青蒿琥酯对结肠癌细胞增殖、凋亡及β-catenin表达的影响

The Effect of Artesunate on Proliferation,Apoptosis and β-catenin Expression in HT29 Cells

目的探讨青蒿琥酯对结肠癌细胞增殖、凋亡及β-catenin表达的影响。方法取对数生长期的结肠癌HT-29细胞,用12.5μg·m L^(-1)、25μg·m L^(-1)、50μg·m L^(-1)的青蒿琥酯分别培养24 h、48 h和72 h。MTT检测细胞增殖;Annexin V/PI双染检测细胞凋亡率;流式细胞术检测细胞周期分布;Western blot检测β-catenin蛋白表达。结果 MTT结果显示,青蒿琥酯能明显抑制结肠癌HT-29细胞增殖,与对照组(0μg·m L^(-1))相比,差异具有统计学意义(P<0.01);Annexin V/PI检测结果表明,细胞的凋亡率随着青蒿琥酯浓度的增高而逐渐增高,明显高于对照组(P<0.05);流式细胞仪检测细胞周期变化的结果显示,HT-29细胞G0/G1期比例与青蒿琥酯浓度呈反比(P<0.05),G2/M及S期比例逐渐增加(P<0.05)。Western blot检测结果证实,青蒿琥酯处理细胞后β-catenin表达随药物浓度升高而降低(P<0.05)。结论青蒿琥酯可抑制结肠癌细胞增殖,促进细胞凋亡,下调β-catenin的表达,有望成为临床治疗肿瘤的新型药物。

Objective To explore the effects of artesunate on proliferation, apoptosis and β-catenin expression in HT29 cells. Meth- ods The logarithmic phase of human colon cancer cells （HT-29 cells） were used and were treated with different concentration of artesunate （including 12.5 μg.mL-1, 25 μg.mL-l, 50 μg-mL-1） for 24 h, 48 h and 72 h respectively. MTT method was used to detect cells prolifera- tion; AnnexinV/PI double straining was used to detect cells apoptosis rate; Flow cytometry was applied to detect the cell cycle distribution; and Western blot was applied to measure the β-catenin protein expression. Results The results of MTT assay indicated that the artesunate could control the proliferation of HT-29 cells compared with the control group （0 μg.mL-1） （P〈0.01）; AnnexinV/PI test results showed that HT-29 cells apoptosis was improved with the increase of the concentration of artesunate, and clearly higher than the control group （P 〈 0.05）. With the increase of artesunate concentration, flow cytometry results showed that HT-29 cells G0/G1 phase ratio gradually reduced （P 〈 0.05）, while G2/M and S phase ratios were gradually increased （P〈0.05）. Western blot confirmed that the expression of β-catenin was decreased along with increased drug concentration after artesunate treatment of cells. Conclusion Artesunate can inhibit the proliferation of HT29 cells, promote apoptosis and reduce β-catenin expression, which is expected to be used for new drugs in the clinical treatment of tumor.