莪术醇β―环糊精包合物对Bel―7404肝癌细胞体外抑瘤作用和细胞周期影响研究.docVIP

莪术醇β―环糊精包合物对Bel―7404肝癌细胞体外抑瘤作用和细胞周期影响研究 　　[摘要] 目的 探讨莪术醇β-环糊精包合物作用对肝癌细胞的增殖与凋亡及细胞周期的影响及其分子机制。 方法 采用不同浓度莪术醇β-环糊精包合物处理肝癌Bel-7404细胞后，噻唑蓝[3-（4，5）-dimethylthiahiazo （-z-y1）-3，5-di-phenytetrazoliumromide， MTT]比色法测定细胞增殖，流式细胞仪检测细胞周期与凋亡情况。Western blot检测细胞周期调控相关基因的表达水平。 结果 MTT法检测结果显示，与对照组比较，莪术醇β-环糊精包合物各剂量组生长抑制率均明显升高，呈剂量与时间依赖性，差异有统计学意义（P0.05）。流式细胞术检测结果显示，莪术醇β-环糊精包合物处理促进细胞凋亡（P0.05），并使细胞阻滞在G0/G1期（P0.05），伴随p21WAF1及p27KIP1的表达水平升高。 结论 莪术醇β-环糊精包合物能够抑制肝癌细胞Bel-7404的增殖，促进凋亡，并通过上调p21WAF1及p27KIP1基因的表达而诱导G1期阻滞。 　　[关键词] 肝癌；莪术醇；β-环糊精；细胞周期阻滞 　　[中图分类号] R285 [文献标识码] A [文章编号] 1673-9701（2015）16-0011-04 　　[Abstract] Objective To investigate the effects and mechanisms of curcumol-β-cyclodextrin inclusion compound on the proliferation， apoptosis， and cell cycle of human hepatocarcinoma cell line Bel-7404. Methods The effects of different dose of curcumol-β-cyclodextrin inclusion compound on proliferation of human hepatocarcinoma cell line Bel-7404 in vitro was analyzed by MTT assay（P0.05）. The cell cycles and apoptosis were detected by flow cytometer（P0.05）. Expression of cycle regulation-related genes were assessed by Western blot. Results Compared with control groups， MTT results showed that curcumol-β-cyclodextrin inclusion compound significantly inhibited the proliferation of Bel-7404 cells in a dose- and time-dependent manner. The results of flow cytometry showed that curcumol-β-cyclodextrin inclusion compound resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction（P0.05）. Western blotting results showed that curcumol-β-cyclodextrin inclusion compound increased the expression of p21WAF1 and p27KIP1. Conclusion Curcumol-β-cyclodextrin inclusion compound inhibits the growth and induces apoptosis and G0/G1 phase arrest of Bel-7404 in vitro. The mechanisms of G1 arrest may be related to the regulation of the p21WAF1 and p27KIP1. 　　[Key words] Hepatocarcinoma； Curcumol； β-cyclodextrin； Cell cycle arrest 　　原发性肝癌是我国最常见的恶性肿瘤之一，一旦确诊多数患者已为晚期，丧失手术治疗的最佳时机[1]。尽管肝癌的综合