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莫艳秀, 姚飞虹, 刘峻彤, 胡紫昂, 李木兰. SP600125对人宫颈癌HeLa细胞增殖和侵袭的影响[J]. 肿瘤防治研究, 2022, 49(4): 304-313. DOI: 10.3971/j.issn.1000-8578.2022.21.0807
引用本文: 莫艳秀, 姚飞虹, 刘峻彤, 胡紫昂, 李木兰. SP600125对人宫颈癌HeLa细胞增殖和侵袭的影响[J]. 肿瘤防治研究, 2022, 49(4): 304-313. DOI: 10.3971/j.issn.1000-8578.2022.21.0807
MO Yanxiu, YAO Feihong, LIU Juntong, HU Ziang, LI Mulan. Effects of SP600125 on Proliferation and Invasion of Human Cervical Cancer HeLa Cells[J]. Cancer Research on Prevention and Treatment, 2022, 49(4): 304-313. DOI: 10.3971/j.issn.1000-8578.2022.21.0807
Citation: MO Yanxiu, YAO Feihong, LIU Juntong, HU Ziang, LI Mulan. Effects of SP600125 on Proliferation and Invasion of Human Cervical Cancer HeLa Cells[J]. Cancer Research on Prevention and Treatment, 2022, 49(4): 304-313. DOI: 10.3971/j.issn.1000-8578.2022.21.0807

SP600125对人宫颈癌HeLa细胞增殖和侵袭的影响

Effects of SP600125 on Proliferation and Invasion of Human Cervical Cancer HeLa Cells

  • 摘要:
    目的 探讨SP600125对人宫颈癌HeLa细胞的增殖周期、凋亡以及侵袭的影响。
    方法 采用CCK-8法检测不同时间点不同浓度的SP600125作用后HeLa细胞的增殖状态。确定20 μmol/L的SP600125用于后续实验。利用平板克隆形成实验检测细胞增殖能力,DAPI染色观察细胞核形态,流式细胞仪检测细胞周期和凋亡,细胞划痕和Transwell法检测细胞迁移和侵袭能力,qRT-PCR和Western blot分别检测SP600125作用不同时间点后各组细胞的p53、Mad2L1和CDC20 mRNA和蛋白水平的变化。
    结果 与对照组(0.1%DMSO)相比,10、20、30、40、50 μmol/L SP600125作用24 h均能使细胞的增殖活性降低。与对照组相比,各SP600125处理组的细胞凋亡率明显增加,且G2/M期细胞比例增加(P < 0.001),而SP600125处理HeLa细胞24和48 h的G0/G1期比例减少(P < 0.001),其细胞的克隆数、迁移和侵袭能力明显下降(P < 0.001);qRT-PCR和Western blot结果显示Mad2L1 mRNA和蛋白表达明显下降(P < 0.05),p53和CDC20 mRNA和蛋白则呈上升趋势(P < 0.01)。
    结论 SP600125可通过上调p53和CDC20以及下调Mad2L1的表达诱导宫颈癌HeLa细胞周期阻滞于G2/M期并促进细胞凋亡来抑制细胞的增殖、迁移和侵袭。

     

    Abstract:
    Objective To investigate the effect of SP600125 on the proliferation, cell cycle, apoptosis and invasion of human cervical cancer HeLa cells.
    Methods CCK-8 method was used to detect the proliferation of HeLa cells treated with different concentrations of SP600125 at different time points. The 20 μmol/L of SP600125 was determined for subsequent experiments. Cell proliferation ability was detected using plate clone formation assay; nuclear morphology was observed by DAPI staining; cell cycle and apoptosis were measured by flow cytometry; cell migration and invasion were detected by cell scratch and Transwell methods; the mRNA and protein levels of p53, Mad2L1 and CDC20 were measured by qRT-PCR and Western blot after SP600125 treatment at different time points.
    Results Compared with control group (0.1%DMSO), cells proliferative activity were reduced by 10, 20, 30, 40 and 50 μmol/L SP600125 treatment for 24h. Compared with control group, the rate of apoptosis was significantly increased in SP600125 treatment groups, and the cell proportion in G2/M phase increased (P < 0.001), while the cell proportion in G0/G1 phases cells was reduced after SP600125 treatment for 24h and 48h (P < 0.001), and the clonal number, migration and invasion ability of HeLa cells also decreased significantly (P < 0.001). qRT-PCR and Western blot results showed a significant decrease in Mad2L1 mRNA and protein expression (P < 0.05) and a significant increase in p53 and CDC20 mRNA and protein expression (P < 0.01).
    Conclusion SP600125 can induce cell cycle arrest of cervical cancer HeLa cells in G2/M phase by upregulating p53 and CDC20 and downregulating Mad2L1 expression, and promote cell apoptosis to inhibit cell proliferation, migration and invasion.

     

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