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目的 :观察藤龙补中汤对人结肠癌RKO细胞衰老及其调控基因表达的影响。方法 :藤龙补中汤处理人结肠癌RKO细胞后,应用衰老相关β-半乳糖苷酶染色法检测细胞衰老情况,PCR芯片检测细胞衰老调控基因转录,蛋白质印迹法检测细胞周期蛋白依赖激酶抑制剂1A(cyclin-dependent kinase inhibitor1A,CDKN1A)/细胞周期蛋白依赖激酶抑制剂2A(cyclin-dependent kinase inhibitor 2A,CDKN2A)-视网膜母细胞瘤蛋白(retinoblastoma protein,RB)-E2F转录因子1(E2F transcription factor 1,E2F1)信号通路中相关蛋白的表达。结果 :藤龙补中汤可促结肠癌RKO细胞衰老(P<0.01),CDKN1A和CDKN2A转录和蛋白表达水平均上调(P值均<0.01),E2F1下游基因细胞周期蛋白A2(cyclin A2,CCNA2)、细胞周期蛋白E1(cyclin E1,CCNE1)和细胞周期蛋白依赖激酶2(cyclin-dependent kinase 2,CDK2)转录和蛋白表达水平均下调(P值均<0.05),RB1蛋白的磷酸化被抑制(P<0.01)。结论 :藤龙补中汤可促进人结肠癌RKO细胞衰老,可能与调控CDKN1A/CDKN2A-RB-E2F1信号通路相关。
Objective: To observe the effect of Fujimura Buzhong decoction on the senescence of human colon cancer RKO cells and its regulation of gene expression. Methods: The decoction of Fujio Buzhong decoction was used to treat human colon cancer RKO cells. Aging-related β-galactosidase staining was used to detect cell senescence. PCR was used to detect the cellular senescence-regulated gene transcription. Western blotting was used to detect the expression of cyclin dependent kinase (CDKN1A) / cyclin-dependent kinase inhibitor 2A (CDKN2A) - retinoblastoma protein (RB) -E2F transcription factor 1 (E2F transcription factor 1, E2F1) signaling pathway. Results: Fuju Bu Buzhong decoction could promote the senescence of RKO cells of colon cancer (P <0.01), the transcription and protein expression of CDKN1A and CDKN2A were up-regulated (P <0.01), and the expression of cyclin A2 and CCNA2 ), Cyclin E1 (CCNE1) and cyclin-dependent kinase 2 (CDK2) were downregulated (all P <0.05). The phosphorylation of RB1 protein was inhibited (P <0.01). Conclusion: Fuju Bu Buzhong decoction can promote the senescence of human colon cancer RKO cells, which may be related to the regulation of CDKN1A / CDKN2A-RB-E2F1 signaling pathway.