斑螯素抑制胰腺癌细胞株生长、侵袭及其机制

Cantharidin inhibits the growth and invasion ability of pancreatic cancer cells and the mechanism

目的 观察斑蝥素（Cantharidin）对胰腺癌细胞株PANC-1生长及侵袭能力的抑制作用.方法 用斑蝥素处理PANC-1细胞,通过噻唑蓝（MTT）比色法检测细胞生长能力;通过平板克隆形成实验检测细胞克隆形成能力;通过流式细胞术检测细胞的细胞周期分布和凋亡水平;通过transwell侵袭实验检测细胞侵袭能力;通过逆转录-聚合酶链反应（RT-PCR）检测细胞基质金属蛋白酶-2（MMP-2）mRNA表达;通过酶谱法检测细胞MMP-2分泌量.结果 MTT表明随斑瞀素浓度和作用时间增长对胰腺癌细胞PANC-1生长抑制作用增强（P＜0.01）.斑蝥素作用后PANC-1克隆形成能力从未经处理的61.00%降到28.00%,差异有统计学意义（P＜0.01）.10μmol/L斑蝥素48 h处理后细胞G2/M期的比率从未经处理的（16.47±1.66）%增加到（42.71±3.64）%,差异有统计学意义（P＜0.01）,表明斑蝥素使细胞周期阻滞在G2/M期.10 μmol/L斑蝥素作用24 h后,PANC-1细胞凋亡率（24.89±4.80）%较未处理组（7.35±2.44）%明显增加（P＜0.01）,表明斑蝥素诱导细胞凋亡.Transwell侵袭实验显示斑蝥素可显著抑制PANC-1细胞的侵袭能力（P＜0.01）.斑蝥素作用后PANC-1细胞的MMP-2 mRNA表达量及其分泌量显著降低.结论 斑蝥素对胰腺癌细胞的生长和侵袭具有显著的抑制作用.

Objective To investigate the effects of eantharidin on the growth and invasion ability of pancreatic cancer cell line PANC-1. Methods Cell proliferation was measured by methyl thiazol tetrazolium （MTT） assay. Clone formation ability was determined by clony forming assay. Cell cycle and apoptosis were tested by flow cytometry. Invasion ability was assessed by Transwell invasion assay. The expression and secretion level of matrix metalloproteinase-2 （MMP-2） was detected by reverse transcriptionpolymerase chain reaction （RT-PCR） and zymography respectively. Results MTT revealed that cantharidin could inhibit PANC-1 growth in a concentration- and time-dependent manner. After PANC-1 cells were treated with cantharidin, the clone formation ability was significantly decreased from 61.00% to 28.00%（ P 〈0. 01 ）. After PANC-1 cells were treated with 10 μmol/L cantharidin for 48 h, the ratio of G2/M cells was significantly increased from （ 16. 47 ± 1.66 ）% to （42. 71 ± 3.64 ）% （P 〈 0. 01 ）. After PANC-1 cells were treated with 10 μmol/L cantharidin for 24 h, the apoptosis rate in the experimental group （24. 89 ±4. 80） % was significantly higher than in the untreated group （7.35 ± 2. 44） %, （ P 〈 0. 01 ）. These results showed that cantharidin caused G2/M cell-cycle arrest and apoptosis of PANC-1 cells. Transwell invasion assay showed that cantharidin significantly inhibited invasion ability of PANC-1 cells. After PANC-1cells were treated with cantharidin, the expression and secretion of MMP-2 were significantly reduced.Conclusion Cantharidin can significantly inhibit the growth and invasion of pancreatic cancer cells, suggesting cantharidin as an attractive candidate compound for pancreatic cancer therapy.