摘要
目的:探讨甘草中黄酮类化合物甘草查尔酮A(licochalcone A,LCA)对小鼠黑色素瘤B16F10细胞株体外迁移能力的影响。方法:取对数生长期B16F10细胞按8 000个/孔接种于96孔板中,磺酰罗丹明B(SRB)法测定LCA(0,5,10,15,20μmol·L-^1)作用24 h对细胞活力影响;取对数生长期B16F10细胞按1×10^5个/孔接种于24孔板中,划痕实验检测LCA(0,5,10,15μmol·L^-1)作用24 h细胞体外迁移能力;取对数生长期B16F10细胞按5×10^5个/瓶接种于细胞培养瓶中,LCA(0,5,10,15μmol·L^-1)作用24 h后,明胶酶谱法检测细胞培养上清基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)活性;ELISA检测MMP-2和MMP-9蛋白含量;实时荧光定量PCR(Q-PCR)和半定量PCR(RT-PCR)法检测MMP-2和MMP-9基因表达水平。结果:5-15μmol·L-1的LCA对小鼠黑色素瘤B16F10细胞作用24 h,对细胞活力没有显著的影响;划痕试验结果说明,LCA可剂量依赖性地降低B16F10细胞的体外迁移能力;明胶酶谱结果显示,LCA可明显抑制B16F10细胞分泌MMP-2和MMP-9的活性;ELISA试验表明,LCA可明显降低MMP-2和MMP-9蛋白表达;Q-PCR和RT-PCR结果显示,LCA可明显降低MMP-2和MMP-9的mRNA基因表达量。结论:甘草查尔酮A能抑制B16F10小鼠黑色素瘤细胞体外转移侵袭作用,其机制可能与抑制MMP-2和MMP-9表达有关。
Objective: The purpose of this study is to investigate the effects of licochalcone A (LCA) on the invasion and metastasis in mouse melanoma B16F10 cells. Method: The cell proliferation rate was evaluated using sulforhodamine B (SRB) method. And the anti-metastasis ability was observed using scratch wound assay. The activated of matrix metal proteinase-2 (MMP-2) and matrix metal proteinase-9 (MMP-9) were evaluated using gelatin zymography. The expression in protein level of MMP-2 and MMP-9 were detected using enzyme-elinked iemmunosorbent essay (ELISA). The mRNA expression of MMP-2 and MMP-9 were assessed using quantitative real-time polymerase chain reaction (PCR) and reverse transcription-PCR. Result: There was no significant effect on proliferation rate after treated with LCA 5-15 μmol·L^-1 in B16F10 cells. LCA attenuated B16F10 cells migration in a concentration-dependent manner. LCA was able to reduce the activities of MMP-2 and MMP-9, down-regulated the expression of MMP-2 and MMP-9 both in mRNA and protein levels. Conclusion: LCA attenuated mouse melanoma B16F10 cells migration by inhibition of MMP-2 and MMP-9 expression.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2015年第18期111-114,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(31471338)