从全基因表达谱角度探讨理冲汤抑制人子宫肌瘤细胞的作用机制

摘要

目的：从全基因表达谱角度探讨理冲汤抑制人子宫肌瘤细胞的作用机制。方法采用胶原酶消化法培养子宫肌瘤细胞,并给予理冲汤大鼠含药血清进行干预,提取肌瘤细胞总RNA,运用Affymetrix GeneChip PrimeViewTM全基因芯片检测基因表达谱的变化,并进行差异表达基因、基因本体(gene ontology,GO)、信号通路等方面的分析,最后采用实时定量PCR(Realtime PCR, RT-PCR)对差异基因的表达进行验证。结果结果显示,理冲汤组与空白对照组比较后差异表达基因数量为1154个(P<0．05,上调基因Fold change>1．5,下调基因Fold change<0．5),其中上调基因630个,下调基因524个；富集的GO功能共有119条,主要涉及到细胞外基质的改变、细胞凋亡的诱导、细胞黏附等多方面的功能；差异基因富集的信号通路有73条,同时,RT-PCR验证结果也与基因芯片结果趋势一致。结论理冲汤对人子宫肌瘤的治疗作用机制是通过对多基因、多功能、多信号通路的调控而实现的。%Objective To investigate the molecular mechanism of Lichong Decoction ( LCD ) inhibiting human uterine leiomyoma cells through the analysis of differential gene expression profiles of uterine leiomyoma cells. Methods Using collagenase digestion cultured uterine leiomyoma cells,and trea-ted by rat serum containing of LCD for 48 hours. Then the pathological changes of uterine leiomyoma cells were observed, and total RNA of uterine leiomyoma cells were extracted. The gene expression profile was detected by whole genome chip testing, then differentially expressed genes (DEGs), gene ontology(GO) and signal pathway were analyzed. The mRNA expressions were verified by real-time PCR (RT-PCR). Results Compared with the control group, the growth of LCD group cells significantly inhibited by micro-scope. The DEGs between the LCD group and the control group ( P <0. 05, Up Regulated Genes Fold change>1. 5, Down Regulated Genes Fold change<0. 5) were 1154, included 630 up-regulated and 524 down-regulated genes. The GO enrichment analysis (P<0. 05) showed 119 functions, included extracellu-lar matrix, induction of apoptosis, negative regulation of angiogenesis, cell adhesion, and so on. The sig-nal pathway analysis showed 73 enriched pathways ( P<0. 05 ) , The verification of mRNA expression by RT-PCR was consistent with that of microarray. Conclusion LCD treats uterine leiomyoma through the regulation of multiple genes and multiple signal pathways.