苦参素对肝癌细胞增殖、侵袭和迁移能力的影响及机制

Effects of oxymatrine on proliferation,invasion,and migration of hepatocellular carcinoma cells

目的观察苦参素对肝癌细胞增殖、侵袭和迁移能力的影响,探讨其可能的分子机制。方法取人肝癌细胞株Hep G2,随机分为阴性对照组和苦参素低、中、高浓度组,阴性对照组加入细胞培养液,苦参素低、中、高浓度组分别加入终浓度为1.0、2.0、4.0 mg/m L的苦参素和细胞培养液。采用MTT法测定各组细胞增殖情况,计算细胞增殖抑制率;Transwell小室实验检测各组细胞侵袭和迁移能力;Real-time PCR法检测各组钙黏蛋白(E-cadherin)和CD44 mRNA表达,Western blotting法检测E-cadherin和CD44蛋白表达。结果苦参素高浓度组细胞增殖抑制率较中、低浓度组升高,中浓度组较低浓度组升高。苦参素低、中、高浓度组在细胞侵袭和细胞转移实验中的穿膜细胞数均少于阴性对照组(P均<0.05),苦参素高浓度组少于中、低浓度组,中浓度组少于低浓度组(P均<0.05)。与阴性对照组比较,苦参素低、中、高浓度组E-cadherin mRNA和蛋白表达均升高,CD44 mRNA和蛋白表达降低(P均<0.05)。结论苦参素在体外有抑制人肝癌细胞Hep G2增殖、侵袭和迁移能力的作用,且浓度越高作用越明显;其机制可能与上调E-cadherin表达和下调CD44表达有关。

Objective To investigate the influence and the possible molecular mechanisms of oxymatrine（ OM） on the proliferation,invasion,and metastasis of hepatocellular carcinoma. Methods Human hepatoma cell line Hep G2 was randomly divided into the negative control group,low-dose,medium-dose,and high-dose OM groups. The negative control group was added with the cell culture fluid,besides,the OM groups were added with 1. 0,2. 0,and 4. 0 mg/m L OM and cell culture fluid. The cell proliferation was measured by MTT method and the inhibition rate of cell proliferation was calculated. The invasion and migration of cells in each group were detected by Transwell chamber assay. The expression of Ecadherin and CD44 mRNA in the OM groups was detected by real-time PCR,and Western blotting was used to detect the protein expression of E-cadherin and CD44. Results The cell proliferation inhibition rate of the high-dose OM group was higher than that of the medium-dose and low-dose OM groups,the medium-dose group was higher than that of the low-dose OM group; the number of transmembrane cells in the OM groups was less than that in the negative control group（ all P 0. 05）,the high-dose OM group was less than that in the medium-dose and low-dose OM groups,and the medium-dose group was less than the low-dose group（ all P 〈 0. 05）. The mRNA and protein levels of E-cadherin in the OM groups increased but the mRNA and protein levels of CD44 decreased（ all P 〈 0. 05）. Conclusion OM could inhibit the proliferation,invasion and migration of Hep G2 cells in vitro,and its mechanism may be related to the up-regulation of E-cadherin and down-regulation of CD44 expression.