隐丹参酮调控Wnt/β-catenin 信号通路对胃癌细胞抑制作用的研究
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R285.5

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国家自然科学青年基金项目(81803776)


A Study on Inhibitory Effect on Gastric Cancer Cells Through Cryptotanshinone Regulating Wnt/β-Catenin Signaling Pathway
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    摘要:

    目的:探究隐丹参酮调控Wnt/β-连环蛋白(β-catenin) 信号通路对胃癌细胞的抑制作用。方法:将人胃癌BGC-823 细胞株分为对照组,隐丹参酮低、中、高剂量组(以下简称低、中、高剂量组),高剂量+SKL2001 组,高剂量+XAV939 组。低、中、高剂量组在培养过程中加入20、40、80 μmol/L 的隐丹参酮;高剂量+SKL2001 组中加入80 μmol/L 隐丹参酮与20 μmol/L SKL2001;高剂量+XAV939 组中加入80 μmol/L 的隐丹参酮与1 μmol/L 的XAV939。采用MTT 法、Transwell 小室法、细胞划痕实验分别检测细胞的增殖能力及迁移侵袭能力。采用RT-PCR 与蛋白免疫印迹法检测细胞中蓬乱蛋白DSH 同源物2(Dvl2)、糖原合成酶激酶-3(GSK-3β)、β-catenin 和G1/S-特异性周期蛋白-D1(Cyclin D1) 表达水平。结果:与对照组比较,低、中、高剂量组处理24 h、48 h、72 h 后细胞增殖抑制率、GSK-3β 蛋白及mRNA 表达上升(P<0.05),Dvl2、β-catenin、Cyclin D1 蛋白及mRNA 表达降低(P<0.05),且呈剂量依赖性(P<0.05)。细胞划痕48 h 后,低、中、高剂量组细胞迁移能力、过膜细胞数及划痕愈合率均降低(P<0.05),且随着剂量增加而降低(P<0.05);与高剂量组比较,高剂量+SKL2001 组迁移能力、过膜细胞数及划痕愈合率均增高(P<0.05);与高剂量+SKL2001 组比较,高剂量+XAV939 组迁移能力、过膜细胞数及划痕愈合率均降低(P<0.05)。结论:隐丹参酮能通过抑制Wnt/β-catenin 信号通路的活性对胃癌细胞起抑制作用。

    Abstract:

    Abstract:Objective: To observe the inhibitory effect on gastric cancer cells through cryptotanshinone regulating Wnt/β- catenin signaling pathway. Methods: Human gastric cancer BGC- 823 cells were divided into three groups: the control group,the low-dose cryptotanshinone group,the medium-dose cryptotanshinone group,the high-dose cryptotanshinone group(shorthand for the low,medium and high dose groups),the high-dose +SKL2001 group and the high-dose +XAV939 group. The low,medium and high dose groups were applied with 20,40 and 80 μmol/L cryptotanshinone during the culture process;the high-dose +SKL2001 group was added with 80 μmol/L cryptotanshinone and 20 μmol/L SKL2001;the highdose +XAV939 group was added with 80 μmol/L cryptotanshinone and 1 μmol/L XAV939. The MTT assay,Transwell assay and cell scratch assay were used to detect the proliferation,migration and invasion of cells. The RT-PCR and protein western blot were used to detect the expression levels of Dishevelled-2(Dvl2),Glycogen Synthase Kinase-3β(GSK-3β),β-catenin and Cyclin D1 in cells. Results:Compared with those in the control group,the cell proliferation inhibition rate,levels of GSK- 3β protein and mRNA expression were increased in the the low, medium and high dose groups 24, 48 and 72 h after intervention(P<0.05), and the levels of Dvl2, β- catenin, Cyclin D1 protein and mRNA expression were decreased(P< 0.05), and the dose- dependent feature was shown(P<0.05). Forty- eight hours after cell scratch, the migration ability of cells,the number of transmembrane cells and the healing rate of the scratch were all decreased in the the low,medium and high dose groups(P<0.05), and they decreased as the dose increased(P<0.05). Compared with those in the high dose group, the migration ability, the number of transmembrane cells and the healing rate of the scratch in the high- dose + SKL2001 group were all increased(P<0.05);compared with those in the high-dose +SKL2001 group,the above indexes in the high-dose +XAV939 group were all decreased(P<0.05). Conclusion:Cryptotanshinone can inhibit gastric cancer cells by inhibiting the activity of Wnt/β-catenin signaling pathway.

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马丽娟,王锡恩,刘培,陈蜜,张静,彭敏.隐丹参酮调控Wnt/β-catenin 信号通路对胃癌细胞抑制作用的研究[J].新中医,2020,52(21):1-5

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  • 在线发布日期: 2020-11-05
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