盐酸青藤碱通过抑制bFGF介导的乳腺癌血管新生发挥抑瘤作用

Sinomenine hydrochloride suppresses breast cancer growth by inhibiting bFGF-induced angiogenesis

目的初步探索盐酸青藤碱对乳腺癌血管新生的作用及其潜在机制。方法不同浓度盐酸青藤碱干预4T1乳腺癌原位移植瘤模型,观察其对肿瘤生长的影响;对瘤组织中的内皮细胞标记CD31进行免疫组化染色,分析新生血管的面积变化;用ELISA法检测肿瘤组织间液和肿瘤细胞裂解液中碱性成纤维细胞生长因子(bFGF)浓度,分析bFGF的分泌和生成变化;RT-qPCR检测肿瘤细胞内bFGF的mRNA变化;采用在线生存分析对比bFGF不同表达水平与乳腺癌患者预后的关系。结果与对照组比较,100 mg/kg盐酸青藤碱组可抑制4T1原位移植瘤的生长(P<0.05),而体外研究显示盐酸青藤碱对4T1细胞系未见显著细胞毒作用;100 mg/kg盐酸青藤碱组移植瘤内新生血管面积较对照组减少(P<0.01),且盐酸青藤碱组肿瘤组织间液中及肿瘤细胞内bFGF浓度较对照组下降(P<0.05),盐酸青藤碱组肿瘤细胞内bFGF的mRNA表达较对照组下降(P<0.05);Kaplan-Meier Plotter在线生存分析提示,乳腺癌患者肿瘤组织中bFGF的高表达与较差的总生存、无复发生存和无远处转移生存相关(P<0.01)。结论盐酸青藤碱可通过抑制bFGF介导的血管新生在乳腺癌中发挥抑瘤作用,这丰富了该中药的药理学作用。

Objective To investigate the effect of sinomenine hydrochloride(SH)on angiogenesis and the underlying mechanism in breast cancer.Methods The 4T1 orthotopic tumor model of breast cancer was utilized,and mice were treated with different dosage of SH to investigate the effect of SH on tumor growth.IHC staining of CD31 was used to evaluate angiogenesis within tumors under different treatment.ELISA was performed to measure the bFGF level within tumor extracellular fluid(TEF)and tumor cells to analyze the effect of SH on bFGF secretion and production.RTqPCR was utilized to evaluate the effect of SH on the mRNA expression of bFGF in tumor cells.Kaplan-Meier Plotter was analyzed online to investigate the relationship between bFGF expression and the survival of patients with breast cancer.Results SH at 100 mg/kg could inhibit 4T1 orthotopic tumor growth compared with saline(P<0.05),but SH showed little cytotoxicity in vitro under the tested concentrations.SH at 100 mg/kg suppressed the vessel area compared with saline(P<0.01),and the concentration of angiogenic factor bFGF in TEF and tumor cells was decreased by SH treatment(P<0.05),while the mRNA expression of bFGF in tumor cells was also downregulated by SH treatment(P<0.05).Kaplan-Meier Plotter online analysis showed that elevated mRNA expression of bFGF in the primary tumor was associated with poorer OS,RFS and DMFS in patients with breast cancer(P<0.01).Conclusion SH inhibits breast cancer growth by suppressing bFGF-induced angiogenesis,which enriches the pharmacological effects of this traditional Chinese medicine.