摘要
目的 探讨槐角黄酮对乳腺癌细胞增殖、迁移、侵袭和炎症因子分泌的影响.方法 将乳腺癌细胞BT549分为正常对照(NC)组、低剂量组、中剂量组、高剂量组、miR-NC组、miR-199a-3p组、高剂量+anti-miR-NC组、高剂量+anti-miR-199a-3p组.采用CCK-8、克隆形成实验评估细胞增殖;采用Transwell法评估细胞迁移和侵袭.采用实时荧光定量聚合酶链反应(RT-qPCR)检测miR-199a-3p表达.采用酶联免疫吸附测定(ELISA)法检测细胞培养液上清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)以及白细胞介素-1β(IL-1β)水平.采用蛋白免疫印迹(Western blot)检测p-p38MAPK和p-p65蛋白表达.结果 与NC组比较,低、中、高剂量组BT549细胞活力、克隆形成数、迁移数、侵袭数降低,miR-199a-3p表达升高,细胞培养液上清中TNF-α、IL-6以及IL-1β水平降低,差异均有统计学意义(P＜0.05).与miR-NC组比较,miR-199a-3p组BT549细胞活力、克隆形成数、迁移数、侵袭数降低,细胞培养液上清中TNF-α、IL-6以及IL-1β水平降低,差异均有统计学意义(P＜0.05).与高剂量+anti-miR-NC组比较,高剂量+anti-miR-199a-3p组BT549细胞活力、克隆形成数、迁移数、侵袭数升高,细胞培养液上清中TNF-α、IL-6以及IL-1β水平升高,差异均有统计学意义(P＜0.05).与NC组比较,高剂量组BT549细胞p-p38MAPK和p-p65蛋白表达降低,差异有统计学意义(P＜0.05).与高剂量+anti-miR-NC组比较,高剂量+anti-miR-199a-3p组BT549细胞p-p38MAPK和p-p65蛋白表达升高,差异有统计学意义(P＜0.05).结论 槐角黄酮可抑制乳腺癌细胞增殖、迁移、侵袭和炎症反应,其机制可能是通过上调miR-199a-3p表达、抑制p38MAPK/NF-κB通路实现的.
Abstract
Objective To investigate the effect of sophorae fructus flavonoids on cell prolifera-tion,migration,invasion and secretion of inflammatory factors in breast cancer.Methods The breast cancer cells BT549 were divided into normal control(NC)group,low-dose group,medium-dose group,high-dose group,miR-NC group,miR-199a-3p group,high-dose+anti-miR-NC group,and high-dose+anti-miR-199a-3p group.CCK-8 and clone formation assays were used to evaluate cell proliferation;Transwell method was performed to evaluate cell migration and invasion.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-199a-3p.EILSA method was used to detect the levels of tumor necrosis factor-α(TNF-α),in-terleukin-6(IL-6)and interleukin-1β(IL-1β)in the cell culture supernatant.Expression levels of p-p38MAPK and p-p65 proteins were detected by Western blot.Results Compared with the NC group,the cell viability,clone formation number,migration number and invasion number of BT549 cells in the low-,medium-and high-dose groups were significantly reduced,the expression of miR-199a-3p was significantly increased,and the levels of TNF-α,IL-6 and IL-1β in the cell culture supernatant were significantly reduced(P＜0.05).Compared with the miR-NC group,the cell viability,clone formation number,migration number and invasion number of BT549 cell in the miR-199a-3p group were significantly reduced,and the levels of TNF-α,IL-6 and IL-1β in the cell culture supernatant were significantly reduced as well(P＜0.05).Compared with the high-dose+anti-miR-NC group,the cell viability,clone formation number,migration number and invasion number of BT549 in the high-dose+anti-miR-199a-3p group were significantly increased,and the levels of TNF-α,IL-6 and IL-1β in the cell culture supernatant were significantly increased as well(P＜0.05).Compared with the NC group,the expression levels of p-p38MAPK and p-p65 proteins of BT549 cells in the high-dose group were significantly reduced(P＜0.05).Compared with the high-dose+anti-miR-NC group,the expression levels of p-p38MAPK and p-p65 proteins of BT549 cells in high-dose+anti-miR-199a-3p group were significantly increased(P＜0.05).Conclusion Sophorae fructus flavonoids can inhibit cell proliferation,migration,invasion and inflammation reaction in breast cancer,and its mechanism may be achieved by up-regulating the expression of miR-199a-3p and in-hibiting p38MAPK/NF-κB pathway.
维普
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