组胺通过Hippo-YAP信号通路影响鼻咽癌细胞增殖与侵袭的机制研究  

Effect of histamine on proliferation and invasion of nasopharyngeal carcinoma cells via Hippo-YAP signaling pathway

作　　者：冼德生 曾银萍 周小柳[1] 李智群[1] 王丽萍 羊俊维 XIAN Desheng;ZENG Yinping;ZHOU Xiaoliu;LI Zhiqun;WANG Liping;YANG Junwei(Department of Otorhino-laryngology,First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院耳鼻咽喉头颈外科,海口570102

出　　处：《中国免疫学杂志》2022年第17期2096-2100,2106,共6页Chinese Journal of Immunology

基　　金：海南省医药卫生科研项目(1901031017A2001)。

摘　　要：目的:探讨组胺对鼻咽癌细胞CNE1增殖、侵袭及线粒体膜电位的影响,及其对Hippo-YAP信号的调控作用。方法:细胞实验分为正常组、组胺低、中、高剂量组。正常组细胞不添加任何药物常规培养;组胺低、中、高剂量组细胞依次添加100、200、400μmol/L的组胺处理后进行培养。CCK-8法检测各组CNE1细胞的增殖率,Transwell小室检测各组细胞的侵袭能力,JC-1染色检测各组CNE1细胞的线粒体膜电位,皮下移植瘤裸鼠模型检测各组CNE1细胞的体内生长能力,特异性PCR检测各组CNE1细胞中大肿瘤抑制基因1(LATS1)的甲基化水平,Western blot检测各组CNE1细胞YAP1、p-YAP1、LATS1表达水平。结果:与正常组相比,组胺低、中、高剂量组细胞的增殖率、侵袭细胞数、红色荧光与绿色荧光比值(线粒体膜电位)、肿瘤瘤体质量、LATS1的甲基化比例、YAP1、p-YAP1表达明显降低(均P<0.05),LATS1表达明显升高(均P<0.05),且具有明显的剂量依赖性(均P<0.05)。结论:组胺能抑制鼻咽癌CNE1细胞的体外增殖与侵袭能力,降低其线粒体膜电位,使肿瘤细胞体内生长明显受限,这可能是通过调控Hippo-YAP信号实现的。Objective:To investigate effect of histamine on proliferation,invasion and mitochondrial membrane potential of nasopharyngeal carcinoma cell CNE1,and its regulation of Hippo-YAP signaling pathway.Methods:Cell experiment was divided into normal group,histamine low-,medium-and high-dose group. Cells in normal group were without any drugs with conventional culture.Cells in histamine low-,medium-and high-dose groups were treated with 100,200 and 400 μmol/L histamine and then cultured. CCK-8was used to detect cell proliferation rate of CNE1 cell in each group. Transwell chamber was used to detect invasion ability of CNE1cell in each group. JC-1 staining was used to detect mitochondrial membrane potential of CNE1 cell in each group. Subcutaneous xenograft nude mice model was used to detect growth ability of CNE1 cell in each group in vivo. Specific PCR was used to detect methylation level of large tumor suppressor gene 1(LATS1)in CNE1 cells in each group,and Western blot was used to detect expression levels of YAP1,p-YAP1 and LATS1 in CNE1 cells in each group.Results:Compared with normal group,proliferation rate,number of invaded cells,ratio of red fluorescence to green fluorescence(mitochondrial membrane potential),mass of tumor,ratio of LATS1methylation,expressions of YAP1 and p-YAP1 were significantly decreased in histamine low-,medium-and high-dose groups(all P<0.05),and expression of LATS1 was significantly increased(all P<0.05),all with a significant dose-dependent(all P<0.05).Conclusion:Histamine can inhibit the in vitro proliferation and invasion of nasopharyngeal carcinoma CNE1 cells,reduce their mitochondrial membrane potential,and significantly restrict the growth of tumor cells in vivo,which may be achieved by regulating HippoYAP signaling.

关 键 词：组胺 鼻咽癌 增殖与侵袭 线粒体膜电位 Hippo-YAP信号通路 

分 类 号：R739.6[医药卫生—肿瘤]