葛根素对子宫内膜异位症细胞恶性行为调控机制研究

Regulatory mechanism of malignant behavior of endometriosis mediated by puerarin

目的:观察葛根素对子宫内膜异位症(endometriosis,EM)血管形成的影响以及葛根素对EM肿瘤相关基因的调控作用。方法:利用子宫内膜细胞原代培养技术、鸡胚绒毛尿囊膜(chicken chorioallantoic membrane,CAM)模型、基因芯片等方法,了解EM肿瘤相关基因表达的情况,观察葛根素对EM血管生成、肿瘤相关基因的调控作用。结果:葛根素可明显抑制种植内膜组织囊泡及周边血管形成,其血管阳性面积显著小于模型组(P<0.05)。基因芯片结果显示,增殖期EM在位内膜组织较之正常在位内膜组织、EM异位内膜组织较之EM在位内膜组织,在癌基因,抑癌基因,细胞黏附、侵袭、转移、血管生成相关基因及细胞凋亡相关基因表达方面有明显不同。100μmol/L葛根素可明显上调EM在位基质细胞凋亡基因BAD、BAX、CASP8、CASP9和TNFRSF6表达,并增强细胞周期依赖性蛋白激酶抑制因子CDKN1B、CDKN2A及IFNA1、IFNB1基因表达;减少癌基因FOS、CHEK2、SRC和黏附侵袭基因ITGB5、MMP9及生长、转录因子基因PDGFA、NFKBIA表达。结论:葛根素能够抑制异位灶新生血管的形成,减少异位灶的血液供应;通过调整EM癌基因、抑癌基因、细胞周期相关因子基因表达,并通过影响凋亡相关基因的表达,促进细胞凋亡,从而改变EM内膜细胞类似肿瘤细胞的恶性行为。

Objective： To observe the inhibitory effects of puerarin on angiopoiesis of endometriotic tissue, and to explore the regulatory effects of puerarin on tumor-related gene expression of endometriosis. Metheds：The regulatory effects of puerarin on endometriotic angiopoiesis and tumor-related gene expression were observed by using a chicken chorioallantoic membrane model and gene array method. Results： Chicken chorioallantoic membrane experiment indicated that puerarin obviously inhibited endometriotic vasiculation and angiopoiesis. The area of blood vessels was significantly reduced as compared with the untreated group （P 〈0. 05）. The expressions of oncogenes and genes related to adhesion, invasion, and apoptosis, including ERBB2, CD44, ITGA6, NCAM1, MMP1, FLT1, AKT1, BCL2L ETS2, FOS, S100A4, TEK, TERT, NFKBIA, CDH1, and BIRC5 genes, were obviously higher, while the expressions of the anti-oncogenes, anti-apoptosis genes and anti-invasion genes, including KAI1, KISSI, SERPINBS, TNFRSF25, TNFRSF1A, TNFRSF6 and SERPINB2, were significantly lower in eutopic endometrial tissue from patients with endometriosis than those from endometriosis-free women. The expressions of oncogenes （ERBB2, ETS2, FQS）, apoptosis gene （BCL2L1）, cyclin-dependent kinases （CDK4, CDC25A）, and growth factor and receptors （HGF, FGFR2, TGFBR） were significantly enhanced, while the expressions of theanti-oncogenes （KAI1, SERPINB5）, apoptosis genes （BAD and TNF） and cyclin-dependent kinase inhibiting factor （CDKN2A） were obviously reduced in ectopic tissue as compared with those in eutopic tissue from patients with endometriosis. Puerarin significantly enhanced the gene expressions in endometriotic stromal cells, including BAD, BAX, CASPS, CASPg, TNFRSF6, CDKNIB, CDKN2A, IFNA1 and IFNBI, and reduced the gene expressions of FOS, CHEK2, SRC, ITGBS, MMP9, PDGFA and NFKBIA. Conclusions： The tumor-related gene expression has significant differences in eutopic endometrial tissue between patients with endometriosis and endometriosis-free women, and between ectopic and eutopic tissues from patients with endometriosis. Puerarin can reduce angiopoiesis, regulate tumor-related gene expression and facilitate apoptosis in endometriotic tissue.