苦参碱对HCCLM3人高转移性肝癌细胞体外转移的作用及机制研究

Study on the Effect and Mechanism of Matrine on the Metastasis of HCCLM3 Human Highly Metastatic Liver Cancer Cells in vitro

目的:探讨苦参碱对HCCLM3人高转移肝癌细胞体外转移的作用及其机制。方法:CCK-8检测不同浓度的苦参碱对HCCLM3癌细胞的增殖作用,选取不具有直接抑制细胞增殖效应的药物浓度进行细胞划痕实验,研究苦参碱对HCCLM3癌细胞迁移的影响;Transwell实验研究苦参碱对HCCLM3癌细胞侵袭的抑制影响;实时定量PCR(RT-PCR)和WesternBlot实验研究苦参碱对HCCLM3癌细胞E-钙粘蛋白(E-cadherin)、N-钙粘蛋白(N-cadherin)、波形蛋白(Vimentin)、基质金属蛋白酶(Matrix Metalloprotinase,MMP)-2和MMP-9因子mRNA和蛋白的表达。结果:苦参碱4、8、16 mg·mL-1剂量对HCCLM3癌细胞不产生直接抑制作用(P>0.05);苦参碱4、8、16 mg·mL-1剂量能够抑制HCCLM3癌细胞的迁移和侵袭,且与苦参碱0 mg·mL-1组相比,差异具有统计学意义(P<0.05);苦参碱4、8、16 mg·mL-1剂量能够减少HCCLM3癌细胞的N-cadherin、Vimentin、MMP-2和MMP-9因子mRNA和蛋白的表达,并增加E-cadherin mRNA和蛋白的表达。结论:苦参碱能够抑制HCCLM3癌细胞的转移和侵袭,其机制可能与抑制细胞上皮间质转化(Epithelial-mesenchymal transition,EMT)相关蛋白以及MMP-2和MMP-9有关。

Objcetive:To investigate the effect and mechanism of matrine on the metastasis of human highly metastatic liver cancer cells HCCLM3.Methods:CCK-8 was used to detect the proliferation effects of different concentrations of matrine on HCCLM3 cells,and the concentrations that did not directly inhibit cell proliferation were selected for cell scratch experiments to study the effects of matrine on the migration of HCCLM3 cells;Transwell experiments were used to study the inhibitory effects of matrine on HCCLM3 cell invasion;Real-time quantitative PCR(RT-PCR)and Western Blot experiments were used to study matrine on the mRNA and protein expression of E-cadherin,N-cadherin,vimentin,matrix metalloprotinase(MMP)-2 and MMP-9 in HCCLM3 cells.Results:Matrine at 4,8 and 16 mg·mL^(-1) did not inhibit the proliferation of HCCLM3(P>0.05);Matrine at 4,8 and 16 mg·mL^(-1) could inhibit the migration and invasion of HCCLM3 cells.Matrine at 4,8 and 16 mg·mL^(-1) reduced the mRNA and protein expression of N-cadherin,vgimentin,MMP-2 and MMP-9 in HCCLM3 cells,while increased the mRNA and protein expression of E-cadherin.Conclusion:Matrine can inhibit the metastasis and invasion of human highly metastatic liver cancer cells HCCLM3,and its mechanism may be the inhibition of MMP-2,MMP-9 and epithelial-mesenchymal transition(EMT)related proteins.