莪术醇β-环糊精包合物对食管癌TE-1细胞增殖、凋亡的影响及其机制研究

Effects and Mechanisms of Curcumol β-cyclodextrin Compound on the Proliferation and Apoptosis of Esophageal Carcinoma Cell Line TE-1

目的探讨莪术醇β-环糊精包合物对食管癌TE-1细胞增殖、凋亡的影响及其作用机制。方法采用饱和溶液法制备莪术醇与β-环糊精的包合物,红外光谱验证包合物的形成。不同浓度(25、50、100mg/L)莪术醇β-环糊精包合物处理食管癌TE-1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。实时荧光定量PCR及2-ΔΔCt法定量分析不同处理组Survivin mRNA的相对表达量。Western blot检测Survivin蛋白表达。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,且随着剂量升高而增加,差异有统计学意义(P<0.05)。流式细胞术检测结果显示,莪术醇β-环糊精包合物处理可使细胞阻滞在G0/G1期与G2/M期,并且促进细胞凋亡。此外,与对照组比较,处理组细胞中Survivin mRNA与蛋白表达水平明显降低(P<0.05)。结论莪术醇β-环糊精包合物能够抑制食管癌细胞TE-1的增殖,促进凋亡,对Survivin的抑制与其抑制食管癌细胞的恶性表型的作用相关。

Objective To investigate the effects and mechanisms of Curcumol β-cyclodextrin Compound （CβC）on the proliferation and apoptosis of esophageal carcinoma cell line TE-I. Methods The CβC was prepared by saturated solution and confirmed by infrared absorption spectroscopy. The effects of CβC （at 25, 50, 100 mg/L）on the proliferation of human esophageal carcinoma cell line TE-1 in vitro was analyzed by MIF assay. The cell cycles and apoptosis were detected by flow cytometer. The relative expression of survivin mRNA was detected by real-time fluorescent quantitative PCR and calculated by the 2^-AACt method. The protein expression of survivin was measured by Western blot. Results Compared with the control group, results of MIF showed that CβC at each dose significantly inhibited the proliferation of TE-1 cells in a dose-dependent manner （ P 〈0.05）. The results of flow cytometry showed that C{3C resulted in the cell cycle arrest at G0/G1and G2/M phase, and promoted the cell apoptosis. Besides, when compared with the control group, the protein and mRNA expressions of survivin obviously decreased in each CβC group （P 〈 0.05）. Conclusions CβC could inhibit the proliferation of esophageal carcinoma cell TE-1 and promote the apoptosis. Its inhibition on the survivin expression was correlated with its inhibition on the malignant phenotypes of esophageal carcinoma cells.