槐耳清膏对结肠癌SW480细胞生长抑制作用及机制研究

Inhibitation and Mechanism of Huaier Cream in Colon Cancer Cells SW480 in Vitro

目的:探讨槐耳清膏对体外培养人结肠癌SW480细胞生长抑制作用及其作用机制。方法:采用甲基噻唑基四唑(MTT)比色法检测槐耳清膏对SW480生长抑制作用并探求最佳作用浓度;体外培养SW480细胞48 h,随机分为常氧组(NC组)、低氧组(HC组)和低氧槐耳组(HH组)。半定量RT-PCR检测SW480细胞血管内皮生长因子(VEGF)mRNA表达水平,Western blot检测蛋白表达水平。结果:槐耳清膏对SW480细胞抑制率随药物浓度增加而上升,1mg/ml时抑制率最大(66.7%),与5-FU组(浓度为10μg/ml)相比无统计学意义。HH组和HC组VEGF mRNA表达均显著高于NC组4.71±0.07,4.54±0.02和1.19±0.03(P<0.05),但HH组与HC组比较差异无统计学意义。HC组VEGF蛋白表达显著高于NC组0.66±0.03和0.38±0.02(P<0.05),HH组较HC组VEGF蛋白表达均显著下降0.37±0.03和0.66±0.03(P<0.05)。结论:槐耳清膏可抑制SW480细胞生长,1 mg/ml时抑制率最大。机制为槐耳清膏下调SW480细胞内VEGF蛋白表达抑制肿瘤。

Objective：To investigate the inhibition and mechanism of Huaier cream in colon cancer cells SW480 in Vitro. Methods：The ability of Huaier cream to inhibit colon cancer cells .SW480 and the top inhabitation ratio were detected by the MTT method. SW480 cells were cultured for .48 hours and randomly divided into normoxic group （NC group）, hypoxia group （HC group） and hypoxia group treated by Huaier （HH group）. Levels of mRNA and protein expression of VEGF were detected by semi-quantitative RT-PCR and .Western blot, respectively. Results： Huaier cream induced a dose dependent inhibition of SW480 cells. The maximum percentage of growth inhibition was 66.7% at a concentration of 1.0 mg/ml, but no significant difference as compared to the positive control（5-FU 10μg/ml）. VEGF mRNA expression levels were significantly higher in both HC group and HH group than in NC group（4.71±0.07, 4.54±0.02 vs. 1.19±0.03, all P〈0.05） ,but not significantly different between HC group and HH group. VEGF protein were over expressed in HC group than NC group （0.66±0.03 vs. 0. 38±0. 02,P〈0. 05）. In HH group, however, VEGF protein was inhibited remarkably compared with HC group （0.37±0.03 vs 0.66±0.03 ,P〈0.05）. Conclusion： Huaier cream can significantly in hibit SW480 cells and reach the top inhabitation ratio at a concentration of 1.0 mg/ml. Huaier cream through down-regulating protein expression of VEGF plays a role in inhibiting tumor.