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唐恩红, 蔡旺. 茯苓多糖对人宫颈癌HeLa细胞增殖、迁移、促凋亡的影响及其机制[J]. 肿瘤防治研究, 2019, 46(8): 707-713. DOI: 10.3971/j.issn.1000-8578.2019.18.1960
引用本文: 唐恩红, 蔡旺. 茯苓多糖对人宫颈癌HeLa细胞增殖、迁移、促凋亡的影响及其机制[J]. 肿瘤防治研究, 2019, 46(8): 707-713. DOI: 10.3971/j.issn.1000-8578.2019.18.1960
TANG Enhong, CAI Wang. Effects of Pachymaran on Proliferation, Migration and Pro-apoptosis of Human Cervical Carcinoma HeLa Cells and Its Mechanism[J]. Cancer Research on Prevention and Treatment, 2019, 46(8): 707-713. DOI: 10.3971/j.issn.1000-8578.2019.18.1960
Citation: TANG Enhong, CAI Wang. Effects of Pachymaran on Proliferation, Migration and Pro-apoptosis of Human Cervical Carcinoma HeLa Cells and Its Mechanism[J]. Cancer Research on Prevention and Treatment, 2019, 46(8): 707-713. DOI: 10.3971/j.issn.1000-8578.2019.18.1960

茯苓多糖对人宫颈癌HeLa细胞增殖、迁移、促凋亡的影响及其机制

Effects of Pachymaran on Proliferation, Migration and Pro-apoptosis of Human Cervical Carcinoma HeLa Cells and Its Mechanism

  • 摘要:
    目的 探讨茯苓多糖(Pachymaran)对人宫颈癌HeLa细胞增殖、迁移、促凋亡的作用及其相关机制。
    方法 MTT法检测细胞增殖率并筛选出适宜的茯苓多糖低、中、高浓度用于后续实验;不同浓度的茯苓多糖处理细胞后,倒置相差显微镜观察细胞形态学变化;Hoechst 33342染色观察细胞核的变化;平板克隆实验检测细胞克隆形成能力;细胞划痕实验检测细胞迁移能力;流式细胞术检测细胞凋亡率及细胞周期;Western blot法检测凋亡、迁移及ERK通路相关蛋白的表达。
    结果 茯苓多糖浓度对HeLa细胞活力的影响实验得出,取30、40、50 mg/ml茯苓多糖作为低、中、高浓度进行后续实验;中、高浓度茯苓多糖处理细胞后,细胞和细胞核发生显著的凋亡形态学变化,低浓度下形态学变化不显著;不同浓度茯苓多糖均能降低细胞克隆形成能力;降低细胞迁移率(P < 0.05);使细胞凋亡率增加(P < 0.05);使S期细胞减少并阻滞于G2/M期(P < 0.05);Cleaved Caspase 3、Cleaved Caspase 8、Cleaved Caspase 9、Bax表达较对照组明显增多,Bcl-2、MMP-9、VEGFA、p-ERK1/2表达明显减少(P < 0.05)。ERK1/2表达无明显变化。
    结论 茯苓多糖能显著抑制HeLa细胞增殖,并诱导其凋亡。其促凋亡机制可能与下调p-ERK1/2表达,抑制ERK信号通路磷酸化有关。同时,茯苓多糖对抑制HeLa细胞的迁移也有一定的作用。

     

    Abstract:
    Objective To investigate the effects of pachymaran on the proliferation, migration and pro-apoptosis of human cervical carcinoma HeLa cells and their related mechanisms.
    Methods MTT method was used to detect cell proliferation rate, and we selected the suitable low, medium and high concentrations of pachymaran for experiment. After treatment with low, medium and high concentrations of pachymaran, the morphological changes were observed by inverted phase contrast microscope; nuclear changes were observed by Hoechst 33342 staining; flat plate clone formation test was used to detect the ability of cell clone; cell scratch test was used to detect cell migration ability; flow cytometry was used to detect the apoptosis rate and cell cycle distribution. The expression of related proteins of apoptosis, migration and ERK pathway were detected by Western blot.
    Results We selected 30, 40 and 50 mg/ml pachymaran were selected as low, medium and high concentrations in the experiments. After treated with medium and high concentrations of pachymaran, cells and cell nuclei had significant apoptosis morphological changes, but not at low concentration. Different concentrations of pachymaran could reduce the abilities of cell cloning and migration (P < 0.05), increase apoptotic rate (P < 0.05), decreased S phase cells and blocked in G2/M phase (P < 0.05); Cleaved Caspase 3, Cleaved Caspase 8, Cleaved Caspase 9, Bax expression were increased significantly compared with the control group; Bcl-2, MMP-9, VEGFA and p-ERK1/2 expression were decreased significantly (P < 0.05). ERK1/2 expression did not change significantly.
    Conclusion Pachymaran could significantly inhibit the proliferation and induce the apoptosis of HeLa cells. The pro-apoptotic mechanism may be related to the inhibition of phosphorylation of ERK signaling pathway by downregulating the expression of p-ERK1/2. In addition, pathymaran could reduce the migration of HeLa cells to a certain extent.

     

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