地高辛对人胃癌SGC7901细胞生长和凋亡的影响

发布时间：2018-09-14 16:07

【摘要】：目的:探究地高辛对人胃癌SGC7901细胞生长和凋亡的影响,并探讨其分子机制。方法:选取人胃癌细胞SGC7901作为研究对象,采用CCK-8法检测地高辛的细胞毒作用并测定IC_(50),采用流式细胞术检测细胞凋亡和细胞周期,采用Western blot法检测c-Src、p-c-Src(Tyr416)、Akt、p-Akt(Ser473)、ERK1/2和p-ERK1/2(Tyr204)的蛋白水平,采用RT-PCR法检测c-Src的mRNA表达水平。结果:随着地高辛浓度逐渐升高,自50nmol/L开始,SGC7901细胞的存活率逐渐降低(P0.05),当地高辛浓度达到500 nmol/L时SGC7901细胞的存活率降至最低(P0.05),地高辛作用24 h的IC_(50)为191.45 nmol/L;200 nmol/L地高辛作用SGC7901细胞24 h后,与对照组相比,细胞凋亡率显著升高(P0.05),G_0/G_1期细胞比例明显升高(P0.05),c-Src、ERK1/2和Akt蛋白的磷酸化水平显著下降(P0.05),c-Src的mRNA和蛋白表达水平均明显降低(P0.05),细胞迁移率降低(P0.05)。结论:地高辛可在体外诱导人胃癌细胞SGC7901的凋亡和G0/G1期阻滞,这可能与其下调c-Src基因表达以及抑制ERK1/2和Akt蛋白磷酸化有关。
[Abstract]:Aim: to investigate the effect of digoxin on the growth and apoptosis of human gastric cancer SGC7901 cells and its molecular mechanism. Methods: the cytotoxic effect of digoxin and IC_ (50) were detected by CCK-8 assay, apoptosis and cell cycle were detected by flow cytometry, and the protein levels of c-Srcnp-c-Src (Tyr416) Aktp-Akt (Ser473) ERK1 / 2 and p-ERK1/2 (Tyr204) were detected by Western blot assay. MRNA expression of c-Src was detected by RT-PCR method. Results: with the increasing of digoxin concentration, The survival rate of SGC7901 cells decreased gradually from the beginning of 50nmol/L (P0.05), and the survival rate of SGC7901 cells decreased to the lowest (P0.05) when the local concentration of goxigenin reached 500 nmol/L. The IC_ (50) of digoxin treated with digoxin for 24 h was 191.45 nmol/L;200 nmol/L digoxin for 24 h, compared with the control group. The rate of apoptosis was significantly increased (P0.05) and the percentage of cells in the G0 / G1 phase was significantly increased (P0.05) the phosphorylation level of c-SrcrK1 / 2 and Akt protein was significantly decreased (P0.05) the expression level of mRNA and protein was significantly decreased (P0.05) and the cell migration rate was decreased (P0.05). Conclusion: digoxin can induce apoptosis and G0/G1 arrest in human gastric cancer cell line SGC7901 in vitro, which may be related to its down-regulation of c-Src gene expression and inhibition of ERK1/2 and Akt protein phosphorylation.
【作者单位】：漯河医学高等专科学校医学生物工程重点实验室;
【基金】：河南省基础与前沿技术研究计划项目(No.142300410061)
【分类号】：R735.2

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