小檗碱增强非小细胞肺癌放疗敏感性和下调STAT3信号通路分子水平

Berberine enhances radiosensitivity of non-small cell lung cancer and downregulates levels of molecules involved in STAT3 signaling pathway

目的探讨小檗碱对非小细胞肺癌放疗敏感性的影响及其作用机制。方法分别用放疗、小檗碱处理、小檗碱联合放疗非小细胞肺癌Calu-1细胞后,通过MTT法检测细胞活性,流式细胞术检测细胞凋亡。构建Calu-1细胞异位移植瘤裸鼠模型,分别采用放疗、小檗碱处理和小檗碱联合放疗处理异位移植瘤裸鼠后,测量移植瘤的体积和质量;通过HE染色观察移植瘤模型裸鼠的肺、肝、脾和肾组织的病理学结构,TUNEL染色检测瘤体组织的凋亡,免疫组织化学法检测瘤体组织中p-STAT3、HIF-1α和cyclin D1水平。结果在体外,与放疗组比较,小檗碱联合放疗组Calu-1细胞活性明显降低,细胞凋亡明显增加。在Calu-1细胞异位移植瘤模型裸鼠,与放疗组比较,小檗碱联合放疗组瘤体的体积、质量以及瘤体组织中p-STAT3、HIF-1α和cyclin D1蛋白表达均明显降低;HE染色显示,3种处理方式均未对肺、肝、脾和肾组织产生毒性反应。结论小檗碱在体内和体外均增强Calu-1细胞的放疗敏感性,其机制可能与降低Calu-1细胞的p-STAT3及其下游信号分子HIF-1α和cyclin D1水平相关。

Objective To investigate the effect of berberine on radiosensitivity of non-small cell lung cancer and its mechanism.Methods Calu-1 cells of non-small cell lung cancer were treated with radiotherapy,berberine treatment,and radiotherapy combined with berberine treatment,respectively.After the treatments,cell viability was detected by MTT assay,and cell apoptosis was detected by flow cytometry.A nude mouse model of Calu-1 cells xenograft tumor was established and treated with radiotherapy,berberine treatment and berberine combined with radiotherapy,respectively,then the volume and weight of xenografted tumors were measured.The pathological structure of lung,liver,spleen and kidney in nude mice was observed by HE staining.Cellular apoptosis of tumor tissues was detected by TUNEL staining,and the expression of p-STAT3,HIF-1αand cyclin D1 in tumor tissues were detected by immunohistochemistry.Results In vitro,cell viability of Calu-1 cells was significantly decreased and cellular apoptosis was significantly increased in berberine combined with radiotherapy group compared to the radiotherapy group.In vivo,the volume and weight of the xenografted tumors and the expression of p-STAT3,HIF-1αand cyclin D1 in tumor tissues were significantly decreased in berberine combined with radiotherapy group compared to the radiotherapy group.Meanwhile,the results of HE staining showed that none of these three treatment methods had toxic effects on lung,liver,spleen and kidney tissues.Conclusion Berberine enhances radiotherapy sensitivity of Calu-1 cells in vivo and in vitro,and the mechanism may be related to the decreased expression of p-STAT3 and its downstream signaling molecules HIF-1αand cyclin D1 in Calu-1 cells.