牡荆葡基黄酮抑制自噬诱导非小细胞肺癌A549 细胞凋亡及对移植瘤裸鼠存活的影响
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R734.2

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河南省科技攻关计划项目(172102310585)


Study on Effect of Vitex Gluconyl Flavonoids on Inhibiting Autophagy- Induced Apoptosis of Non-Small Cell Lung Cancer A549 Cells and its Effect on Survival of Nude Mice with Transplanted Tumor
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    摘要:

    目的:探讨牡荆葡基黄酮对人非小细胞肺癌A549 细胞凋亡和移植瘤裸鼠存活的影响。方法:体外实验:用不同浓度(10,20,40 μmol/L) 的牡荆葡基黄酮作用于肺癌A549 细胞。CCK8 法检测细胞增殖;流式细胞技术检测细胞凋亡;蛋白印迹法检测Ki67,PCNA,Caspase-3,Caspase-9,P62,Beclin1,LC3Ⅱ/LC3Ⅰ,ATG5 的蛋白表达及JNK1/2,ERK1/2 的磷酸化情况。体内实验:皮下注射A549 细胞建立移植瘤模型,分别腹腔注射0.5,1,2 mg/kg 牡荆葡基黄酮,于第30 天绘制存活曲线,免疫组化检测组织中Ki67 和Caspase-3 的表达情况。结果:牡荆葡基黄酮能抑制A549 细胞存活和Ki67、PCNA 蛋白表达;促进细胞凋亡和Caspase-3、Caspase-9 蛋白表达;下调自噬标记物Beclin1、LC3Ⅱ/LC3Ⅰ、ATG5,上调P62 蛋白表达;增加JNK1/2,减少ERK1/2 的磷酸化水平;还能增加移植瘤裸鼠存活率,下调Ki67 而上调Caspase-3 的表达水平。结论:牡荆葡基黄酮能通过抑制细胞自噬而诱导A549 细胞凋亡,同时增加移植瘤裸鼠的存活率。

    Abstract:

    Abstract: Objective: To observe the effect of vitex gluconyl flavonoids on inhibiting autophagy- induced apoptosis of non- small cell lung cancer A549 cells and its effect on survival of nude mice with transplanted tumor. Methods: In vitro experiment,different concentrations(10 μmol/L,20 μmol/L,40 μmol/L) of vitex gluconyl flavonoids were used to treat lung cancer A549 cells. Cell proliferation was detected by CCK8 assay;cell apoptosis was detected by flow cytometry;Western Blotting was used to detect the protein expressions of Ki67,PCNA,Caspase-3,Caspase-9,P62,Beclin1,LC3Ⅱ/LC3Ⅰ, ATG5 and the phosphorylation of JNK1/2 and ERK1/2. In vivo experiment,the transplanted tumor models were established by subcutaneous injection of A549 cells, and they were intraperitoneal injected with 0.5 mg/kg, 1 mg/kg, 2 mg/kg Vitex gluconoids,respectively. The survival curve was plotted on Day 30,and the expression of Ki67 and Caspase- 3 in tissues was detected by immunohistochemistry. Results:Vitex gluconyl flavonoids could inhibit the survival of A549 cells,decrease the expression of Ki67 and PCNA proteins,promote cell apoptosis and expression of Caspase- 3 and Caspase- 9 proteins, down-regulate utophagy markers Beclin1,LC3Ⅱ/LC3Ⅰ and ATG5,up-regulate P62 protein expression,increase JNK1/2 and decrease the phosphorylation level of ERK1/2. It also increased the survival rate,down- regulated Ki67 expression and up- regulated the expression of Caspase- 3 in nude mice with transplanted tumor. Conclusion: Vitex glucomylflavone can induce apoptosis of A549 cells by inhibiting autophagy,and increase the survival rate of nude mice with transplanted tumor.

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钟春蕾,黄国涛,张志强.牡荆葡基黄酮抑制自噬诱导非小细胞肺癌A549 细胞凋亡及对移植瘤裸鼠存活的影响[J].新中医,2021,53(17):141-146

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  • 在线发布日期: 2021-09-09
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