was used to analyze spermine oxidase(SMO)activity;the CCK8 method and flow cytometry were used to detect the inhibitory effect on proliferation and cell cycle;PI/FITC-Annexin V double staining analysis was used to detect the apoptosis;Western blot was used to analyze the expression levels of autophay-related protein(LC3-Ⅰ/Ⅱ).Results The spermine oxidase enzymatic activity of SGC-7901 cells in blank control group and experimental-L,-H groups were(7293±195),(4506±195),(989±115)RLU·(mg·s)^(-1),and the number of cells in S-phase were(28.83±1.17)%,(32.23±1.21)%,(36.50±0.73)%.Comparison between blank control group and experimental groups showed all the differences were significant(all P<0.01).It could be seen in blank group,experimental-L,-H groups,3-MA group,3-MA+experimental-L,-H groups that the Growth inhibition rates were(0.00±2.09)%,(43.61±2.29)%,(52.16±2.49)%,(1.81±4.43)%,(27.50±1.88)%,(34.22±1.07)%;proportions of apoptotic cells were(7.01±2.09)%,(13.16±1.59)%,(25.35±2.32)%,(7.13±2.09)%,(8.61±1.59)%,(11.35±2.32)%;values of LC3-Ⅱ/LC3-Ⅰwere 0.01±0.03,0.58±0.04,2.73±0.03,0.03±0.01,0.06±0.02,0.23±0.03.Comparisons of blank control group with experimental-L,-H groups,experimental-L,-H groups with the corresponding concentration of 3-MA+experimental-L,-H groups showed that the differences were significant(all P<0.01).Conclusion SI-4650 has the pharmacological activity of killing human gastric cancer SGC-7901,and its mechanism may be related to the interference of polyamine metabolism and induction of cell apoptosis and autophagy.

关 键 词：精胺氧化酶抑制药 人胃癌SGC-7901细胞 增殖 凋亡 自噬 

分 类 号：R97[医药卫生—药品]