丹皮酚对人大肠癌细胞凋亡和细胞周期以及患者预后的影响

Effect of paeonol on apoptosis and cell cycle of human colorectal cancer cells as well as prognosis of patients

目的:探讨中药丹皮酚(paeonol,Pae)对人大肠癌细胞株HT-29凋亡和细胞周期的影响,及其与化疗药物的协同作用,为寻求新的大肠癌治疗策略提供理论依据。方法:实验于2004-05/09在武汉大学人民医院消化病研究所完成;应用MTT法、流式细胞仪技术检测不同浓度的丹皮酚对HT-29细胞的增殖抑制、凋亡诱导作用及对细胞周期分布的影响,同时观察低浓度的丹皮酚与化疗药物的协同作用。结果:丹皮酚(浓度7.81～250mg/L)可抑制HT-29细胞增殖,并且与药物浓度及作用时间呈正相依赖关系,最高抑制率为91.849%(P<0.01)。丹皮酚在15.63,62.5,250mg/L3种浓度下作用48h均可诱导HT-29细胞凋亡,流式细胞仪技术检测其凋亡率分别为7.6%,16.2%,和34.5%,有明显的剂量效应关系。同时丹皮酚使细胞周期分布发生明显变化,表现为S期细胞比例上升,G0/G1期和G2/M期细胞比例下降。低浓度的丹皮酚与化疗药物协同可产生较强的抑制细胞增殖作用,其中与5氟尿嘧啶的协同作用最为显著。结论:丹皮酚能抑制HT-29细胞增殖并诱导其发生凋亡,其机制可能与影响该细胞株的细胞周期有关。丹皮酚与细胞周期特异性化疗药联合使用具有协同抗肿瘤作用。

AIM:To investigate the effect of paeonol(Pae) on apoptosis and cell cycle of human colorectal cancer cell line HT 29 and its synergetic effect with chemotherapy agents so as to provide an theoretical basis for new therapy for colorectal cancer. METHODS:The study was conducted in the Institute of Gastroenterology,Renmin Hospital of Wuhan University from May to September 2004.The inhibitory effect of Pae on HT 29 cell proliferation was detected by MTT assay.The induction of apoptosis and the influence on the distribution of HT 29 cell cycle were measured by flow cytometry.The synergetic effect of Pae at low concentration with chemotherapy agents was observed simultaneously. RESULTS:Pae at the concentration of 7.81 to 250 mg/L could inhibit the proliferation of HT 29 cells,which showed obvious concentration effect and time effect relationship,and the highest inhibition rate was 91.849% (P< 0.01).Pae at the concentration of 15.63,62.5 and 250 mg/L could induce apoptosis of HT 29 cells after 48 hours.By flow cytometry(FCM) methods,the apoptosis rates of HT 29 cells were 7.6% ,16.2% and 34.5% respectively,which showed obvious concentration effect relationship.The distribution of cell cycle was altered simultaneously.S phase of the cells was increased,while G0/G1 and G2/M phase of the cells were decreased after treated by Pae.Pae at low concentration with several chemotherapy agents,especially with 5 FU,had synergetic effect in inhibiting the proliferation of HT 29 cells. CONCLUSION:Pae may inhibit proliferation of HT 29 cells and induce its apoptosis.The possible mechanism is the influence on distribution of HT 29 cell cycle.Pae has synergetic antitumor effect with cell cycle specific chemotherapy agents.