双氢青蒿素、焦孔素E对喉癌细胞增殖、转移及焦亡的影响  

Effect of dihydroartemisinin and gasdermin E on the proliferation,migration,and pyroptosis of laryngeal cancer cells

作　　者：金玲爽[1] 郭慧娜 杨立[3] 高泽慧 金胜勇 王晶[6] Jin Lingshuang;Guo Huina;Yang Li;Gao Zehui;Jin Shengyong;Wang Jing(Department of Head and Neck Surgery,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China;Department of Otolaryngology Head and Neck Tumor,Shanxi Provincial Key Laboratory,First Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Ultrasound,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China;Department of Otolaryngology Head and Neck Surgery,First Hospital of Shanxi Medical University,Taiyuan 030001,China;Physical Examination Center,Traditional Chinese Hospital of Funing County,Qinhuangdao 066300,China;Department of Pathology,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China)

机构地区：[1]山西省肿瘤医院,中国医学科学院肿瘤医院,山西医院山西医科大学附属肿瘤医院头颈外科,太原030013 [2]山西医科大学第一医院耳鼻咽喉头颈肿瘤山西省重点实验室,太原030001 [3]山西省肿瘤医院,中国医学科学院肿瘤医院山西医院,山西医科大学附属肿瘤医院超声科,太原030013 [4]山西医科大学第一医院耳鼻咽喉头颈外科,太原030001 [5]秦皇岛市抚宁县中医院体检中心,秦皇岛066300 [6]山西省肿瘤医院,中国医学科学院肿瘤医院山西医院,山西医科大学附属肿瘤医院病理科,太原030013

出　　处：《肿瘤研究与临床》2024年第8期615-621,共7页Cancer Research and Clinic

摘　　要：目的探讨双氢青蒿素(DHA)、焦孔素E(GSDME)对喉癌细胞增殖、转移及焦亡的影响及相关机制。方法取人喉鳞状细胞癌Hep-2细胞,将其分为空白组(未干预的Hep-2细胞)、DHA组(采用50μmol/L DHA处理的Hep-2细胞)、GSDME-siRNA组(转染GSDME-siRNA的Hep-2细胞)、DHA+GSDME-siRNA组(采用50μmol/L DHA培养并转染GSDME-siRNA的Hep-2细胞)。采用四甲基偶氮唑盐(MTT)法检测DHA对Hep-2细胞增殖能力的影响,计算细胞增殖抑制率及半数抑制浓度(IC 50);流式细胞术检测细胞焦亡率;Transwell法检测细胞侵袭能力;蛋白质印迹法检测焦亡相关蛋白GSDME、caspase-3及己糖激酶Ⅱ(HK-Ⅱ)、亲环素D、电压依赖性阴离子通路(VDAC)蛋白的相对表达水平。结果10、20、40、80、160μmol/L DHA作用Hep-2细胞48 h的细胞增殖抑制率均高于相应浓度作用24 h的细胞增殖抑制率(均P<0.05);DHA作用Hep-2细胞24、48 h的IC 50值分别为57.20、43.50μmol/L,故选择50μmol/L DHA进行后续实验。空白组、DHA组、GSDME-siRNA组及DHA+GSDME-siRNA组细胞焦亡率分别为(6.5±0.8)%、(22.7±2.5)%、(3.1±0.6)%及(7.0±1.0)%,差异有统计学意义(F=221.20,P<0.05);细胞侵袭数分别为(153±14)个、(95±10)个、(205±16)个及(148±16)个,差异有统计学意义(F=56.89,P<0.05)。蛋白质印迹法检测结果显示,DHA组GSDME、caspase-3相对表达水平均高于空白组(均P<0.05);GSDME-siRNA组GSDME、caspase-3相对表达水平均低于DHA组(均P<0.05);DHA+GSDME-siRNA组GSDME、caspase-3相对表达水平均高于GSDME-siRNA组(均P<0.05);DHA组HK-Ⅱ、亲环素D、VDAC相对表达水平均低于空白组(均P<0.05);GSDME-siRNA组HK-Ⅱ、亲环素D、VDAC相对表达水平均高于DHA组(均P<0.05);DHA+GSDME-siRNA组HK-Ⅱ、亲环素D、VDAC相对表达水平均低于GSDME-siRNA组(均P<0.05)。结论双氢青蒿素可增加喉癌细胞焦亡,降低细胞增殖及转移能力,可能与抑制线粒体HK-Ⅱ表达相关。ObjectiveTo investigate the effect of dihydroartemisinin(DHA)and gasdermin E(GSDME)on the proliferation,metastasis and pyroptosis of laryngeal cancer cells as well as its related mechanisms.MethodsHuman laryngeal squamous cell cancer Hep-2 cells were taken and divided into 4 groups:the blank group(untreated Hep-2 cells),DHA group(Hep-2 cells treated with 50μmol/L DHA),GSDME-siRNA group(Hep-2 cells transfected with GSDME-siRNA),and DHA+GSDME-siRNA group(Hep-2 cells treated with 50μmol/L DHA and transfected with GSDME-siRNA).Methyl thiazolyl tetrazolium(MTT)method was used to detect the effect of DHA on the proliferation ability of Hep-2 cells,and the cell proliferation inhibition rate and half inhibitory concentration(IC 50)were calculated.Flow cytometry was used to detect the pyroptosis rate,Transwell assay was used to detect cell invasion ability and Western blot was used to detect the relative expression levels of GSDME,caspase-3,hexokinaseⅡ(HK-Ⅱ),cyclophilin D,and voltage-dependent anion channel(VDAC)proteins.ResultsThe cell proliferation inhibition rates of Hep-2 cells treated with 10,20,40,80,160μmol/L DHA for 48 h were higher than those treated with the corresponding concentration for 24 h(all P<0.05).The IC 50 values of Hep-2 cells treated by DHA for 24 h and 48 h were 57.20μmol/L and 43.50μmol/L,respectively,and thus 50μmol/L DHA was selected for subsequent experiments.The pyroptosis rate was(6.5±0.8)%,(22.7±2.5)%,(3.1±0.6)%and(7.0±1.0)%,respectively in the blank group,DHA group,GSDME-siRNA group,and DHA+GSDME-siRNA group,and the difference was statistically significant(F=221.20,P<0.05).The number of invasive cells was(153±14),(95±10),(205±16),and(148±16),respectively in the blank group,DHA group,GSDME-siRNA group,and DHA+GSDME-siRNA group,and the difference was statistically significant(F=56.89,P<0.05).The results of Western blot showed that the relative expression levels of GSDME and caspase-3 in DHA group were higher than those in the blank group(both P<0.05);the relative expression l

关 键 词：喉肿瘤 青蒿素类 己糖激酶 焦孔素E 

分 类 号：R739.65[医药卫生—肿瘤]