健脾清热活血方介导Wnt/β-catenin通路对人结肠SW480细胞的影响  

Treatment with Jian-pi-qing-re-huo-xue detection regulates SW480 cell apoptosis by altering Wnt/ β-catenin expression

作　　者：张保静[1] 张涛[1] 陈远能[1] 王小平[2] 潘怀耿[3] 

机构地区：[1]广西中医药大学附属瑞康医院消化内科,广西壮族自治区南宁市530011 [2]江西中医药高等专科学校,江西省抚州市344700 [3]广州中医药大学,广东省广州市510405

出　　处：《世界华人消化杂志》2013年第9期745-753,共9页World Chinese Journal of Digestology

基　　金：国家自然基金资助项目;No.81001507;No.81260536;广西卫生厅重点基金资助项目;No.2011041~~

摘　　要：目的:观察人结肠癌细胞SW480增殖、凋亡及β-catenin、T细胞因子4(Tcell factor4,TCF-4)、C-myc、CyclinD1表达变化,探讨健脾清热活血方防治结肠癌的可能机制.方法:将SW480细胞分为空白组、治疗组、对照组;采用血清药理学方法,空白组给予胎牛血清;治疗组分别给予等比稀释不同浓度含药血清;对照组给予美沙拉嗪含药血清干预体外培养的结肠癌细胞SW480(24h),应用MTT法检测细胞增殖,流式细胞仪分析细胞凋亡率、细胞周期,免疫荧光染色分析细胞β-catenin蛋白表达,Western blot及荧光定量PCR检测β-catenin、TCF-4、C-myc、CyclinD1蛋白及mRNA表达.结果:与空白组比较,治疗组SW480细胞存活率明显下降(P<0.05),稀释浓度为5%、10%、15%、20%、30%的不同剂量组细胞生长抑制率分别为28.00%、44.58%、65.86%、57.86%、49.89%,但与药物浓度没有剂量依赖性(P>0.05);与对照组比较,治疗组S期细胞增多、G1期细胞减少,且与药物浓度有剂量依赖性(P<0.05);空白组β-catenin表达以核内表达为主,治疗组β-catenin胞质/核异位表达下降(P<0.05),以胞膜表达为主;与空白组比较,治疗组可明显下调β-catenin、TCF-4、C-myc、CyclinD1蛋白及mRNA表达并促进细胞凋亡(P<0.01),以中剂量更为明显.结论:健脾清热活血方可能通过介导Wnt/β-catenin通路,干预SW480细胞G1期,诱导细胞凋亡,发挥防治结肠癌的效应.AIM： To investigate the effect of treatment with Jian-pi-qing-re-huo-xue decoction on SW480 cell proliferation and apoptosis and the expression of β-catenin, TCF-4, c-myc and cyclin D1.METHODS： SW480 cells were divided into a blank group, an intervention group and a control group, which were incubated with fetal calf serum, different concentrations of serum containing Jian-pi-qing-re-huo-xue decoction, or serum containing mesalamine for 24 h, respec-tively. The proliferation and apoptosis of SW480 cells were detected by MMT assay and flow cytometry, respectively. The mRNA and protein expression of β-catenin, TCF-4, c-myc, and cyclin D1 was assayed by real-time PCR and Western blot.RESULTS： Compared to the blank group, the survival of SW480 cells was reduced in the intervention group （P 0.05）. When the dilution concentration of Jian-pi-qing-re-huo-xue decoction was 5%, 10%, 15%, 20% and 30%, the reduced rate of cell growth was 28%, 44.58%, 65.86%, 57.86% and 49.89%, respectively. However, this inhibitory effect was not concentration-dependent （P 0.05）. Intervention with Jian-pi-qing-re-huo-xue decoction significantly increased the percentage of cells at S phase and decreased that of cells at G1 in a concentration-dependent manner compared to the control group （both P 0.05）. β-catenin was more abundantly expressed in the nucleus in the blank group. In the intervention group, the ectopic expression of β-catenin in the cytoplasm and nucleus declined, and membrane expression increased （P 0.05）. Treatment with Jian-pi-qing-re-huo-xue decoction induced cell apoptosis and down-regulated the expression of β-catenin, TCF-4, c-myc, and cyclin D1 compared to the blank group （P 0.01）, and the effect was more significant when middle concentration of serum containing the decoction was used.CONCLUSION： Jian-pi-qing-re-huo-xue decoction exerts a distinct preventive and therapeutic effect on colon cancer possibly by altering the expression of Wnt/ β-catenin, affecting G1 cell cycle progression and

关 键 词：健脾清热活血中药 Β-CATENIN T细胞因子4 C-myc CyclinD1 Wnt Β-CATENIN 凋亡 

分 类 号：R735.34[医药卫生—肿瘤]