斑蝥素抑制舌癌细胞生长及转移能力

摘 要:【目的】 观察斑蝥素(Cantharidin)对舌癌细胞株Tca8113?CAL-27生长及转移能力的抑制作用?【方法】 舌癌细胞株Tca8113?CAL-27处理组采用斑蝥素处理,通过噻唑蓝(MTT)比色法检测细胞生长能力;通过流式细胞术检测细胞的细胞周期分布;通过实时定量PCR和Western-blot检测p21?细胞周期蛋白依赖激酶1(CDK1)?基质金属蛋白酶-2?9(MMP-2,9)基因表达的变化;通过Transwell小室及划痕实验检测斑蝥素对细胞侵袭和迁移能力的影响?【结果】 斑蝥素抑制舌癌细胞生长,呈剂量和时间依赖性(P﹤0.01)?斑蝥素处理后G2/M期细胞比例上升(P﹤0.01),呈G2/M期阻滞?斑蝥素处理后,抑制G2/M周期转换的p21表达上调(P﹤0.01),而促进G2/M周期转换的CDK1表达下调(P﹤0.01)?斑蝥素可显著抑制舌癌细胞的侵袭能力,处理24 h使Tca8113?CAL-27细胞侵袭力分别下降(93.50 ± 3.32)%(P﹤0.01),(61.33 ± 4.11)%(P﹤0.01)?斑蝥素处理后MMP-2?9 mRNA表达量显著降低?划痕实验显示斑蝥素可显著抑制舌癌细胞的迁移能力(P﹤0.01),处理12 h使Tca8113?CAL-27细胞迁移能力分别下降(57.60 ± 8.98)%(P﹤0.01),(49.94 ± 5.63)%(P﹤0.01)?【结论】 斑蝥素通过阻滞细胞周期抑制舌癌细胞的生长,并通过抑制舌癌细胞迁移和对基质的降解,抑制舌癌细胞转移?

Cantharidin Inhibits Growth and Metastatic Ability of Tongue Cancer Cells

Abstract: 【Objective】 To investigate the effects of cantharidin on the growth and metastatic ability of tongue cancer cell lines Tca8113 and CAL-27. 【Methods】 Cell proliferation was measured by methylthiazol tetrazolium (MTT) assay. Cell cycle was tested by flow cytometry. The expressions of p21, CDK1 (Cyclin dependent kinase 1), matrix metalloproteinase-2, 9 (MMP-2, 9) were detected by real-time PCR and Western blot analysis. Invasion ability was assessed by Transwell invasion assay. Migration ability was evaluated by wound healing assay. 【Results】 Cantharidin inhibited tongue cancer cell growth in a concentration and time-dependent manner (P﹤0.01). After treatment with cantharidin, the ratio of G2/M cells was significantly increased (P﹤0.01). Real-time PCR showed that expression of p21, which represses the G2/M cell cycle transition, was up-regulated (P﹤0.01), while CDK1, which promotes the G2/M cell cycle transition, was down-regulated (P﹤0.01). Treatment with cantharidin for 24 h repressed the invasion ability of Tca8113 and CAL-27 cells by (93.50±3.32) % (P﹤0.01) and (61.33±4.11)% (P﹤0.01), respectively. After cantharidin treatment, the expression of MMP-2, 9 were significantly reduced. Wound healing assay proved that cantharidin repressed the migration of tongue cancer cells by (57.60±8.98) % (P﹤0.01) and (49.94±5.63) %, (P﹤0.01) in Tca8113 and CAL-27 cells respectively. 【Conclusion】 Cantharidin can significantly inhibited the growth of tongue cancer cells through G2/M cell cycle arrest and inhibited the metastatic ability of tongue cancer cells through repressing on the migration and degradation of matrix.