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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 inhibitory effect and its mechanism of the Huai’er Purification Cream on the growth of human colon cancer SW480 cells in vitro. Methods: MTT assay was used to detect the growth inhibition of SW480 and explore the optimal concentration of SW480 cells. SW480 cells were cultured for 48 h in vitro and were randomly divided into normoxia group (NC group) Hypoxia group (HC group) and hypoxia falciparum group (HH group). Semi-quantitative RT-PCR was used to detect the expression of vascular endothelial growth factor (VEGF) mRNA in SW480 cells and Western blot was used to detect the protein expression. Results: The inhibitory rate of Huaierqing cream on SW480 cells increased with the increase of the drug concentration. The inhibition rate was the highest (66.7%) at 1mg / ml, which was not significantly different from 5-FU group (10μg / ml). The expression of VEGF mRNA in HH group and HC group was significantly higher than that in NC group (4.71 ± 0.07, 4.54 ± 0.02 and 1.19 ± 0.03, P <0.05), but there was no significant difference between HH group and HC group. The expression of VEGF protein in HC group was significantly lower than that in NC group (0.66 ± 0.03 vs 0.38 ± 0.02, P <0.05). The expression of VEGF protein in HC group was significantly decreased by 0.37 ± 0.03 and 0.66 ± 0.03 (P <0.05), respectively. Conclusion: Huaierqing cream can inhibit the growth of SW480 cells, with the highest inhibition rate at 1 mg / ml. Mechanism for the Huai ear clear cream down SW480 cells VEGF protein expression inhibited tumor.