藤龙补中汤促结肠癌LS-174-T细胞衰老及其机制

Senescence-inducing effects of Chinese herbal medicine Tenglong Buzhong Decoction on human colon carcinoma LS-174-T cells and the mechanism

目的:细胞衰老是重要的细胞内在抗癌机制,与肿瘤治疗效应机制密切相关。本研究观察中药复方藤龙补中汤对人结肠癌LS-174-T细胞衰老及相关信号转导的影响。方法:藤龙补中汤处理结肠癌细胞LS-174-T,显微镜观察细胞形态,衰老相关β-半乳糖苷酶(senescence-associatedβ-galactosidase,SA-β-gal)染色检测SA-β-gal活性,流式细胞术检测细胞周期,Western blot法检测p21WAF1/CIP1、p53、p16基因的表达以及RB磷酸化水平。结果:藤龙补中汤作用后,LS-174-T细胞呈现典型大而扁平的衰老形态,SA-β-Gal染色阳性,细胞周期阻滞于G0/G1期。Western blot显示藤龙补中汤可促进LS-174-T细胞p21WAF1/CIP1、p16等基因的表达,对p53、RB等基因的表达无明显影响,可抑制RB磷酸化。结论:藤龙补中汤可促进LS-174-T细胞衰老,并可能与促进p21WAF1/CIP1、p16等基因的表达,以及抑制RB磷酸化相关。

Objective： Cell senescence is an important anti-cancer mechanism and may contribute to cancer therapeutic outcome.The present study observed the effects of Tenglong Buzhong Decoction（TLBZD）,a Chinese herbal formula,on senescence in human colon carcinoma LS-174-T cells.Methods： LS-174-T cells were treated with TLBZD,and morphology change was observed under a microscope.Cell senescence was identified by senescence-associated ？-galactosidase（SA-？-gal） staining,and cell cycle was analyzed by flow cytometry.Expressions of p53,p21WAF1/CIP1,p16 and RB and RB phosphorylation were detected by Western blot.Results： After being treated with TLBZD,LS-174-T cells became enlarged and flattened by morphology;SA-？-gal staining was positive and cell cycle was arrested in G0/G1.In addition,up-regulations of p21WAF1/CIP1 and p16,as well as inhibition of RB phosphorylation were detected in response to TLBZD treatment.Expressions of p53 and RB were unchanged after TLBZD treatment.Conclusion： TLBZD is effective in inducing cell senescence in human colon carcinoma LS-174-T cells,which may relate to up-regulations of p21WAF1/CIP1 and p16 and inhibition of RB phosphorylation.