花姜酮对MCF7细胞的增殖和迁移与侵袭的影响及其作用机制研究

Effect of Zerumbone on the proliferation,migration and invasion of MCF7 cells and the study of its mechanism

目的研究花姜酮对乳腺癌MCF7细胞增殖、迁移及侵袭的影响及潜在分子机制。方法以四甲基偶氮唑盐(MTT)法检测花姜酮对MCF7细胞增殖的影响;根据MTT结果,将实验分为4组:空白对照组和低、中、高3个剂量实验组。实验组分别给予5,10,20μmol·L-1的花姜酮,空白对照组给予等体积的二甲基亚砜(DMSO),处理48 h。用划痕、Transwell和平板克隆实验测定花姜酮对MCF7细胞迁移、侵袭和克隆形成能力;以免疫印迹法检测相关蛋白表达情况。结果与空白对照组相比,低、中、高剂量花姜酮处理48 h后,MCF7细胞的增殖能力分别降至(89.7±5.4)%,(82.0±6.9)%,(62.7±4.6)%,迁移能力分别降至为(72.7±5.7)%,(55.0±3.9)%,(32.7±4.6)%,侵袭能力降至(77.6±7.8)%,(49.1±4.7)%,(40.8±6.2)%,差异均有统计学意义(均P<0.01)。空白对照组的克隆形成率为(41.4±6.3)%,低、中、高3个剂量实验组依次降至(27.4±4.8)%,(20.4±5.1)%,(5.4±0.8)%,实验组与空白对照组比较,差异均有统计学意义(均P<0.05)。花姜酮处理MCF7细胞后,Ecadherin的表达量随低、中、高3个浓度而梯度上调,但N-cadherin的表达量随浓度而梯度下降,差异均有统计学意义(均P<0.01)。结论花姜酮能够抑制MCF7细胞的增殖、迁移和侵袭,机制可能与促进E-cadherin、抑制N-cadherin的表达有关。

Objective To study the effect of Zerumbone on the proliferation,migration and invasion of MCF7 cells and its underlying mechanism. Methods The effect of Zerumbone on the cell viability of MCF7 cells was determined with 3-（ 4,5-dimethylthiazol-2 yl）-2,5-diphenyltetrazolium bromide（ MTT）. According to the results of MTT,the experiment was divided into four groups： control group（ DMSO） and low,medium,high doses experimental groups（ with corresponding dosage5,10,20 μmol·L^-1 Zerumbone for 48 h）. Wound healing assay,transwell assay and plate colony formation assay were performed to observe the influence of Zerumbone on the migration,invasion and colony forming ability of MCF7 cells. The expression of E-cadherin and N-cadherin was determined by Western blot. Results After treatment of with low,medium and high doses of Zerumbone for 48 h,the proliferation of MCF7 cells in experimental-L,M,H groups was reduced to（ 89.7 ± 5.4） %,（ 82.0 ± 6.9） %,（ 62. 7 ± 4. 6） %; the migration ability of MCF7 cells in the 3 groups was significantly reduced to（ 72.7 ± 5.7） %,（ 55.0 ± 3.9） %,（ 32. 7 ± 4. 6） %; the invasion ability of cells in the 3 groups was reduced to（ 77.6 ± 7.8） %,（ 49. 1 ± 4.7） %,（ 40.8 ± 6.2） %; comparison between experimental groups with the blank control group,the difference of the factors were all statistically significant（ all P〈0.01）. The clone formation rate of the control group was（ 41.4 ± 6.3） %,which was reduced to（ 27.4 ± 4.8） %,（ 20.4 ± 5.1） %,（ 5.4 ± 0.8） % correspondingly in the experimental groups,and there was statistically difference（ all P〈0.05）. The Zerumbone could up-regulate the expression of E-cadherin and down-regulate the expression of N-cadherin in breast cancer cells（ all P〈0.01）. Conclusion Zerumbone can inhibit the proliferation,migration and invasion of MCF7 cells,which may associate with the increase of E-cadherin and the decrease of N-cadherin proteins.