1. 河南大学附属郑州颐和医院妇科,河南,郑州,450047
2. 郑州大学第一附属医院妇产科,河南,郑州,450052
3. 河南中医药大学人民医院妇产科,河南,郑州,450002
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王永玲, 贾秀改, 朱洁, 等. 粉防己碱调控miR-21表达抑制卵巢癌上皮间质转化的实验研究[J]. 上海中医药杂志, 2020,54(8):71-76.
WANG Yongling, JIA Xiugai, ZHU Jie, et al. An experimental study on inhibitory effect of tetrandrine on regulating expression of miR-21 on epithelial-mesenchymal transition in ovarian cancer[J]. Shanghai Journal of Traditional Chinese Medicine, 2020,54(8):71-76.
王永玲, 贾秀改, 朱洁, 等. 粉防己碱调控miR-21表达抑制卵巢癌上皮间质转化的实验研究[J]. 上海中医药杂志, 2020,54(8):71-76. DOI: 10.16305/j.1007-1334.2020.08.008.
WANG Yongling, JIA Xiugai, ZHU Jie, et al. An experimental study on inhibitory effect of tetrandrine on regulating expression of miR-21 on epithelial-mesenchymal transition in ovarian cancer[J]. Shanghai Journal of Traditional Chinese Medicine, 2020,54(8):71-76. DOI: 10.16305/j.1007-1334.2020.08.008.
目的:研究粉防己碱(TET)调控miR-21表达抑制卵巢癌上皮间质转化(EMT)的作用。 方法:以人卵巢癌细胞(A2780细胞)、人卵巢透明癌细胞(ES-2细胞)和正常人卵巢上皮细胞(IOSE80细胞)作为研究对象,采用MTT法测定0、1.0、2.0、5.0、10.0、20.0 μmol/L的TET对上述3种细胞增殖的影响。实验分为IOSE80细胞对照组、A2780细胞对照组、ES-2细胞对照组、A2780细胞TET组、ES-2细胞TET组,采用实时荧光定量PCR法(RT-qPCR)测定各组细胞miR-21表达水平;迁移实验和侵袭实验考察TET对细胞迁移和侵袭的影响;蛋白印迹法(Western blot)测定各组细胞GSK3β、p-GSK3β、β-catenin、E-cadherin、N-cadherin、Vimentin蛋白表达水平。 结果:①与对照组相比,TET浓度在1.0~20.0 μmol/L时,A2780细胞和ES-2细胞存活率随着TET浓度的增加显著降低(P<0.05);为减少TET药物对细胞的毒性,采用TET对A2780细胞和ES-2细胞的处理浓度分别为5.0 μmol/L和3.0 μmol/L进行后续实验。②与IOSE80细胞组相比,A2780细胞对照组和ES-2细胞对照组miR-21表达水平显著升高(P<0.05);与A2780细胞对照组和ES-2细胞对照组相比,A2780细胞5.0 μmol/L TET组和ES-2细胞3.0 μmol/L TET组miR-21表达水平、迁移能力、侵袭能力、GSK3蛋白磷酸化水平、β-catenin蛋白表达水平、N-cadherin蛋白表达水平、Vimentin蛋白表达水平明显降低(P<0.05),E-cadherin蛋白表达水平明显升高(P<0.05)。 结论:TET可能通过下调miR-21表达水平,从而阻断Wnt/β-catenin信号通路抑制卵巢癌EMT。
Objective:To study the inhibitory effect of tetrandrine(TET) on the level of miR-21 on epithelial-mesenchymal transition(EMT)in ovarian cancer. MethodsHuman ovarian cancer A2780 cells, ES-2 cells and normal human ovarian epithelial IOSE80 cells were used as the research objects, MTT method was used to determine the effects of 0,1.0,2.0,5.0,10.0,20.0 μmol/L TET on the proliferation of three kinds of cells. The experiment was divided into IOSE80 cell control group, A2780 cell control group, ES-2 cell control group, A2780 cell group with TET, ES-2 cell group with TET. The expression level of miR-21 was measured by real-time quantitative PCR(RT-qPCR). The effects of TET on cell migration and invasion were studied by migration and invasion experiments, and the expressions of GSK3β, p-gsk3β, β-Catenin, E-cadherin, N-cadherin and vimentin were measured by Western blot. Results:Compared without TET, the survival rates of A2780 cells and ES-2 cells decreased significantly with the increase of TET concentration(P<0.05)when TET concentration was 1.0~20.0 μmol/L, the concentrations of TET on A2780 cells and ES-2 cells were 5.0 μmol/L and 3.0 μmol/L respectively for the follow-up experiments. Compared with IOSE80 cell comtrol group, the expression level of miR-21 in A2780 cell control group and ES-2 cell control group increased significantly (P<0.05). Compared with the A2780 cell control group and ES-2 cell control group, the miR-21 expression level, migration ability, invasion ability, GSK3 protein phosphorylation level, β-Catenin protein expression level, N-cadherin protein expression level and vimentin protein expression level in the A2780 cell group and ES-2 cell group were significantly reduced(P<0.05), and the expression level of E-cadherin protein increased significantly(P<0.05). Conclusion:TET may block Wnt/β-catenin signaling pathway and inhibit EMT of ovarian cancer by down regulating miR-21 expression.
卵巢癌粉防己碱上皮间质转化miR-21
ovarian cancertetrandrineepithelial-mesenchymal transitionmiR-21
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