甲基莲心碱抑制腺样囊性癌生长的机制研究  

Study on the mechanism of inhibiting the growth of adenoid cystic carcinoma by neferine

作　　者：李立恒[1] 周军 张敏[4] 王芹[1] 安峰[1] 汪佳[1] 张凡[2] LI Li-heng;ZHOU Jun;ZHANG Min;WANG Qin;AN Feng;WANG Jia;ZHANG Fan(Department of Stomatology,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei Province,China;Department of Pathology,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei Province,China;Department of Stomatology,The First Hospital of Zhangjiakou,Zhangjiakou 075041,Hebei Province,China;Clinical College,Zhangjiakou University,Zhangjiakou 075000,Hebei Province,China)

机构地区：[1]河北北方学院附属第一医院口腔科,河北张家口075000 [2]河北北方学院附属第一医院病理科,河北张家口075000 [3]张家口市第一医院口腔科,河北张家口075041 [4]张家口学院临床学院,河北张家口075000

出　　处：《中国临床药理学杂志》2022年第16期1910-1914,共5页The Chinese Journal of Clinical Pharmacology

基　　金：张家口市科学技术研究与发展计划基金资助项目(1821056D)。

摘　　要：目的研究甲基莲心碱对腺样囊性癌生长的作用及其作用机制。方法涎腺腺样囊性癌细胞SACC-83随机分为对照组、甲基莲心碱低剂量组、甲基莲心碱中剂量组、甲基莲心碱高剂量组。甲基莲心碱低、中、高剂量组分别用10,20,40μmol·L^(-1)甲基莲心碱处理细胞,对照组细胞用等量的培养基处理。以噻唑蓝(MTT)法检测各组细胞增殖情况;以流式细胞术检测各组细胞凋亡情况;以Transwell检测细胞迁移;以蛋白质印迹(Western Blot)法检测高温需求因子A2(Omi/HtrA2)、X染色体连锁的凋亡抑制蛋白(XIAP)表达水平以及增殖、凋亡、迁移相关蛋白表达水平。结果对照组和甲基莲心碱低、中、高剂量组细胞凋亡率分别为(3.31±0.68)%,(13.43±1.76)%,(24.71±2.43)%和(42.37±2.95)%,迁移细胞数分别为134.18±8.28,94.86±6.90,68.81±6.43和50.24±5.27。对照组细胞中Omi/HtrA2蛋白、XIAP蛋白、细胞周期蛋白D1(CyclinD1)、切割型胱天蛋白酶-3(cleaved caspase-3)蛋白和基质金属蛋白酶-9(MMP-9)蛋白的表达水平分别为0.91±0.08,0.73±0.03,0.78±0.09,0.22±0.04和0.93±0.07,不同剂量甲基莲心碱作用后Omi/HtrA2蛋白、XIAP蛋白、CyclinD1蛋白和MMP-9蛋白的表达水平明显降低,cleaved caspase-3蛋白表达水平明显升高,差异均有统计学意义(均P<0.05),且甲基莲心碱对SACC-83细胞的作用具有剂量依赖性。结论甲基莲心碱可以通过调控Omi/HtrA2和XIAP蛋白对腺样囊性癌细胞的生长起到抑制作用。Objective To investigate the effect and mechanism of neferine on the growth of adenoid cystic carcinoma.Methods SACC-83 of salivary adenoid cystic carcinoma cells were randomly divided into the control group,neferine-L group,neferine-M group and neferine-L group.The cells in the neferine-L,-M,-H groups were treated with neferine at the final concentration of 10,20 and 40μmol·L^(-1),respectively.The cells in control group were treated with the same amount of medium.MTT was used to test cell proliferation in each group.The apoptotic cells were detected by flow cytometry.Transwell detect cell migration.The expression levels of high temperature requirement factor A2(Omi/HtrA2)and X-linked apoptosis suppressor protein(XIAP)proteins as well as the expression levels of proliferation,apoptosis and migration-related proteins were detected by Western Blot.Results The cell apoptosis rates in control group,neferine-L group,neferine-M group and neferine-H group were(3.31±0.68)%,(13.43±1.76)%,(24.71±2.43)% and(42.37±2.95)%,respectively and the number of migrated cells in each group was 134.18±8.28,94.86±6.90,68.81±6.43 and 50.24±5.27,respectively.The expression levels of Omi/HtrA 2 protein,XIAP protein,CyclinD1,cleaved caspase-3 protein and matrix metalloproteinase-9(MMP-9)protein in the cells of the control group were0.91±0.08,0.73±0.03,0.78±0.09,0.22±0.04 and 0.93±0.07,respectively.After treated with different doses of neferine,the expression levels of Omi/HtrA 2 protein,XIAP protein,CyclinD1 protein and MMP-9 protein were significantly decreased,and the expression level of cleaved caspase-3 protein was significantly increased.The differences were statistically significant(all P<0.05),and the effect of neferine on SACC-83 cells was dose-dependent.Conclusion Neferine can inhibit the growth of adenoid cystic carcinoma cells by regulating the Omi/HtrA 2 and XIAP proteins.

关 键 词：甲基莲心碱 高温需求因子A2(Omi/HtrA2) X染色体连锁的凋亡抑制蛋白(XIAP) 涎腺腺样囊性癌 增殖 凋亡 

分 类 号：R28[医药卫生—中药学]