木犀草素对人舌鳞癌细胞侵袭和迁移的影响

Influence of luteolin on the invasion and migration of an human tongue squamous carcinoma cell line

目的探讨木犀草素(Luteolin)对人舌鳞状细胞癌细胞SCC15侵袭和迁移能力的影响。方法采用浓度梯度的木犀草素药物(5、10、15、20、40、60μg/mL),分别处理SCC15细胞24 h、48 h、72 h,通过MTT实验检测木犀草素对SCC15细胞增殖能力的影响;采用浓度梯度的木犀草素药物(1、5、10μg/mL)分别处理SCC15细胞,通过划痕实验检测木犀草素对SCC15细胞迁移能力的影响;采用浓度梯度的木犀草素药物(5、10μg/mL)分别处理SCC15细胞24 h,通过Transwell实验检测木犀草素对SCC15细胞侵袭和迁移能力的影响;采用浓度梯度的木犀草素药物(10、20、40μg/mL)分别处理SCC15细胞24 h,通过明胶酶谱法检测木犀草素对SCC15细胞上清液中基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)活性的变化。结果 MTT实验表明木犀草素对SCC15细胞增殖具有明显的抑制作用(P<0.01),并呈现明显的剂量—效应和时间—效应关系;划痕实验、Transwell小室迁移及侵袭实验表明实验组细胞的迁移和侵袭能力均明显低于对照组,迁移实验3组穿膜细胞数分别为(340.00±22.94)个、(52.67±6.94)个和(6.57±0.80)个;侵袭实验3组穿膜细胞数分别为(85.67±5.18)个、(39.67±4.63)个和(2.67±0.29)个(P<0.01);明胶酶谱显示木犀草素能明显降低MMP-2、MMP-9的酶活性,呈剂量依赖性(P<0.01)。结论木犀草素可通过抑制明胶酶MMP-2、MMP-9的分泌从而抑制SCC15细胞的侵袭和迁移。

Objective To investigate the effects of luteolin on the invasion and migration of the human tongue squamous carcinoma cell line SCCl5. Methods SCC15 cells were treated with various concentrations of luteolin（5, 10, 15,20, 40 and 60 μg/mL） for 24, 48 and 72 h. The MTT assay was then carried out to estimate the proliferation of SCC15 cells treated with various concentrations of luteolin. SCC15 cells were treated with various concentrations of luteolin（1,5 and 10 μg/mL）, and the migration of SCC15 cells was examined in wound healing assays. SCC15 cells were treated with various concentrations of luteolin（5 and 10 μg/mL） for 24 h, and the migration and invasion of the cells were examined in Transwell migration/invasion assays. SCC15 cells were treated with various concentrations of luteolin（10, 20 and 40 μg/mL） for 24 h, and the conditioned medium was collected. The levels of the gelatinases matrix metalloproteinases-2 and-9（MMP-2, MMP-9） in the conditional medium were detected by gelatin zymography assays. Results The MTT assay showed that luteolin had a substantial inhibitory effect on the proliferation of SCC15 cells in a concentrationand time-dependent manner（P〈0.01）. The migration, invasion and proliferation of the SCCl5 cell lines were significantly lower after treatment with luteolin than in the control. The numbers of migrating and invading SCCl5 cells were340.00 ± 22.94, 52.67 ± 6.94 and 6.57 ± 0.80 versus 85.67 ± 5.18, 39.67 ± 4.63 and 2.67 ± 0.29, respectively（P〈0.01）. The enzyme activities of MMP-2 and MMP-9 decreased significantly in response to luteolin treatment in a concentration-dependent manner（P〈0.01）. Conclusion Luteolin inhibited the invasion and migration of SCC15 cells by reducing the activities of MMP-2 and MMP-9.