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Received:November 03, 2021 Published Online:May 20, 2022
Received:November 03, 2021 Published Online:May 20, 2022
中文摘要: 目的 探讨雷公藤红素对非小细胞肺癌A549细胞增殖、侵袭和迁移的作用及其机制。方法 不同浓度雷公藤红素处理非小细胞肺癌A549细胞后,用CCK-8实验检测细胞增殖抑制作用及半数抑制率(IC50);用显微镜观察A549细胞形态;用平板克隆实验检测A549细胞集落形成能力;用Annexin V/PI双染流式细胞术检测A549细胞凋亡率;用Transwell实验和划痕实验检测A549细胞侵袭和迁移能力;用Western blot实验检测环氧合酶-2(COX-2)、基质金属蛋白酶(MMP)-2、MMP-9、p-p65、p65蛋白表达;用RT-qPCR实验检测COX-2 mRNA、MMP-2 mRNA、MMP-9 mRNA表达。结果 雷公藤红素可以抑制A549细胞的细胞活力(P<0.05),48 h IC50为(2.607±0.163)μmol/L。A549细胞经雷公藤红素处理后,细胞数目显著减少,细胞皱缩变圆,细胞贴壁能力减弱,细胞间距变大,细胞丝状伪足减少。雷公藤红素能明显抑制A549细胞的集落形成能力,诱导A549细胞凋亡,抑制A549细胞侵袭迁移能力,抑制A549细胞中COX-2蛋白、MMP-2蛋白、MMP-9蛋白、p-p65/p65、COX-2 mRNA、MMP-2 mRNA、MMP-9 mRNA的表达,差异有统计学意义(P<0.05)。结论 雷公藤红素能够抑制非小细胞肺癌A549细胞增殖、侵袭和迁移能力,可能与其通过核因子(NF)-κB信号下调COX-2、MMP-2、MMP-9蛋白和mRNA的表达有关。
Abstract:ObjectiveTo explore the effect and mechanism of celastrol on the proliferation, invasion and migration of non-small cell lung cancer (NSCLC)A549 cells. Methods After different concentrations of celastrol were used to treat NSCLC A549 cells, CCK-8 assay was used to detect cell proliferation inhibition and half inhibition rate (IC50); the morphology of A549 cells was observed by microscope; the colony formation ability of A549 cells was detected by plate cloning assay; the apoptosis rate of A549 cells was detected by Annexin V/PI double-staining flow cytometry; the invasion and migration ability of A549 cells were detected by Transwell assay and scratch assay; western blot was used to detect the changes of COX-2, MMP-2, MMP-9, p-p65, p65 protein expressions; RT-qPCR experiments were used to detect the changes of COX-2 mRNA, MMP-2 mRNA, MMP-9 mRNA expression. Results Celastrol inhibited the cell viability of A549 cells (P<0.05), and the 48 h IC50 was (2.607±0.163) μmol/L. After A549 cells was treated with celastrol, the cell number was significantly reduced, the cells were shrinked and round, the cell adhesion ability was weakened, the cell spacing becomeed larger, and the cell filopodia were reduced. Celastrol significantly inhibited the colony forming ability of A549 cells (P<0.05); celastrol significantly induced A549 apoptosis (P<0.05); celastrol significantly inhibited the invasion and migration ability of A549 cells (P<0.05); celastrol significantly inhibited COX-2 protein, MMP-2 protein, MMP-9 protein and p-p65/p65 in A549 cells expression(P<0.05); celastrol significantly inhibited the expression of COX-2 mRNA, MMP-2 mRNA and MMP-9 mRNA in A549 cells(P<0.05). Conclusions Celastrol can inhibit the proliferation, invasion and migration of NSCLC A549 cells, which may be related to the downregulation of COX-2, MMP-2, MMP-9 protein and mRNA expression through NF-κB signaling.
文章编号: 中图分类号: 文献标志码:A
基金项目:湖北省卫生健康科研基金(ZY2019M075)
Author Name | Affiliation |
LI Tao, YAN Hu, GONG Chang-zhi | Department of Intensive Care Unit, Huanggang Hospital of TCM, Huanggang, Hubei 438000, China |
Author Name | Affiliation |
LI Tao, YAN Hu, GONG Chang-zhi | Department of Intensive Care Unit, Huanggang Hospital of TCM, Huanggang, Hubei 438000, China |
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