甘草素通過調(diào)控miRNA抑制人黑色素瘤A375細胞的侵襲轉(zhuǎn)移
發(fā)布時間:2018-06-15 20:14
本文選題:甘草素 + A細胞; 參考:《南京醫(yī)科大學(xué)學(xué)報(自然科學(xué)版)》2015年02期
【摘要】:目的 :探討甘草素對人黑色素瘤A375細胞侵襲轉(zhuǎn)移的抑制作用及與miRNA相關(guān)調(diào)控機制。方法 :不同濃度甘草素處理24 h后,采用噻唑藍(MTT)法測定細胞存活情況,確定甘草素對細胞無毒作用劑量;通過劃痕試驗和Transwell小室遷移、侵襲試驗分別觀察細胞的非定向遷移力和定向遷移侵襲力;miRNA芯片篩選差異表達的miRNA,q PCR方法驗證hsa-miR-4534和hsa-miR-4487的下調(diào)表達;Western blot檢測PTEN、p-AKT、AKT、MMP2和TIMP2蛋白表達。結(jié)果:10~100μmol/L劑量甘草素處理A375細胞24 h對細胞無明顯損傷作用,劃痕試驗、Transwell小室遷移試驗、侵襲試驗顯示甘草素能抑制A375細胞侵襲轉(zhuǎn)移;甘草素可下調(diào)hsa-miR-4534和hsa-miR-4487的表達,上調(diào)PTEN、TIMP2表達水平,降低p-AKT、MMP2的表達水平。結(jié)論:甘草素通過下調(diào)hsa-miR-4534和hsa-miR-4487表達,進而上調(diào)PTEN和TIMP2靶基因的表達水平,阻礙p-AKT信號通路并降低MMP2蛋白表達,從而發(fā)揮抑制人黑色素瘤A375細胞侵襲和轉(zhuǎn)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.
【作者單位】: 南京醫(yī)科大學(xué)公共衛(wèi)生學(xué)院營養(yǎng)與食品衛(wèi)生學(xué)系;
【分類號】:R739.5
【參考文獻】
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