摘要
目的探讨汉防己甲素(tetrandrine,TET)对口腔鳞癌(oral squamous cell carcinoma,OSCC)细胞侵袭潜能的影响及其相关机制。方法将OSCC细胞随机分为4组:对照组(不加药物处理)、低剂量组(10μmol/L TET)、中剂量组(20μmol/L TET)、高剂量组(30μmol/L TET)。采用MTT法、划痕实验和Transwell实验分别测定各组细胞的活力、迁移及侵袭能力。对OSCC细胞分别转染缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、程序性细胞死亡蛋白1(programmed cell death protein 1,PD-1)及程序性细胞死亡蛋白1配体(programmed cell death ligand 1,PD-L1)模拟物及抑制剂,均连续转染24 h。采用RT-qPCR检测正常口腔上皮细胞、OSCC细胞以及转染HIF-1α、PD-1、PD-L1模拟物及抑制剂OSCC细胞中HIF-1α、PD-1、PD-L1 mRNA表达水平。通过Western blot检测各组细胞中纤维连接蛋白(Fibronectin)、波形蛋白(Vimentin)、E-钙黏蛋白(E-cadherin)、HIF-1α、PD-1以及PD-L1蛋白表达水平。结果与对照组相比,低、中、高剂量组细胞活力、迁移及侵袭能力均被显著抑制(P<0.05),HIF-1α、PD-1、PD-L1 mRNA显著上调(P<0.05),Fibronectin、Vimentin、HIF-1α、PD-1、PD-L1蛋白水平随剂量增加逐渐下降(P<0.05),E-cadherin蛋白水平随剂量增加逐渐上升(P<0.05)。转染HIF-1α模拟物细胞的HIF-1α相对表达量及穿膜细胞数高于转染HIF-1α抑制剂细胞(P<0.05),转染PD-1模拟物细胞的PD-1相对表达量及穿膜细胞数高于转染PD-1抑制剂细胞(P<0.05),转染PD-L1模拟物细胞的PD-L1相对表达量及穿膜细胞数高于转染PD-L1抑制剂细胞(P<0.05)。结论TET可能通过抑制HIF-1α/PD-1/PD-L1轴,从而抑制OSCC细胞活力、迁移及侵袭能力,有望作为辅助治疗OSCC的新药。
Objective To explore the effects of tetrandrine(TET)on the invasion potential of oral squamous cell carcinoma(OSCC)cells and its related mechanism.Methods OSCC cells were randomized into control(no drug treatment),low-dose(10μmol/L TET),medium-dose(20μmol/L TET),and high-dose(30μmol/L TET)groups.The cell viability,migration,and invasion abilities in each group were determined by MTT,scratch test,and Transwell assay separately.OSCC cells were respectively transfected with hypoxia-inducible factor-1α(HIF-1α),programmed cell death protein 1(PD-1),and programmed cell death ligand 1(PD-L1)mimics and inhibitors for 24 h continuously.RT-qPCR was used to determine the mRNA expression levels of HIF-1α,PD-1,and PD-L1 in normal oral epithelial cells,OSCC cells,and OSCC cells transfected with HIF-1α,PD-1,and PD-L1 mimics and inhibitors.Western blot was used to check the protein expression levels of Fibronectin,Vimentin,E-cadherin,HIF-1α,PD-1,and PD-L1 in cells of each group.Results Compared with control group,the cell viability,migration,and invasion abilities were significantly inhibited in low-dose,medium-dose,and high-dose groups(P<0.05),which showed an obvious up-regulation in mRNA levels of HIF-1α,PD-1,and PD-L1(P<0.05).The protein levels of Fibronectin,Vimentin,HIF-1α,PD-1,and PD-L1 in low-dose,medium-dose and high-dose groups gradually decreased with the increased dose(P<0.05),while the protein level of E-cadherin was gradually increased(P<0.05).The relative expression level of HIF-1αand the number of transmembrane cells in cells transfected with HIF-1αmimic were higher than those in cells transfected with HIF-1αinhibitor(P<0.05);the relative expression level of PD-1 and the number of transmembrane cells in cells transfected with PD-1 mimic were higher than those in cells transfected with PD-1 inhibitor(P<0.05);and the relative expression level of PD-L1 and the number of transmembrane cells in cells transfected with PD-L1 mimic were higher than those in cells transfected with PD-L1 inhibitor(P<0.05).Concl
作者
彭坷平
田桂湘
彭书旺
宾东华
左巧娟
谭焱
谢海清
PENG Keping;TIAN Guixiang;PENG Shuwang;BIN Donghua;ZUO Qiaojuan;TAN Yan;XIE Haiqing(The First Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China;The Second Xiangya Hospital of Central South University,Changsha,Hunan 410011,China)
出处
《湖南中医药大学学报》
CAS
2023年第11期1971-1977,共7页
Journal of Hunan University of Chinese Medicine
基金
湖南省自然科学基金青年项目(2021JJ40869)
湖南省卫生健康委科研课题项目(202207012427,202206032424)。
