紫云英苷通过上调PHD2抑制缺氧诱导的卵巢癌细胞增殖和侵袭

doi:10.11904/j.issn.1002-3070.2019.05.005

摘要/Abstract

摘要： 目的探索紫云英苷(ATG)抑制卵巢癌细胞HIF-1α表达及细胞增殖和侵袭的作用及机制。方法分别给予缺氧条件培养的OVCAR-8和SK-OV-3细胞ATG(ATG组)或ATG联合PHD2抑制剂IOX2(ATG+IOX2组)处理24h,并以仅缺氧培养24h的细胞为空白对照。采用Western blot和实时荧光定量PCR检测各组细胞HIF-1α、PHD2、PCNA、MMP-2和MMP-9蛋白和mRNA表达变化;以正常条件培养的细胞为阴性对照,采用缺氧试剂盒检测ATG处理后细胞缺氧状态变化;采用Transwell实验检测各组细胞侵袭能力;采用CCK-8试剂盒检测各组细胞增殖活力。分别以梯度浓度的ATG联合梯度浓度的卡铂或顺铂处理OVCAR-8和SK-OV-3细胞72h后采用CCK-8试剂盒检测细胞增殖活力,并进行联合效应分析。结果缺氧培养条件下,与空白对照组相比,ATG组OVCAR-8和SK-OV-3细胞PHD2蛋白及mRNA表达均升高,细胞增殖和侵袭能力均降低,HIF-1α蛋白表达均减少,PCNA、MMP-2和MMP-9蛋白及mRNA表达均降低,但HIF-1α的mRNA水平以及细胞缺氧状态无明显变化;而与ATG组相比,ATG+IOX2组OVCAR-8和SK-OV-3细胞增殖和侵袭能力均增加,PCNA、MMP-2、MMP-9蛋白和mRNA表达及HIF-1α蛋白表达均升高,但HIF-1α的mRNA水平无明显变化。此外,ATG可明显增强OVCAR-8和SK-OV-3细胞对顺铂和卡铂的敏感性,具有较好的联合效应。结论 ATG可通过上调PHD2表达,促进缺氧诱导的HIF-1α降解,进而抑制卵巢癌细胞增殖和侵袭。

关键词: 紫云英苷, 卵巢癌, 缺氧, HIF-1α, 侵袭

Abstract: Objective The objective of this study was to explore the effect and mechanism of astragaline(ATG)on inhibition of HIF-1α,proliferation and invasion of ovarian cancer cells.Methods OVCAR-8 and SK-OV-3 cells were cultured in hypoxic conditions and treated with ATG(ATG group)or ATG combined with PHD2 inhibitor IOX2(ATG+IOX2 group)for 24h.The cells were just cultured in hypoxic condition for 24 h as a blank control.Western blot and real-time fluorescent quantitative PCR were used to detect the expression of HIF-1α,PHD2,PCNA,MMP-2 and MMP-9 at levels of mRNA and protein in each group.The cells cultured under normal conditions were used as a negative control,and the changes of hypoxic status in cells were detected after ATG treatment by hypoxia kit.Transwell assay was used to detect the invasive ability of cells in each group;CCK-8 kit was used to determine the activity of cell proliferation in each group.OVCAR-8 and SK-OV-3 cells were treated with gradient concentrations of ATG combined with gradient concentrations of carboplatin or cisplatin for 72h,the cell proliferation activity was detected by CCK-8 kit,and combined effect analysis was also performed.Results Under hypoxic conditions,compared with the blank control group,the expression of PHD2 at levels of protein and mRNA in the ATG group were increased,cell proliferation and invasion ability were decreased,and the expression of HIF-1α protein was decreased in OVCAR-8 and SK-OV-3 cells;the expression of PCNA,MMP-2 and MMP-9 at levels of protein and mRNA were decreased;but the expression of HIF-1α mRNA and cell hypoxia status did not change.When compared with the ATG group,the abilities of proliferation and invasion in the ATG+IOX2 group were increased in OVCAR-8 and SK-OV-3 cells;the expression of PCNA,MMP-2 and MMP-9 at levels of protein and mRNA as well as the expression of HIF-1α protein were increased,but the mRNA level of HIF-1α did not change.In addition,ATG could significantly enhance the sensitivity of OVCAR-8 and SK-OV-3 cells to cisplatin and carboplatin,and both combinations showed a good synergistic effect.Conclusion ATG can promote the degradation of HIF-1α by hypoxia via up-regulating PHD2 expression,resulting in inhibiting proliferation and invasion of ovarian cancer cells.

Key words: Astragaline, Ovarian cancer, Hypoxia, HIF-1α, Invasion

中图分类号:

R737.31

宋玲, 付琼. 紫云英苷通过上调PHD2抑制缺氧诱导的卵巢癌细胞增殖和侵袭[J]. 实用肿瘤学杂志, 2019, 33(5): 407-414.

SONG Ling, FU Qiong. Astragalin inhibits hypoxia-induced proliferation and invasion of ovarian cancer cells by up-regulating PHD2[J]. Journal of Practical Oncology, 2019, 33(5): 407-414.

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链接本文: http://journal09.magtechjournal.com/Jwk3_syzlxzz/CN/10.11904/j.issn.1002-3070.2019.05.005

http://journal09.magtechjournal.com/Jwk3_syzlxzz/CN/Y2019/V33/I5/407

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