甘草素通过调控miRNA抑制人黑色素瘤A375细胞的侵袭转移

发布时间：2018-06-15 20:14

本文选题：甘草素 + A细胞　；参考：《南京医科大学学报(自然科学版)》2015年02期

【摘要】：目的 :探讨甘草素对人黑色素瘤A375细胞侵袭转移的抑制作用及与miRNA相关调控机制。方法 :不同浓度甘草素处理24 h后,采用噻唑蓝(MTT)法测定细胞存活情况,确定甘草素对细胞无毒作用剂量;通过划痕试验和Transwell小室迁移、侵袭试验分别观察细胞的非定向迁移力和定向迁移侵袭力;miRNA芯片筛选差异表达的miRNA,q PCR方法验证hsa-miR-4534和hsa-miR-4487的下调表达;Western blot检测PTEN、p-AKT、AKT、MMP2和TIMP2蛋白表达。结果:10~100μmol/L剂量甘草素处理A375细胞24 h对细胞无明显损伤作用,划痕试验、Transwell小室迁移试验、侵袭试验显示甘草素能抑制A375细胞侵袭转移;甘草素可下调hsa-miR-4534和hsa-miR-4487的表达,上调PTEN、TIMP2表达水平,降低p-AKT、MMP2的表达水平。结论:甘草素通过下调hsa-miR-4534和hsa-miR-4487表达,进而上调PTEN和TIMP2靶基因的表达水平,阻碍p-AKT信号通路并降低MMP2蛋白表达,从而发挥抑制人黑色素瘤A375细胞侵袭和转移的作用。
[Abstract]:Objective: To investigate the inhibitory effect of glycyrrhizin on the invasion and metastasis of human melanoma A375 cells and the regulation mechanism related to miRNA. Methods: after 24 h of glycyrrhizin treatment with different concentrations, the cell survival was determined by thiazolium (MTT) method, and the dose of nontoxic action of glycyrrhizin on cells was determined. Through scratch test and Transwell cell migration, invasion test The non directional migration force and directional migration invasiveness of the cells were observed, the differentially expressed miRNA was screened by miRNA chip, and the Q PCR method was used to verify the down regulation of hsa-miR-4534 and hsa-miR-4487; Western blot detected PTEN, p-AKT, AKT, MMP2, and protein expression. Effect, scratch test, Transwell compartment migration test, invasive test showed that glycyrrhizin could inhibit invasion and metastasis of A375 cells; glycyrrhizin could down regulate the expression of hsa-miR-4534 and hsa-miR-4487, up regulation of PTEN, TIMP2 expression, and decrease the expression level of p-AKT, MMP2. Conclusion: glycyrrhizin down regulated the expression of hsa-miR-4534 and hsa-miR-4487 and then up PT. The expression level of the target gene of EN and TIMP2 hinders the p-AKT signaling pathway and reduces the expression of MMP2 protein, thus playing a role in inhibiting the invasion and metastasis of human melanoma A375 cells.
【作者单位】：南京医科大学公共卫生学院营养与食品卫生学系;
【分类号】：R739.5

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