T-cadherin调控Caspase-1介导的细胞焦亡影响子宫内膜癌体外放射敏感性  

T-cadherin regulates Caspase-1 mediated cell pyrolysis and affects the sensitivity of endometrial cancer to radiotherapy in vitro

作　　者：兰代群 闫志强 林丽慧 陈真 任青 Lan Daiqun;Yan Zhiqiang;Lin Lihui;Chen Zhen;Ren Qing(Department of Obstetrics and Gynecology,Hainan Western Central Hospital,Danzhou 572900,China)

机构地区：[1]海南西部中心医院妇产科,儋州572900

出　　处：《中华放射肿瘤学杂志》2023年第11期1003-1009,共7页Chinese Journal of Radiation Oncology

基　　金：海南省卫生健康行业科研项目(19A200096)。

摘　　要：目的探究T-钙黏蛋白(T-cadherin)是否通过调控胱天蛋白酶1(Caspase-1)介导的细胞焦亡途径影响子宫内膜癌细胞的放射敏感性。方法收集海南西部中心医院2019年10月—2021年3月收治的82例子宫内膜癌患者手术切除的子宫内膜癌组织及癌旁组织标本,免疫组织化学染色和实时反转录PCR(RT-qPCR)检测子宫内膜癌与癌旁组织样本中T-cadherin表达。通过转染pcDNA3.1-T-cadherin慢病毒至人子宫内膜癌细胞系Ishikawa,建立高表达T-cadherin的Ishikawa稳定表达细胞株。Ishikawa细胞给予2 Gy X射线处理,培养24、48、72 h后检测细胞增殖活性。将Ishikawa细胞分为对照组(正常培养的Ishikawa细胞)、照射组(给予2 Gy X射线照射)、pcDNA3.1-NC+照射组(转染pcDNA3.1-NC后给予2 Gy X射线照射)、pcDNA3.1-T-cadherin+照射组(转染pcDNA3.1-T-cadherin后给予2 Gy X射线)、pcDNA3.1-T-cadherin+VA765+照射组(转染pcDNA3.1-T-cadherin,加10μmol/L VA765处理再给予2 Gy X射线照射),进行对应处理后,CCK-8法检测细胞存活率,克隆形成实验检测细胞增殖情况,乳酸脱氢酶(LDH)释放实验检测LDH释放水平,蛋白质印迹法(Western blot)检测Caspase-1、白介素(IL)-1β、IL-18及焦孔素A(gasdermin A,GSDMA)表达,免疫荧光染色检测Caspase-1表达。使用SPSS 23.0统计软件进行数据分析,多组数据比较用单因素方差分析,两两比较采用LSD-t检验。结果子宫内膜癌组织的T-cadherin蛋白阳性表达率为6.09%,低于癌旁组织的87.80%(t=58.48,P<0.01),子宫内膜癌组织中T-cadherin mRNA相对表达量为1.00±0.07,低于较癌旁组织的4.02±0.38(t=32.35,P<0.01)。pcDNA3.1-T-cadherin+照射组细胞活性下降,细胞克隆形成数目减少,LDH释放水平增加,细胞中Caspase-1、IL-1β、IL-18及GSDMA蛋白相对表达量均升高,Caspase-1蛋白红色荧光增强(P<0.01);pcDNA3.1-T-cadherin+VA765+照射组细胞活性升高,LDH释放水平减少,Caspase-1、IL-1β、IL-18及GSDMA蛋白相对表达量均下降,CObjective To investigate whether T-cadherin affects the radiotherapy sensitivity of endometrial cancer cells by regulating the Caspase-1 mediated pyrolysis pathway.Methods Endometrial cancer and adjacent tissue samples were surgically obtained from 82 patients admitted to Hainan Western Central Hospital from October 2019 to March 2021.Immunohistochemical staining and qRT-PCR were used to detect the expression of T-cadherin in endometrial cancer and adjacent tissue samples.By transfecting pcDNA3.1-T-cadherin lentivirus to human endometrial cancer cell line Ishikawa,a stable Ishikawa cell line with high T-cadherin expression was established.Ishikawa cells were treated with 2 Gy X-ray,and the cell proliferation activity was detected after 24,48,72 h of culture.The cells were divided into the control group(normally cultured Ishikawa cells),irradiation group(treated with 2 Gy X-ray irradiation),pcDNA3.1-NC+irradiation group(transfected with pcDNA3.1-NC followed by 2 Gy X-ray irradiation),pcDNA3.1-T-cadherin+irradiation group(transfected with pcDNA3.1-T-cadherin followed by 2 Gy X-ray irradiation),pcDNA3.1-T-cadherin+VA765+irradiation group(transfected with pcDNA3.1-T-cadherin,plus 10μmol/L VA765 treatment followed by 2 Gy X-ray irradiation).After corresponding treatment,cell survival rate was detected by CCK-8 assay.Cell proliferation was determined by colony formation assay.The level of lactate dehydrogenase(LDH)release was measured by LDH release test.The expression levels of Caspase-1,interleukin(IL)-1β,IL-18 and gasdermin A(GSDMA)were detected by Western blot.The expression level of Caspase-1 was detected by immunofluorescence staining.SPSS 23.0 statistical software was used for data analysis.One-way ANOVA was used for data comparison among multiple groups.LSD-t test was used for two-paired comparison.Results The positive expression rate of T-cadherin protein in endometrial cancer tissues was 6.09%,lower than 87.80%in adjacent normal tissues(t=58.48,P<0.01).The relative expression level of T-cadherin mRNA in end

关 键 词：子宫内膜肿瘤 T-钙黏蛋白 细胞焦亡 放射敏感性 

分 类 号：R737.33[医药卫生—肿瘤]