远华蟾毒精对小鼠乳腺癌细胞侵袭的抑制作用及机制

摘要

Objective To investigate the inhibitory effect and mechanism of Yuanhua toad poison fine on the invasion of mouse breast cancer cells.Methods The cultured mouse breast cancer cells 4T1 were randomly divided into groups A,B,C,and D.Cell in the group A received the conventional culture;cells in the groups B,C,and D were cultured under the hypoxic conditions (at 37 ℃,5％ CO and 95％ N2);while cells in the groups C and D were added with 0.1 and 0.5 mol/L Yuanhua toad poison fine culture.At 0,6,12,and 24 h,the cell migration ability was examined by Scratch test (migration distance);the invasive ability was detected by Transwell invasion test (Number of cells through die);the relative expression levels of HIF-1α,matrix metalloproteinase 9 (MMP-9),phosphoinositide 3-kinase (PI3K),and p-Akt protein were detected by Western blotting.Results The migration distance of each group at different time points was in the following order:group D ＜ group C ＜ group B ＜ group A,the number of transmembrane cells:group D ＜ group C ＜group B ＜ group A,the expression of HIF-1α,MMP-9,p-PI3K,and p-Akt protein:group D ＜ group C ＜ group B ＜group A,and the differences in the above indexes were statistically significant between these two groups (all P ＜ 0.05).Conclusion Yuanhua toad poison fine can decrease the expression of HIF-1α and MMP-9 through the PI3K/AKT pathway,thereby inhibiting the invasion of breast cancer cells.%目的 探讨远华蟾毒精对小鼠乳腺癌细胞侵袭的抑制作用及其机制.方法 将培养好的小鼠乳腺癌细胞4T1随机分为A、B、C、D组.A组常规培养;B、C、D组于缺氧条件下(37℃、5％ CO、95％N2)进行培养,同时C、D组分别加入0.1、0.5 μmol/L远华蟾毒精进行培养.培养0、6、12、24 h采用划痕实验检测细胞迁移能力(迁移距离),用Transwell侵袭实验检测细胞侵袭能力(穿膜细胞数),用Western blotting法检测细胞缺氧诱导因子1α(HIF-1α)、基质金属蛋白酶9(MMP-9)及磷酸化的磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)蛋白表达.结果 不同时点细胞迁移距离D组＜C组＜B组＜A组,穿膜细胞数D组＜C组＜B组＜A组,HIF-1α、MMP-9、p-PI3K、p-Akt蛋白相对表达量D组＜C组＜B组＜A组,以上指标两组间比较差异有统计学意义(P均＜0.05).结论 远华蟾毒精可能通过PI3K/AKT通路降低小鼠乳腺癌细胞HIF-1α、MMP-9的表达,从而抑制细胞侵袭.