槐耳清膏对结肠癌细胞生长和迁移的影响及其作用机制

目的中药槐耳清膏具有抗肿瘤效应，但其对结肠癌的作用及机制尚不清楚。本研究旨在探讨槐耳清膏对人结肠癌细胞生长和迁移的影响及作用机制。 方法体外培养人结肠癌细胞系HCT116，采用MTT法检测槐耳清膏对肿瘤细胞生长的抑制作用；采用Boyden趋化小室检测槐耳清膏对肿瘤细胞迁移能力的影响；采用荧光定量PCR法检测药物对MMP-9表达的影响；应用流式细胞技术检测药物对细胞凋亡和细胞周期的影响。 结果槐耳清膏对HCT116细胞生长有明显的抑制作用，该抑制作用具有药物剂量和时间依赖性。与细胞共培养96小时后，槐耳淸膏的抑制作用最强；与对照组相比，10 mg/mL、20 mg/mL和30 mg/mL槐耳淸膏对肿瘤细胞的抑制作用分别为25%(P＝0.025)，42%(P＝0.032)和67%(P＝0.018)。槐耳清膏能明显抑制肿瘤细胞的迁移，11 mg/mL药物的抑制率为63%(P＝0.01)；槐耳清膏(8mg/mL和11mg/mL)处理后的肿瘤细胞中MMP-9的表达量下降(2.73±0.32 vs 4.8±0.36，P＝0.02；1.46±0.25 vs 4.8±0.36，P＝0.004)；槐耳清膏可阻滞细胞停留在S期，但对细胞凋亡无明显影响。 结论体外试验证实槐耳清膏对结肠癌细胞的生长及迁移能力具有抑制作用，其作用机制可能是阻滞细胞停留在S期和下调MMP-9的表达。

Inhibitory effects of Extract of Fungi of Huaier on human colon cancer cells

ObjectiveExtract of Fungi of Huaier(EFH), a traditional Chinese medicine, has been shown to possess anti-tumor effect.However, the role and mechanisms of EFH in colon cancer are still unknown.This study aimed to explore effects of EFH on colon cancer cell growth and migration. MethodsHCT116 cells were cultured in vitro and treated with EFH.MTT method was used to detect growth of HCT116; Boyden chamber was used to test cell migration; Real-time PCR was conducted to analyze the expression of MMP-9; Flow cytometry was applied to detect cell cycle and apoptosis. ResultsCell proliferation assay showed EFH significantly inhibited tumor cell growth compared with the control and inhibition effects were dose and time-dependent.Co-culture with tumor cells for 96 hours, EFH showed the strongest inhibitory effects compared with the control and the inhibitory effects of 10mg/mL, 20mg/mL and 30mg/mL of EFH were 25%(P=0.025), 42%(P=0.032)and 67%(P=0.018)respectively.Moreover EFH(10mg/mL)significantly inhibited the migration of tumor cells with inhibitory rate up to 63%(P=0.01). Expression of MMP-9 in tumor cells treated by EFH(8mg/mL and 11mg/mL)was decreased compared to control(2.73+ 0.32 vs 4.8+ 0.36, P=0.028; 1.46+ 0.25 vs 4.8+ 0.36, P=0.004). In addition, EFH blocked cell cycle progression without triggering cell apoptosis. ConclusionsEFH inhibits colon cancer cell proliferation and migration, and is likely due to down regulation of MMP-9 expression and blockage of cell cycle in S phase.