摘要
目的探究氧化苦参碱通过调控SIRT1-TSC2抑制人子宫内膜癌HEC-1B细胞生长及Wnt信号通路。方法采取MTT法对不同浓度50、100、200、400μmol/L下氧化苦参碱对HEC-1B细胞抑制率;氧化苦参碱+pcDNA3.1组(转染pcDNA3.1)、氧化苦参碱+pcDNA3.1-SIRT1-TSC2组(转染pcDNA3.1-SIRT1)均用脂质体法转染,加入氧化苦参碱(200μmol/L)处理;应用实时荧光定量PCR(qRT-PCR)法、Western blotting法对细胞内信息调节因子1(SIRT1)、TSC2、β-catenin、c-Myc的mRNA水平与蛋白表达分别进行检测和评估,予以流式细胞术对细胞凋亡率进行检测。结果相比对照组,予以不同浓度50、100、200、400μmol/L下氧化苦参碱各组的细胞增殖抑制率与凋亡率均呈增高趋势(P<0.05),氧化苦参碱(200μmol/L)组细胞内SIRT1、TSC2的mRNA和蛋白水平均呈显著降低趋势(P<0.05);过表达SIRT1-TSC2可逆转氧化苦参碱对HEC-1B细胞增殖、Wnt信号通路失活效果。结论氧化苦参碱可调控SIRT1-TSC2抑制子宫内膜癌细胞增殖,促进凋亡,其促凋亡机制可能与失活Wnt信号通路有关,为子宫内膜癌的早期诊治提供新的策略。
Objective To investigate the inhibitory effect of oxymatrine on the growth and Wnt signaling pathway of human endometrial carcinoma HEC-1 B cells by regulating SIRT1-TSC2.Methods MTT method was used to determine the the inhibitory rate of oxymatrine on HEC-1 B cells at concentrations of 50,100,200 and 400μmol/L.Oxymatrine+pcDNA3.1 group(transfected pcDNA3.1)and oxymatrine+pcdna3.1-sirt1-tsc2 group(transfected pcdna3.1-sirt1)were transfected by liposome method,and oxymatrine(200μmol/L)was added.The mRNA levels and protein expressions of intracellular information regulatory factor 1(SIRT1),TSC2,β-catenin and c-Myc were detected and evaluated respectively,and the apoptosis rate was detected by flow cytometry.Results Compared with those of the control group,the cell proliferation inhibition rates and apoptosis rates of oxymatrine with concentrations of 50,100,200 and 400μmol/L were increased(P<0.05),and the mRNA and protein levels of SIRT1 and TSC2 of oxymatrine(200μmol/L)group decreased significantly(P<0.05).Over-expression of SIRT1-TSC2 could reverse the effects of oxymatrine on the proliferation and inactivation of Wnt signaling pathway of HEC-1 B cells.Conclusion Oxymatrine can regulate SIRT1-TSC2,inhibit the proliferation of endometrial cancer cells and promote apoptosis.Its mechanism of promoting apoptosis may be related to the inactivation of Wnt signal pathway,which provides a new strategy for the early diagnosis and treatment of this disease.
作者
鲁强
李霞
管东方
周晓
LU Qiang;LI Xia;GUAN Dongfang;ZHOU Xiao(Qingdao Hospital of Traditional Chinese Medicine(Qingdao Hiser Hospital),Qingdao 266033,Shandong,China)
出处
《中华中医药学刊》
CAS
北大核心
2022年第12期240-243,共4页
Chinese Archives of Traditional Chinese Medicine
基金
山东省医药卫生科技发展计划(2016WS0303)
青岛市医药科研指导计划(2015-WJZD033)。
