水飞蓟宾对人肝癌HepG2细胞迁移的作用研究

Silybin inhibits migration of human hepatocellular carcinoma cells and its mechanism

目的观察水飞蓟宾对人肝癌(HCC)HepG2细胞迁移的作用。方法采用MTT法观察水飞蓟宾对HepG2细胞的增殖抑制作用,采用细胞划痕实验和Transwell小室法观察水飞蓟宾对HepG2细胞迁移的作用,通过RT-PCR方法检测迁移相关基因Twist、Snail和Slug的表达水平;采用单因素方差分析方法比较不同浓度水飞蓟宾对细胞活力、细胞迁移距离的和迁移细胞个数的影响,并比较各组间Twist、Snail和Slug的转率。结果 MTT检测结果显示不同浓度(0,30,60,120,240,480μg/ml)水飞蓟宾可以不同程度地抑制HepG2细胞增殖,同时呈现剂量依赖和时间依赖(P<0.05);与对照组相比,HepG2细胞在划痕24 h后,水飞蓟宾组(60,120,240μg/ml)的迁移距离分别为(1.50±0.24)cm(P=0.046)、(1.20±0.33)cm(P=0.037)和(1.05±0.24)cm(P=0.029);Transwell实验中水飞蓟宾组(60,120和240μg/ml)细胞迁移个数分别为(100.00±4.25)个、(30.00±5.34)个和(6.00±2.28)个(P均<0.05);PCR结果显示,水飞蓟宾不同程度的降低HepG2细胞Snail、Slug和Twist基因的转率。结论水飞蓟宾可抑制人HCC HepG2细胞的迁移。

Objective To observe the effect of silybinin on the migration of HepG2 cells. Methods MTT assay was used to determine the effect of silybinin on cell proliferation and viability of HepG2 cells. Cell scratch and Transwell assay were used to evaluate the effect of silybinin on migration of HepG2 cells, mRNA transcription level of Twist, Snail and Slug genes was qualified by real-time quantitative PCR. The viability of HepG2 ceils, migration distance, the numbers of migrated cells and gene expression levels of Twist, Snail and Slug were compared among four groups by One- way ANOVA. P 〈 0.05 was considered statistically significant. Results MTT assay showed that silybinin （0, 30, 60, 120, 240, 480 μg/ml） treatment could significantly decrease viability of HepG2 cells in dose- and time- dependent manners（P 〈 0.05）. The migration distances of silybinin group（60, 120 and 240 μg/ml ） were （1.50±0.24） cm （P = 0.046）, （1.20±0.33） cm （P = 0.037 ）and （1.05 ± 0.24）cm （P = 0.029）respectively. Then, we examined the numbers of migrated cells were 100.00 ± 4.25, 30.00± 5.34 and 6.00± 2.28 （P 〈 0.05 ） respectively. Gene expression levels of Twist, Snail and Slug were inhibited in HepG2 ceils （P 〈 0.05）. Conclusion Silybinin could inhibit the migration of HepG2 cells.