木犀草素抑制ERK1/2与p38 MAPK的活化介导鼻咽癌CNE1细胞的生长、移动性和肿瘤干细胞样特性  

Luteolin Mediates of the Growth,Motion and Tumor Stem Cell-Like Properties of Nasopharyngeal Carcinoma CNE1 Cells by Inhibiting the Activation of ERK1/2 and p38 MAPK

作　　者：陈进芬 刘平 陈静 吴志勇 华清泉[2] 王兰[3] CHEN Jinfen;LIU Ping;CHEN Jing;WU Zhiyong;HUA Qingquan;WANG Lan(Yingcheng People's Hospital,Hubei Yingcheng China 432400;Renmin Hospital of Wuhan University,Hubei Wuhan China 430000;Affiliated Hospital of Wuhan University of Science&Technology,Hubei Wuhan China 430000)

机构地区：[1]应城市人民医院,湖北应城432400 [2]武汉大学人民医院,湖北武汉430000 [3]武汉科技大学附属医院,湖北武汉430000

出　　处：《中医学报》2021年第6期1285-1291,共7页Acta Chinese Medicine

基　　金：湖北省自然科学基金项目(ZRMS2017000521)。

摘　　要：目的:探索木犀草素对鼻咽癌CNE1细胞生长、移动性和肿瘤干细胞样特性的影响,及对相关蛋白表达的影响。方法:将CNE1细胞随机分为4组,对照组用含正常血清的DMEM培养基培养,其余3组细胞的血清DMEM培养基中分别加入5、10、20 mg·L^(-1)木犀草素,EdU渗入检测细胞增殖状态;免疫印迹法检测Ki67、Bcl-2/Bax、Survivin、c-Myc、VEGF、Vimentin、Fibronectin的表达及ERK1/2和p38 MAPK的磷酸化水平;Transwell实验观察细胞侵袭情况;肿瘤细胞成球实验检测肿瘤干细胞样特性;RT-PCR检测干细胞标记物CD33、SOX-2、OCT4的mRNA表达水平。结果:木犀草素抑制鼻咽癌CNE1细胞的增殖(P<0.05);与对照组比较,低剂量组Ki67、Bcl-2/Bax和p-ERK1/2蛋白表达水平显著降低(P<0.05),中、高剂量组Ki67、Bcl-2/Bax、Survivin、c-Myc、VEGF、Vimentin、Fibronectin、p-ERK1/2和p-p38 MAPK蛋白表达水平均显著降低(P<0.05);木犀草素处理后侵袭细胞数明显减少,肿瘤干细胞成球直径和数目均显著减少(P<0.05);与对照组比较,低剂量组OCT4 mRNA表达量显著降低(P<0.05),中、高剂量组CD33 mRNA、SOX-2 mRNA、OCT4 mRNA表达量均显著降低(P<0.05)。结论:木犀草素通过抑制ERK1/2和p38 MAPK的磷酸化,抑制鼻咽癌CNE1细胞的生长和运动,降低鼻咽癌干细胞样特性。Objective:To explored the effects of luteolin on the growth,motion,tumor stem cell-like properties of nasopharyngeal carcinoma(NPC)CNE1 cells,and the effect on the expression levels of related proteins.Methods:CNE1 cells were randomly divided into 4 groups,the control group was cultured in DMEM medium with normal serum,and add 5,10,20 mg·L^(-1) luteolin to the serum DMEM medium of the remaining 3 groups respectively.Cell proliferation status was detected by EDU.The expression levels of Ki67,Bcl2/Bax,Survivin,c-Myc,VEGF,Vimentin,Fibronectin and the phosphorylation levels of ERK1/2 and p38 MAPK were detected by Western Blot.Cell invasion was observed by Transwell experiment.Tumor stem cell-like characteristics were detected by tumor cell spheroidization assays.The mRNA expression levels of CD33,SOX-2 and OCT4 were detected by RTPCR.Results:Luteolin can inhibit the proliferation of CNE1 cells(P<0.05).Compared with the control group,the protein expression levels of Ki67,Bcl-2/Bax and p-ERK1/2 in the low-dose group were significantly reduced(P<0.05),and the expression levels of Ki67,Bcl-2/Bax,Survivin,c-Myc,VEGF,Vimentin,Fibronectin,p-ERK1/2 and p-p38 MAPK were significantly decreased in the medium-dose group and high-dose group(P<0.05).The diameter and number of tumor stem cells and invasive cells significantly decreased(P<0.05).Compared with the control group,the levels of OCT4 mRNA in the low-dose group was significantly decreased(P<0.05),and the levels of CD33 mRNA,SOX-2 mRNA and OCT4 mRNA in the medium-dose group and high-dose group were significantly reduced(P<0.05).Conclusion:Luteolin inhibits the growth and motion of CNE1 cells and reduces its stem-like properties by inhibiting the phosphorylation of ERK1/2 and p38 MAPK.

关 键 词：木犀草素 ERK1/2 p38 MAPK 鼻咽癌 生长运动 干细胞样特性 

分 类 号：R285.5[医药卫生—中药学]