肉桂醛调控CD44s/STAT3信号通路抑制胰腺癌细胞增殖和肿瘤细胞干性  

Cinnamaldehyde inhibits pancreatic cancer cell proliferation and stemness via modulation of CD44s/STAT3 signaling pathway

作　　者：朱素丽 陈运昊 姚尖平[3] 周兴旺[1] ZHU Su-li;CHEN Yun-hao;YAO Jian-ping;ZHOU Xing-wang(Dept of Biochemistry and Molecular Biology,Zhongshan Medical College,the First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China;Dept of Clinical Laboratory,the First Affiliated Hospital,the First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China;Dept of Cardiac Surgery,the First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]中山大学中山医学院生化与分子生物学教研室,广东广州510080 [2]中山大学附属第一医院检验科,广东广州510080 [3]中山大学附属第一医院心外科,广东广州510080

出　　处：《中国药理学通报》2022年第5期676-683,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82174022);广东省基础与应用基础研究基金重点项目(No 2019B1515120051);广州市科技计划项目(No 201803010027)。

摘　　要：目的探究肉桂醛(cinnamaldehyde,CA)对胰腺癌PANC-1细胞的增殖、肿瘤细胞干性的影响及其可能的作用机制。方法采用CCK8法检测不同浓度(0、5、15、20、30、50、70、100、150μmol·L^(-1))和不同时间段(24、48、72 h)CA处理对PANC-1细胞存活率的影响;CFSE染色实验检测CA对PANC-1细胞的增殖抑制作用;克隆形成实验检测CA对单个细胞增殖的影响;利用悬浮干细胞成球实验检测CA对成球能力的影响;qRT-PCR和Western blot检测干性相关Nanog、Sox-2及Oct-4基因的表达;流式细胞术分析CA处理前后的CD44^(+)CD24^(+)和ALDH^(+)干细胞亚群比例;Western blot分析CD44s、p-STAT3及STAT3蛋白表达。结果CA可呈浓度和时间依赖性的降低PANC-1细胞的存活率,并抑制肿瘤细胞增殖能力,明显抑制其克隆形成能力;干细胞成球实验结果显示,CA可使细胞成球体积减小,数量变少;CA处理后,细胞的3种干性基因的mRNA和蛋白表达水平均明显降低;CA处理还可减少癌细胞中CD44^(+)CD24^(+)和ALDH^(+)干细胞亚群比例;Western blot结果表明CA可下调CD44s和p-STAT3的表达水平,而对STAT3蛋白表达无影响,加入STAT3激动剂Colivelin TFA可部分回复CA对癌细胞增殖和肿瘤细胞干性的抑制作用。结论CA可显著抑制胰腺癌PANC-1细胞的增殖和肿瘤细胞干性,其机制可能与调控CD44s/STAT3信号通路相关。Aim To explore the effects of cinnamaldehyde on the proliferation and stemness of pancreatic cancer PANC-1 cells,and its possible mechanism of action.Methods CCK8 assay was used to detect the effect of cinnamaldehyde treatment on cell viability at different concentrations(0,5,15,20,30,50,70,100,150μmol·L^(-1))and different intervention time(24,48,72 h).CFSE proliferation assay was used to detect the inhibitory effects of cinnamaldehyde on PANC-1 cells.Colony formation assay was employed to determine the colony-forming ability of PANC-1 cells after cinnamaldehyde treatment.The sphere formation assay was employed to detect the effects of cinnamaldehyde on sphere-forming ability in PANC-1 cells.Western blot analysis and qRT-PCR analysis were applied to determine the expression levels of Nanog,Sox-2 and Oct-4.Flow cytometry was used to detect the percentage of CD44^(+)CD24^(+) cells and ALDH^(+) cells in cinnamaldehyde treated and untreated PANC-1 cells.Western blot analysis was used to detect the effects of cinnamaldehyde on the expression of CD44s,p-STAT3 and STAT3 in PANC-1 cells.Results Cinnamaldehyde suppressed cell viability in a dose-and time-dependent manner,and inhibited tumor-cell proliferation and colony forming ability significantly in a dose-dependent manner in PANC-1 cells.Sphere-forming assay showed that cinnamaldehyde could significantly inhibit sphere-forming ability in suspension culture of PANC-1 cells.The mRNA and protein expression levels of three stemness-related genes were down-regulated after cinnamaldehyde treatment.In addition,cinnamaldehyde treatment significantly decreased the proportion of CD44^(+)CD24^(+) cells and ALDH^(+) cells.Western blotting showed that cinnamaldehyde inhibited the expression of CD44s and p-STAT3,while it had no effect on the expression of STAT3.With the addition of STAT3 activator(Colivelin TFA),the inhibition of cinnamaldehyde on proliferation and tumor-cell stemness in PANC-1 cells was partially rescued.Conclusions Cinnamaldehyde significantly inhibits the prol

关 键 词：肉桂醛 胰腺癌 PANC-1细胞 增殖 肿瘤细胞干性 CD44s/STAT3信号通路 

分 类 号：R284.1[医药卫生—中药学] R329.28[医药卫生—中医学]