半枝莲提取物通过抑制STAT3/SKP2信号通路促进鼻咽癌细胞周期阻滞  

Scutellariae Barbatae Herba Extract Promotes Cycle Arrest of Nasopharyngeal Carcinoma Cells by Inhibiting STAT3/SKP2 Signaling Pathway

作　　者：丁虹[1] 吴紫陆 蔡纪堂[2] 李静波[2] DING Hong;WU Zilu;CAI Jitang;LI Jingbo(Second Clinical Medical College,Henan University of Chinese Medicine,Zhengzhou 450002,China;Henan Province Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南中医药大学第二临床医学院,郑州450002 [2]河南省中医院,郑州450002

出　　处：《中国实验方剂学杂志》2022年第19期81-88,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南中医药大学研究生科研能力创新提升计划项目(2021KYCX025);河南省中医药科学研究专项(2022ZY1057)。

摘　　要：目的:以人鼻咽癌CNE1细胞为研究对象,通过在培养液中添加半枝莲提取物(0.25、0.5、1 g·L^(-1)),来进一步探讨半枝莲提取物对鼻咽癌细胞周期阻滞的影响及其可能的分子作用机制。方法:半枝莲提取物处理CNE1细胞后,采用细胞增殖与活性检测(CCK-8)法检测细胞增殖情况;吉姆萨(Giemsa)染色检测克隆形成率;流式细胞术检测细胞周期分布;并且通过小干扰核糖核酸(siRNA)或者过表达的方式,以逆转录聚合酶链式反应(RT-PCR)和蛋白免疫印迹法(Western blot)检测mRNA的相对表达。结果:与空白组比较,半枝莲提取物组CNE1细胞的增殖和克隆形成率明显降低(P<0.05,P<0.01),呈浓度和时间依赖性。与空白组比较,半枝莲提取物组细胞合成前期(G_(0)/G_(1))升高(P<0.05,P<0.01)。与空白组比较,半枝莲提取物组S期激酶相关蛋白2(SKP2)表达水平显著降低(P<0.01);与半枝莲提取物组比较,过表达SKP2+半枝莲提取物组p21和p27蛋白水平显著降低(P<0.01)。与空白组比较,半枝莲提取物组CNE1细胞中转录激活因子3(STAT3)信号的活化及STAT3磷酸化水平明显降低(P<0.05,P<0.01)。结论:半枝莲提取物能够有效抑制鼻咽癌细胞CNE1的增殖,其机制可能与其作用于STAT3/SKP2途径有关。Objective:To explore the effect of Scutellariae Barbatae Herba extract on on the cycle arrest of nasopharyngeal carcinoma cells and the possible mechanism by adding different concentration of Scutellariae Barbatae Herba extract(0.25,0.5,1 g·L^(-1))in the culture medium,taking CNE1(nasopharyngeal carcinoma cells)as the research object.Method:After the treatment of CNE1 by Scutellariae Barbatae Herba extract,cell counting kit-8(CCK-8)was used to detect cell proliferation,and Giemsa staining was used to detect the clone formation rate.Flow cytometry was used to detect cell cycle distribution,and reverse transcription-polymerase chain reaction(RT-PCR)assay and Western blot assay were used to detect the relative expression of messenger ribonucleic acid(mRNA)by small interfering RNA(siRNA)or overexpression.Result:As compared with the blank group,the proliferation and colony formation rate of CNE1 in the Scutellariae Barbatae Herba extract group significantly decreased(P<0.05,P<0.01)in a dose and time-dependent manner,whereas the percentage of cells in the presynthetic phase(G_(0)/G_(1))increased(P<0.05,P<0.01).The expression level of S-phase kinase associated protein 2(SKP2)in the Scutellariae Barbatae Herba extract group significantly decreased(P<0.01)as compared with the blank group.As compared with the Scutellariae Barbatae Herba extract group,the protein levels of p21 and p27 significantly decreased in the overexpressed SKP2+Scutellariae Barbatae Herba extract group(P<0.01).As compared with the blank group,the signal activation and the phosphorylation level of signal transducer and activator of transcription 3(STAT3)of CNE1 in the S.barbata extract group significantly decreased(P<0.05,P<0.01).Conclusion:Scutellariae Barbatae Herba extract effectively inhibits the proliferation of CNE1,and the mechanism may be related to its action on the STAT3/SKP2 pathway.

关 键 词：半枝莲 鼻咽癌 细胞周期阻滞 转录激活因子3(STAT3) S期激酶相关蛋白2(SKP2) 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学] R2-031R285.5