PPAR-γ激动剂吡格列酮抑制炎症介质HMGB1对恶性间皮瘤细胞增殖活力的影响  

Effect of PPAR-γ agonist pioglitazone on the prolifeiration of malignant nesothelionma cells induced by HMGB1

作　　者：王彦镔 沈蔚 干译涵 邹瑾 张宇 朱丽瑾 鞠莉 蒋兆强 应士波 Wang Yanbin;Shen Wei;Gan Yihan;Zou Jin;Zhang Yu;Zhu Lijin;Ju Li;Jiang Zhaoqiang;Ying Shibo(Hangzhou Medical College,Hangzhou 310013,China;Department of Respiratory Medicine,The Third People's Hospital of Cixi,Ningbo 315324,China)

机构地区：[1]杭州医学院,310013 [2]慈溪市第三人民医院呼吸内科,宁波315324

出　　处：《中华劳动卫生职业病杂志》2021年第9期641-647,共7页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金项目(81973011);慈溪市科技计划项目(CN2018023、CN2019031)。

摘　　要：目的探讨PPAR-γ激动剂吡格列酮(piglitazone,PGZ)对恶性间皮瘤细胞增殖活力的影响及其作用机制。方法于2019年12月,选择人双相型恶性间皮瘤细胞株(MSTO-211H)和人上皮型恶性间皮瘤细胞株(NCI-H2452),给予不同终浓度的PGZ(0、10、50、100、150、200μmol/L)处理24、48、72 h后,采用CCK8法检测细胞增殖活力,在倒置显微镜下观察细胞数量及形态的变化,实时荧光定量反转录聚合酶链反应(qRT-PCR)检测0、10、50、100μmol/L PGZ处理细胞72 h时,细胞内PPAR-γ、高迁移率族蛋白B1(HMGB1)mRNA的表达水平;免疫印迹法和免疫荧光法检测0、100μmol/L PGZ处理细胞72 h时细胞内HMGB1蛋白表达水平,并采用免疫组化法检测肿瘤增殖标志物Ki67蛋白的表达水平。结果PGZ处理细胞后,培养24 h后,与对照组比较,100、150、200μmol/L PGZ处理组MSTO-211H细胞及150、200μmol/L PGZ处理组NCI-H2452细胞存活率明显下降,差异有统计学意义(P<0.05)。在光镜下可见,与对照组比较,处理组恶性间皮瘤细胞明显减少,形态发生变化。qRT-PCR结果显示,与对照组比较,PGZ处理组PPAR-γmRNA表达水平上升(P<0.05);MSTO-211H细胞中100μmol/L PGZ处理组的HMGB1 mRNA表达水平下降(P<0.05),而NCI-H2452细胞处理组HMGB1 mRNA表达水平上升或无明显变化(P>0.05)。免疫印迹和免疫荧光结果均显示,与对照组比较,MSTO-211H细胞PGZ处理组中HMGB1蛋白表达水平下降(P<0.05),NCI-H2452细胞中HMGB1蛋白表达无明显变化(P>0.05)。免疫组化结果显示,给予PGZ处理后恶性间皮瘤细胞内肿瘤增殖标志物Ki67的表达水平下降。结论PGZ可能通过激活PPAR-γ抑制HMGB1的表达从而抑制恶性间皮瘤细胞的增殖活力。Objective To investigate the effect and mechanism of PPAR-γagonist Pioglitazone(PGZ)on the proliferation of malignant mesothelioma(MM)cells.Methods In December 2019,MM cell lines MSTO-211H and NCI-H2452 were incubated with different final concentrations of PGZ(0,10,50,100,150,and 200μmol/L)for different periods of time(24 h,48 h,and 72 h),and then the cell proliferation level was detected by CCK8 assay.After given various final concentration of PGZ(0,10,50,100,150,200μmol/L)the for 72 hours,the changes of number and morphology of MM cells were observed under an inverted microscope.The expressions of PPAR-γand HMGB1 mRNA were determined by real-time fluorescence quantitative reverse transcription-polymerase chain reaction(qRT-PCR)after treatment of MM cells with PGZ of 0,10,50,100μmol/L for 72 h.The MM cells were treated with PGZ at concentration of 0,100μmol/L for 72 h,and the protein expressions of HMGB1 were examined using Western blotting and immunofluorescence;the protein expressions of Ki67 were assessed by immunohistochemistry.Results The cell viability rate of MM cells was decreased after treated with PGZ(P<0.05).Cell number in PGZ-treated group was significantly less than that in control group and morphology changes were observed under light microscope.QRT-PCR results revealed significantly increased PPAR-γmRNA expression in the PGZ-treated group compared to the control group(P<0.05).There was a significant decrease in the mRNA expression level of HMGB1 in the PGZ-treated group(100μmol/L)as compared to the control group in MSTO-211H(P<0.05);however,the expression level of HMGB1 in NCI-H2452 was an increase or no significant differences(P>0.05).Western blotting and immunofluorescence results showed that the protein expression of HMGB1 was reduced in the PGZ-treated group compared with the control group in MSTO-211H(P<0.05),but the protein expression of that in NCI-H2452 was no significant differences(P>0.05).Immunohistochemistry results showed increased expression of proliferation marker Ki-67.Conc

关 键 词：吡格列酮 恶性间皮瘤 过氧化物酶体增殖物激活受体-Γ 高迁移率族蛋白B1 

分 类 号：R730.26[医药卫生—肿瘤]