GDF11调控NLRP3-焦亡途径抑制宫颈癌增殖和转移的作用机制研究  

Study on the mechanism of growth differentiation factor 11 inhibiting proliferation and metastasis of cervical cancer by regulating nucleotide-binding oligomerization domain-like receptors 3-pyroptosis pathway

作　　者：李帆 施兴华 朱维培[3] LI Fan;SHI Xinghua;ZHU Weipei(Department of Gynecology,Shanghai Xuhui Central Hospital(Zhongshan-Xuhui Hospital,Fudan University),Shanghai 200030,China;Department of Gynecology,Qidong Maternal and Child Health Hospital(Qidong Branch,Pediatrics Hospital Affiliated to Fudan University),Qidong 226200,Jiangsu,China;Department of Gynecology,the Second Affiliated Hospital of Soochow University,Suzhou 215004,Jiangsu,China)

机构地区：[1]上海市徐汇区中心医院(复旦大学附属中山医院徐汇医院)妇科,上海200030 [2]启东市妇幼保健院(复旦大学附属儿科医院启东分院)妇科,江苏启东226200 [3]苏州大学附属第二医院妇科,江苏苏州215004

出　　处：《中国性科学》2023年第10期69-73,共5页Chinese Journal of Human Sexuality

摘　　要：目的探讨生长分化因子11(GDF11)对宫颈癌细胞增殖、转移的影响及其作用机制分析。方法采用实时荧光定量聚合酶链反应(qRT-PCR)检测人正常宫颈上皮细胞End1/E6E7和宫颈癌细胞系HeLa、C33A、SiHa、Caski中GDF11、NOD样受体家族蛋白3(NLRP3)表达情况,构建GDF11过表达慢病毒载体、NLRP3过表达慢病毒载体,转染/共转染HeLa细胞系,分别记为oe-GDF11组和oe-GDF11/oe-NLRP3组,同时设置Control组和vector组。采用qRT-PCR、酶联免疫吸附试验(ELISA)法检测HeLa细胞中白介素(IL)-1β、IL-18 mRNA水平和含量,采用CCK8法、克隆形成试验、划痕试验、Transwell试验分别检测HeLa细胞活性、增殖、迁移、侵袭情况,WB法检测基质金属蛋白酶(MMPs)和上皮间质转化(EMT)相关蛋白表达水平。结果HeLa、C33A、SiHa、Caski细胞中GDF11 mRNA水平均低于End1/E6E7细胞(P<0.05),而NLRP3 mRNA水平均高于End1/E6E7细胞(P<0.05)。oe-GDF11组NLRP3 mRNA水平(0.41±0.05)低于Control组(1.03±0.05)(P<0.05)。e-GDF11组细胞IL-1β、IL-18 mRNA水平和含量、细胞活力、克隆形成率、划痕闭合率、单个视野细胞侵袭数,以及MMP2、MMP9、N-cadherin、Vimentin蛋白表达均低于Control组(P<0.05),E-cadherin蛋白表达高于Control组(P<0.05)。oe-GDF11/oe-NLRP3组细胞IL-1β、IL-18 mRNA水平和含量、细胞活力、克隆形成率、划痕闭合率、单个视野细胞侵袭数,以及MMP2、MMP9、N-cadherin、Vimentin蛋白表达均高于oe-GDF11组(P<0.05),E-cadherin蛋白表达低于oe-GDF11组(P<0.05)。结论GDF11过表达可抑制宫颈癌细胞增殖和转移,可能与抑制NLRP3-焦亡途径有关。Objective To explore the effects and action mechanism of growth differentiation factor 11(GDF11)on the proliferation and metastasis of cervical cancer cells.Methods The expressions of GDF11 and nucleotide-binding oligomerization domain-like receptors 3(NLRP3)in human normal cervical epithelial cells End1/E6E7 and cervical cancer cell lines HeLa,C33A,SiHa,Caski were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The lentivirus vectors with GDF11 and NLRP3 overexpression were constructed,and HeLa cell lines were transfected/co-transfected with them,which were recorded as oe-GDF11 group and oe-GDF11/oe-NLRP3 group.Control group and vector group were set up at the same time.The mRNA levels and content of interleukin(IL)-1βand IL-18 in HeLa cells were detected by qRT-PCR and enzyme-linked immunosorbent assay(ELISA).The activity,proliferation,migration and invasion of HeLa cells were detected by Cell Counting Kit-8(CCK8),clone formation assay,scratch assay and Transwell assay.The expression levels of matrix metalloproteinases(MMPs)and epithelial-mesenchymal transition(EMT)related proteins were detected by western blotting(WB).Results The mRNA level of GDF11 in HeLa,C33A,SiHa and Caski cells was lower than that in End1/E6E7 cells(P<0.05),while mRNA level of NLRP3 was higher than that in End1/E6E7 cells(P<0.05).The mRNA level of NLRP3 in oeGDF11 group was lower than that in control group[(0.41±0.05)vs.(1.03±0.05),P<0.05].The mRNA levels and content of IL-1βand IL-18,cells activity,clone formation rate,scratch closure rate,number of invasion cells in single field and expressions of MMP2,MMP9,N-cadherin and Vimentin proteins in eGDF11 group were lower than those in control group(P<0.05),while expression of E-cadherin protein was higher than that in control group(P<0.05).The mRNA levels and content of IL-1βand IL-18,cells activity,clone formation rate,scratch closure rate,number of invasion cells in single field and expressions of MMP2,MMP9,N-cadherin and Vimentin proteins in oe-GDF1

关 键 词：宫颈癌 生长分化因子11 NOD样受体家族蛋白3 细胞焦亡 细胞增殖 转移 

分 类 号：R711[医药卫生—妇产科学]