目的探讨雌激素是否能激活MAPK信号转导通路,以及对子宫内膜癌细胞增殖能力的影响。方法培养子宫内膜癌Ishikawa细胞,按药物干预分为3组:雌二醇(E_2)组、U0126(MAPK激酶抑制剂)+E_2组、对照组。采用荧光定量PCR技术检测各组MEK1/2、ERK1/2 m RNA表达的变化;Western blot法检测各组p-MEK1/2、p-ERK1/2蛋白的活化程度;流式细胞仪检测各组的细胞周期比例;细胞集落形成实验检测各组细胞的增殖能力;体外穿膜实验比较各组细胞的迁移能力。结果 E_2组MEK1/2、ERK1/2 m RNA表达增加(P=0.025,P=0.002),U0126+E_2组中,U0126能阻断E_2诱导MEK1/2、ERK1/2m RNA的表达上调(P=0.000,P=0.000)。E_2组p-MEK1/2、p-ERK1/2蛋白活化水平升高(P=0.049,P=0.028);U0126+E_2组中,U0126能阻断E_2诱导的p-MEK1/2、p-ERK1/2蛋白活化(P=0.018,P=0.003)。E_2组G1期细胞比例显著低于对照组及U0126+E_2组(P=0.017),集落形成率显著高于对照组及U0126+E_2组(P=0.009),穿过微孔的细胞数显著多于对照组及U0126+E_2组(P=0.000)。结论雌二醇通过激活MAPK通路,促进子宫内膜癌的发展,MEK抑制剂能阻断并抑制这一作用。
Objective:Screening and excavating genes and biological information related to endometrial carcinoma(EC),to provide novel means for clinical diagnosis and treatment.Methods:The gene chip data of EC was obtained from the Gene Expression Omnibus(GEO)database,we integrated gene discrepantly expressive gene analysis,enrichment of genic function and pathway analysis,protein-protein interaction(PPI)network and literature mining to predict genes and pathways of EC.Results:Functional enrichment analysis identified 976differential expression genes were participated in cell cycle regulation,proliferation,biosynthesis,protein phosphorylation and fission biological process,etc.We found eight significantly up-regulated pathways in the microarray datasets,including the MAPK signaling pathway and mTOR pathway,and six down-regulated pathways,including the P53signaling pathway and cell cycle.Gene protein related interaction network analysis was further performed,and we found twelve distinct genes,including the PCNA and CCND1,these genes were located in key position of EC related network,which may be related to the pathogenesis and progression of EC.Furthermore,there was a high correlation between PCNA and CCND1with EC by consulting the related literature and data,however,the functional mechanism was uncleared.Conclusion:The pathogenesis and progression of EC may involve in several different pathways and genes,the genes PCNA,CCND1and MAPK signaling pathway may be valuable.
目的:探讨雌激素能否激活MAPK信号通路,及对子宫内膜癌细胞增殖能力的影响。方法:(1)不同浓度雌二醇作用于HEC-1A细胞后,蛋白印迹法检测p-ERK1/2蛋白活化水平。(2)实验分3组:E2组、U0126+E2组、对照组。荧光定量PCR技术检测细胞中MEK1/2、ERK1/2 m RNA的表达,蛋白印迹法检测p-MEK1/2、p-ERK1/2蛋白活化水平。(3)流式细胞仪,体外穿膜及细胞集落形成实验。结果:(1)不同浓度的E2作用细胞后,蛋白活化水明显高于对照组(P<0.05)。(2)蛋白活化水平E2组均高于对照组(P<0.05);U0126+E2组均低于E2组(P<0.05)。(3)G1期及S期细胞比例;穿过微孔的细胞数;集落形成率U0126+E2组与E2组,E2组与对照组比较,差异均有统计学意义(P<0.05)。结论 :雌二醇通过激活MAPK通路,促进子宫内膜癌的发生、发展。
