毛兰素对口腔鳞状细胞癌细胞增殖和凋亡的影响及机制  

Effects and mechanisms of Erianin on proliferation and apoptosis in oral squamous cell carcinoma cells

作　　者：王瑞瑞 谢李[3] 王基栋 WANG Ruiru;XIE L;WANG Jidong(Department of Oral and Maxillofacial Surgery,Changde Hospital,Xiangya School of Medicine,Central South University(The First People's Hospital of Changde City),Changde 415000,China;Department of Radiology,the 3rd Xiangya Hospital of Central South University,Changsha 410013,China;Department of Head and Neck Surgery,Hunan Cancer Hospital/the Affiliated Cancer Hospital of Xiangya School of Medicine,Changsha 410013,China)

机构地区：[1]中南大学湘雅医学院附属常德医院(常德市第一人民医院)口腔颌面外科,湖南常德415000 [2]中南大学湘雅三医院放射科,湖南长沙410013 [3]湖南省肿瘤医院/湘雅医学院附属肿瘤医院头颈外科,湖南长沙410013

出　　处：《口腔疾病防治》2025年第3期186-194,共9页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：湖南省自然科学基金项目(2024JJ9261)。

摘　　要：目的探讨毛兰素(Erianin)对人口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞增殖和凋亡的影响,为OSCC的临床治疗提供研究基础。方法使用0、2.5、5、10μmol/L浓度的毛兰素处理OSCC细胞CAL27和SCC9;采用CCK-8和软琼脂集落形成实验检测毛兰素对OSCC细胞增殖的抑制作用。通过蛋白免疫印迹法(Western blot)分析毛兰素在OSCC细胞中对抗凋亡相关蛋白B细胞淋巴瘤-超大(B-cell lymphoma-extra large,Bcl-xL)、B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、髓样细胞白血病-1(myeloid cell leukemia-1,Mcl-1)及凋亡蛋白裂解型半胱天冬酶3(cleaved-Caspase 3,c-Caspase 3)表达的影响。通过caspase 3活性检测试剂盒,进一步分析毛兰素在OSCC细胞中的促凋亡效应。通过质粒转染构建过表达Mcl-1的CAL27细胞,并通过Western blot和caspase 3活性测定分析Mcl-1对毛兰素作用于CAL27细胞的影响。获得湖南省肿瘤医院动物实验伦理委员会审批,构建CAL27裸鼠移植瘤模型并随机分为两组(n=5),给药组腹腔注射毛兰素(25 mg/kg),对照组注射磷酸盐缓冲液(phosphate buffered saline,PBS)。利用免疫组化方法检测肿瘤组织中Ki67和Mcl-1的表达水平。结果毛兰素呈剂量依赖性地抑制CAL27和SCC9细胞的增殖,并下调了Mcl-1的蛋白表达,而对Bcl-2和Bcl-xL的影响较小。此外,毛兰素诱导OSCC细胞发生凋亡,表现为c-Caspase 3蛋白表达水平增加,caspase 3的活性增强(P<0.001)。过表达Mcl-1则可以抑制毛兰素诱导的c-Caspase 3蛋白水平的增加及caspase 3活性水平。体内实验结果与体外一致,毛兰素处理后,CAL27细胞在裸鼠体内的生长受到抑制(P<0.001),肿瘤组织中增殖相关分子Ki67及抗凋亡分子Mcl-1的表达水平下调(P<0.001)。结论毛兰素具有较好的抗肿瘤效果,能够有效抑制OSCC细胞的增殖并促使其凋亡,其机制可能涉及下调促存活蛋白Mcl-1的表达水平。Objective To investigate the effects of Erianin on cell proliferation and apoptosis in human oral squa-mous cell carcinoma(OSCC)cells,providing a research foundation for the clinical treatment of OSCC.Methods Eri-anin was applied to OSCC cells(CAL27 and SCC9)at concentrations of 0,2.5,5,and 10μmol/L.The inhibitory effect of Erianin on OSCC cell proliferation was evaluated using CCK-8 and soft agar colony formation assays.Western blot-ting(WB)was employed to analyze the expression levels of anti-apoptotic proteins B-cell lymphoma-extra large(Bcl-xL),B-cell lymphoma-2(Bcl-2),myeloid cell leukemia-1(Mcl-1),and apoptotic protein cleaved-Caspase 3(c-Caspase 3)in OSCC cells.Caspase 3 activity was further assessed using a caspase 3 activity detection kit to examine the pro-apoptotic effect of Erianin in OSCC cells.Mcl-1 overexpression was induced in CAL27 cells via plasmid transfection,and the in-fluence of Mcl-1 on the effects of Erianin in CAL27 cells was analyzed by WB and caspase 3 activity measurement.All animal experiments were approved by the Ethics Committee of Hunan Cancer Hospital.A CAL27 xenograft mouse mod-el was established and randomly divided into two groups(n=5):the treatment group received intraperitoneal injection of Erianin(25 mg/kg),while the control group was injected with phosphate-buffered saline(PBS)as the vehicle.Immu-nohistochemistry(IHC)was used to detect the expression levels of Ki67 and Mcl-1 in the tumor tissues.Results Eri-anin inhibited the proliferation of CAL27 and SCC9 cells in a dose-dependent manner and downregulated the protein ex-pression of Mcl-1,with minimal effects on Bcl-2 and Bcl-xL.Furthermore,Erianin induced apoptosis in OSCC cells,as evidenced by increased expression of c-Caspase 3 and enhanced caspase 3 activity(P<0.001).Overexpression of Mcl-1 inhibited the Erianin-induced increase in c-Caspase 3 protein levels and caspase 3 activity.In vivo results were consis-tent with the in vitro findings.After Erianin treatment,CAL27 cell growth in nude mice was suppressed(P<0.001),

关 键 词：口腔鳞状细胞癌 毛兰素 天然化合物 促存活蛋白 髓样细胞白血病-1 细胞凋亡 细胞增殖 抗肿瘤机制 

分 类 号：R78[医药卫生—口腔医学]