车叶草苷调节HMGB1-RAGE信号通路对乳腺癌细胞恶性生物学行为的影响  

Effect of asperuloside on the malignant biological behavior of breast cancer cells by regulating HMGB1-RAGE signal pathway

作　　者：杨森焱 张志东 彭文 陈吉柏 薛治乾 YANG Senyan;ZHANG Zhidong;PENG Wen;CHEN Jibai;XUE Zhiqian(Department of Thoracic Surgery,Danzhou People’s Hospital,Danzhou,Hainan 571700,China;Department of Pathology,Danzhou People’s Hospital,Danzhou,Hainan 571700,China)

机构地区：[1]儋州市人民医院胸外乳甲外科,海南儋州571700 [2]儋州市人民医院病理科,海南儋州571700

出　　处：《中国优生与遗传杂志》2024年第12期2494-2500,共7页Chinese Journal of Birth Health & Heredity

摘　　要：目的探究车叶草苷(ASP)是否通过调节高迁移率组蛋白1(HMGB1)/晚期糖基化终产物受体(RAGE)信号通路影响乳腺癌细胞的恶性生物学行为。方法使用CCK8检测不同浓度ASP处理HCC1937细胞24 h的增殖情况并计算IC50,将HCC1937细胞分为5组:Control组(HCC1937细胞)、L-ASP组(3 mmol/L ASP处理HCC1937细胞)、H-ASP组(4 mmol/L ASP处理HCC1937细胞)、ASP-NC组(pcDNA3.1-NC+4 mmol/L ASP)、ASP-HMGB1组(pcDNA-HMGB1+4mmol/L ASP)。利用细胞克隆实验检测细胞增殖;利用细胞划痕实验、Transwell实验和Hoechst33342染色实验检测HCC1937细胞的迁移、侵袭和凋亡;利用RT-qPCR实验检测HMGB1、RAGE基因表达;利用Western blotting实验检测HMGB1、肿瘤坏死因子α(TNF-α)、白细胞介素-4(IL-4)、RAGB、白细胞介素-1β(IL-1β)蛋白表达;利用ELISA实验检测细胞上清中相关促炎因子IL-1β、抗炎因子IL-4和TNF-α的含量。结果与Control组比,L-ASP组、H-ASP组细胞增殖、迁移、侵袭、蛋白(HMGB1、IL-1β、RAGB、TNF-α)表达、基因(HMGB1、RAGB)表达及细胞上清TNF-α、IL-1β含量降低(P<0.05),细胞凋亡和IL-4蛋白表达及细胞上清IL-4含量增加(P<0.05);与ASP-NC组相比,ASP-HMGB1组细胞增殖、迁移、侵袭、上述蛋白表达及细胞上清TNF-α、IL-1β含量升高(P<0.05),细胞凋亡、IL-4蛋白表达及细胞上清IL-4含量减少(P<0.05)。结论ASP可以抑制乳腺癌细胞恶性生物学行为,其机制可能是通过抑制HMGB1/RAGE信号通路实现的。Objective To investigate whether asperuloside(ASP)affects the malignant biological behavior of breast cancer cells by regulating the high mobility group protein 1(HMGB1)/receptor of advanced glycation endproducts(RAGE)signaling pathway.Methods CCK8 was applied to detect the proliferation of HCC1937 cells treated with different concentrations of ASP for 24 h and IC50 was calculated.HCC1937 cells were divided into five groups:Control group(HCC1937 cells),L-ASP group(3 mmol/L ASP treated HCC1937 cells),H-ASP group(4 mmol/L ASP treated HCC1937 cells),ASP-NC group(pcDNA3.1-NC+4 mmol/L ASP),and ASP-HMGB1 group(pcDNA-HMGB1+4 mmol/L ASP).Cell cloning assay was applied to detect proliferation.Cell scratch assay,Transwell assay,and Hoechst33342 staining assay were applied to detect the ability of migration,invasion,and apoptosis of HCC1937 cells.RT-qPCR experiments were applied to detect the expression of HMGB1 and RAGE genes.Western blotting experiments were applied to detect the expression of HMGB1,tumor necrosis factor-α(TNF-α),interleukin 4(IL-4),RAGB,and interleukin 1β(IL-1β)proteins.ELISA assay was applied to detect the levels of pro-inflammatory factor IL-1β,anti-inflammatory factor IL-4,and TNF-αin the cell supernatant.Results Compared with the control group,the proliferation,migration,invasion,expression of proteins(HMGB1,IL-1β,RAGB,TNF-α),expression of genes(HMGB1,RAGB),and contents of TNF-αand IL-1βin the cell supernatant were decreased in the L-ASP and H-ASP groups(P<0.05),while apoptosis,expression of IL-4 protein,and content of IL-4 in the cell supernatant were increased(P<0.05).Compared with the ASP-NC group,the cell proliferation,migration,invasion,expression of the aforementioned proteins,and contents of TNF-αand IL-1βin the cell supernatant were increased in ASP-HMGB1 group(P<0.05),while apoptosis,expression of IL-4 protein,and content of IL-4 in the cell supernatant were decreased(P<0.05).Conclusion ASP can inhibit the malignant biological behavior of breast cancer cells,and may be achieved b

关 键 词：车叶草苷 乳腺癌 高迁移率组蛋白1/晚期糖基化终产物受体 恶性生物学行为 

分 类 号：R737.9[医药卫生—肿瘤]