毛兰素对人肺癌A549细胞迁移、侵袭、凋亡的影响及可能机制研究-现代医学

毛兰素对人肺癌A549细胞迁移、侵袭、凋亡的影响及可能机制研究

单位：1. 中国人民解放军联勤保障部队第九二八医院肿瘤内科, 海南海口 570100;
2. 海南省肿瘤医院胃食管科, 海南海口 570100

分类号：R734.2

出版年·卷·期（页码）：2022·50·第八期（968-973）

摘要：

目的：探讨毛兰素(erianin)对人肺癌A549细胞增殖、迁移和凋亡的影响，并阐述可能机制。方法：将人肺癌A549细胞分为对照组、40 nmol·L^-1毛兰素组、80 nmol·L^-1毛兰素组、160 nmol·L^-1毛兰素组和阳性对照组，分别采用含0、40、80、160 nmol·L^-1毛兰素和20 μl·ml^-1顺铂的RPMI-1640培养基常规培养。采用磺酰罗丹明B(SRB)法检测各组A549细胞增殖率；划痕实验检测各组A549细胞迁移；吖啶橙/嗅化乙锭(AO/EB)染色法检测各组A549细胞凋亡；免疫荧光法检测各组A549细胞中Caspase-3、MMP-2和MMP-9蛋白表达；Western blot法检测各组A549细胞中Cl-PARP、Bcl-2、Survivin、Cl-Caspase-9、Cl-Caspase-8和 Cl-Caspase-3蛋白表达水平。结果：与对照组比较，40、80、160 nmol·L^-1毛兰素组A549细胞增殖率逐渐下降；与对照组比较，不同浓度毛兰素组细胞迁移距离增加，呈浓度依赖性；培养48 h后，与对照组比较，不同浓度毛兰素组凋亡细胞数逐渐增多，呈浓度依赖性；免疫荧光观察结果，与对照组比较，不同浓度毛兰素组Caspase-3阳性蛋白表达逐渐升高，而MMP-2和MMP-9阳性蛋白表达逐渐下降；Western blot法检测结果，与对照组比较，40、80、160 nmol·L^-1毛兰素组细胞中Cl-PARP、Cl-Caspase-9、Cl-Caspase-8和 Cl-Caspase-3蛋白表水平逐渐升高，而Bcl-2和survivin蛋白表达水平逐渐下降，呈浓度依赖性。结论：毛兰素能抑制人肺癌A549细胞迁移、侵袭，促进凋亡，其机制可能与上调Bcl-2蛋白表达，活化Caspase-3，从而促进A549细胞凋亡有关。

Objective: To investigate the effects of erianin on proliferation,migration and apoptosis of human lung cancer A549 cells,and to elaborate the possible mechanism.Methods: Human lung cancer A549 cells were divided into control group,40 nmol·L^-1 erianin group,80 nmol·L^-1 erianin group,160 nmol·L^-1 erianin group and positive control group.RPMI-1640 medium containing 0,40,80 and 160 nmol·L^-1 erianin was used for routine culture.The proliferation rate of A549 cells in each group was detected by SRB.Apoptosis of A549 cells in each group was detected by AO/EB staining.The migration of A549 cells in each group was detected using wound healing assay.The expression levels of Caspase-3,MMP-2 and MMP-9 in A549 cells were detected with immunofluorescence.The expression levels of Caspase-3,MMP-2,MMP-9,Cl-Caspase-9,Cl-Caspase-8 and Cl-Caspase-3 in A549 cells were detected using Western blot.Results: Compared with the control group,the proliferation rate of A549 cells in the 40,80,160 nmol·L^-1 erianin groups decreased gradually;after 48 h of culture,compared with the control group,the number of apoptotic cells in the erianin groups with different concentrations gradually increased in a concentration-dependent manner.Immunofluorescence showed that compared with the control group,the expression of Caspase-3 positive protein in the erianin groups with different concentrations gradually increased,while the expression of MMP-2 and MMP-9 positive proteins gradually decreased.Western blot showed that compared with control group,the expression of Cl-PARP,Cl-Caspase-9,Cl-Caspase-8 and Cl-Caspase-3 protein in the cells of 40,80,160 nmol·L^-1 erianin group increased gradually,while the expression of Bcl-2 and survivin protein decreased gradually in a concentration-dependent manner.Conclusion: Erianin can inhibit the migration and invasion of human lung cancer A549 cells and promote their apoptosis.Its mechanism may be related to up regulating the expression of Bcl-2 protein and activating Caspase-3,so as to promote the apoptosis of A549 cells.

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