含酰胺结构的大黄酸衍生物4a对卵巢癌细胞的 迁移、侵袭的影响

Effects of Rhein derivatives 4a containing amide structure on migration and invasion in ovarian cancer cells

目的探讨含酰胺结构的大黄酸衍生物4a(简称衍生物4a,derivative 4a)对卵巢癌SKOV3细胞迁移、侵袭能力的影响及可能作用机制。方法采用分子对接和Western blot检测衍生物4a对Rac1蛋白的调控作用,CCK-8、HE染色、Scratch和Transwell实验分别检测衍生物4a对SKOV3细胞增殖、形态学、迁移和侵袭能力的影响;Western blot检测衍生物4a处理细胞后EMT的标志性蛋白Vimentin、β-cantenin、E-caderin、MMP-2、MMP-9的表达情况。结果衍生物4a能有效与Rac1蛋白结合,结合能明显低于大黄酸;且能下调SKOV3细胞Rac1蛋白的表达。衍生物4a能够明显抑制SKOV3细胞的增殖、侵袭和迁移能力,并诱导细胞产生大量空泡化;衍生物4a可下调MMP-2、MMP-9表达,上调EMT上皮性标志蛋白E-caderin表达及下调Vimentin、β-cantenin的表达。结论衍生物4a能抑制卵巢癌SKOV3细胞增殖、迁移及侵袭,其机制可能是靶向调控Rac1,抑制基质金属蛋白酶分泌,上调EMT过程关键分子E-caderin和下调Vimentin、β-cantenin的表达综合作用的结果。

Aim To investigate the effect of Rhein derivative 4a containing amide structure on migration and invasion in ovarian cancer SKOV3 cells and its possible mechanism.Methods Ovarian cancer SKOV3 cells were used as target cells.Molecular docking and Western blot were used to detect the regulatory effect of derivative 4a on Rac1 protein.CCK8,HE staining,Scratch and Transwell assay were used to detect the effects of derivative 4a on the proliferation,morphology,migration and invasion of SKOV3 cells,respectively.Western blot was employed to determine the expression of matrix metalloproteinases and EMT-related proteins.Results Derivative 4a could effectively bind to Rac1 protein,and the binding energy was-29.10 kcal·mol-1,which was significantly lower than that of Rhein;it also could down-regulate the expression of Rac1 protein in SKOV3 cells.Derivative 4a could significantly inhibit the proliferation,invasion and migration of SKOV3 cells,and induce a large amount of cellular vacuolation;derivative 4a could also down-regulate the expression of MMP-2 and MMP-9,up-regulate the expression of EMT epithelial marker protein E-caderin but down-regulate the expression of vimentin andβ-cantenin.Conclusions Derivative 4a can inhibit the proliferation,migration and invasion of ovarian cancer SKOV3 cells.The mechanism may relate to its targeted regulation of Rac1,thereby inhibiting the secretion of matrix metalloproteinases,up-regulating the expression of key molecule E-caderin and down-regulating the expression of Vimentin andβ-cantenin in EMT process.