莪术提取物榄香烯调控RAF/MEK/ERK信号通路对垂体瘤大鼠细胞生长、分化的影响  

Effects of zedoaria extract elemene on cell growth and differentiation in rats with pituitary tumors by regulating RAF/MEK/ERK signaling pathway

作　　者：张星 熊伟 曹洁 ZHANG Xing;XIONG Wei;CAO Jie(Department of Endocrinology,Wuhan Red Cross Hospital,Wuhan Hubei 430000,China)

机构地区：[1]武汉市红十字会医院内分泌科,湖北武汉430000

出　　处：《临床和实验医学杂志》2020年第21期2245-2248,共4页Journal of Clinical and Experimental Medicine

基　　金：湖北省卫生厅科研项目(编号:JX6B99)。

摘　　要：目的分析莪术提取物榄香烯调控RAF/MEK/ERK信号通路对垂体瘤大鼠细胞生长、分化的影响。方法2019年3月至2019年12月收集购于中国科学院上海细胞生物研究所的大鼠GH3垂体瘤细胞株,采用MTT法、流式细胞仪、蛋白质免疫印迹等方法,分别检测不同质量浓度(0、20、40、60、80μg/m L)的榄香烯处理对大鼠GH3垂体瘤细胞增殖抑制作用、细胞凋亡及其对Raf-1、ERK、Bcl-2、Bax蛋白质表达的影响。结果经20、40、60、80μg/m L质量浓度的榄香烯处理12 h后,GH3细胞的活性[(80.02±8.14)%、(60.01±6.14)%、(41.06±4.14)%、(39.14±3.15)%]依次明显降低,呈剂量依赖性(F=32.045,P<0.05)。60μg/m L质量浓度的榄香烯处理12、24、36、48 h后,GH3细胞的相对活性依次明显降低,呈时间依赖性(F=42.015,P<0.05)。榄香烯呈时间依赖性诱导GH3细胞凋亡,随作用时间增加,凋亡诱导作用增强(F=165.028,P<0.05)。经20、40、60、80μg/m L质量浓度的榄香烯处理GH3细胞2 h后,磷酸化的ERK和Raf-1蛋白表达渐进性降低(F=265.021,P<0.05),但非磷酸化的ERK和Raf-1蛋白表达差异无统计学意义(F=3.021,P>0.05)。榄香烯下调GH3细胞中Bcl-2表达(F=128.27,P<0.05),但Bax表达无显著差异(F=2.328,P>0.05),Bax/Bcl-2值增加(F=106.80,P<0.05)。结论榄香烯可抑制大鼠GH3垂体瘤细胞的体外增殖,抑制RAF/MEK/ERK信号通路,下调下游癌基因Bcl-2表达,增加Bax/Bcl-2值,诱导细胞凋亡,且其作用呈剂量和时间依赖性。Objective To analyze effects of Zedoaria extract elemene on cell growth and differentiation in rats with pituitary tumors by regulating RAF/MEK/ERK signaling pathway.Methods Rat GH3 pituitary tumor cell lines were purchased from Shanghai Institute of Cell Biology,Chinese Academy of Sciences between March and December 2019.Methyl Thiazolyl Tetrazolium(MTT)method,flow cytometry and Western blot were applied to detect effects of different concentrations(0μg/m L,20μg/m L,40μg/m L,60μg/m L,80μg/m L)of elemene on inhibitory effect of GH3 pituitary tumor cell proliferation,cell apoptosis,and expression of Raf-1,ERK,Bcl-2 and Bax proteins.Results After treatment with 20μg/m L,40μg/m L,60μg/m L,80μg/m L mass concentration of elemene for 12 h,the viabilities of GH3 cells[(80.02±8.14)%,(60.01±6.14)%,(41.06±4.14)%and(39.14±3.15)%]were significantly reduced in turn,in a dose-dependent manner(F=32.045,P<0.05).After treatment with 60μg/m L elemene for 12 h,24 h,36 h,and 48 h,the relative activities of GH3 cells were significantly reduced in sequence,which was time-dependent(F=42.015,P<0.05).Elemene induces GH3 cell apoptosis in a time-dependent manner,and the apoptosis-inducing effect increases with the increase of the action time(F=165.028,P<0.05).After treating GH3 cells with 20μg/m L,40μg/m L,60μg/m L,and 80μg/m L mass concentrations of elemene for 2 h,the expression of phosphorylated ERK and Raf-1 protein gradually decreased(F=265.021,P<0.05),but the expressions of non-phosphorylated ERK and Raf-1 protein were not statistically significant(F=3.021,P>0.05).Elemene down-regulated the expression of Bcl-2 in GH3 cells(F=128.27,P<0.05),but there was no significant difference in the expression of Bax(F=2.328,P>0.05),and the value of Bax/Bcl-2 increased(F=106.80,P<0.05).Conclusion Elemene can inhibit proliferation in vitro of GH3 pituitary tumor cells,inhibit RAF/MEK/ERK signaling pathway,down-regulate expression of downstream oncogene Bcl-2,increase Bax:Bcl-2,and induce apoptosis,showing dose-dependent and time-

关 键 词：垂体瘤 榄香烯 RAF-1 ERK Bcl-2 细胞凋亡 

分 类 号：R736.4[医药卫生—肿瘤]