摘要
目的探讨“聊红椿”提取物对非小细胞肺癌的抑制效果,研究其作用机制。方法通过有机溶剂萃取法,提取“聊红椿”中有效成分;将其作用于人肺癌细胞系A549,MTT法检测A549细胞增殖情况;流式细胞仪考察A549细胞凋亡情况;进一步通过荧光定量PCR和Western blot考察细胞凋亡因子Caspase-3,Bax,Bcl-2,cleaved PARP的mRNA和蛋白水平表达情况;构建肺癌裸小鼠皮下移植瘤模型,将低、中、高剂量“聊红椿”提取物分别腹腔注射干预治疗,作用一定时间后,测量小鼠肿瘤生长情况。结果HPLC分析发现,与正常臭椿相比,“聊红椿”提取物中有高含量有效成分臭椿酮;MTT实验结果发现,“聊红椿”提取物能够明显抑制A549细胞增殖;流式细胞仪检测结果发现,“聊红椿”提取物能够诱导A549细胞发生凋亡;荧光定量PCR和Western blot实验结果发现,“聊红椿”提取物能够使得细胞凋亡因子Caspase-3,Bax和cleaved PARP含量显著下降,抗凋亡因子Bcl-2含量显著增加;“聊红椿”提取物作用于肺癌裸小鼠皮下移植瘤模型后,肿瘤质量和直径明显下降,且与处理浓度呈反比。结论“聊红椿”提取物能够通过影响相关细胞凋亡因子的表达,显著抑制非小细胞肺癌细胞增殖,促进细胞凋亡。
Objective To explore the inhibitory effect of‘Liaohongchun’extract on non-small cell lung cancer,and to study its mechanism of action.Methods The effective ingredients in‘Liaohongchun’were extracted by organic solvent extraction;It was applied to human lung cancer cell line A549,MTT method was used to detect the proliferation of A549 cells;Flow cytometry was used to investigate the apoptosis of A549 cells;The mRNA and protein levels of apoptosis factors Caspase-3,Bax,Bcl-2 and cleaved PARP were further investigated by fluorescence quantitative PCR and Western blot Expression status;Constructed a subcutaneous xenograft tumor model of lung cancer in nude mice,and intraperitoneally inject low,medium,and high doses of‘Liaohongchun’extract for intervention therapy.After a certain period of action,the tumor growth of mice was measured.Results HPLC analysis found that,compared with normal Ailanthus altissima,there is a high content of active ingredient Ailanthone in the‘Liaohongchun’extract;MTT experiment found that the‘Liaohongchun’extract can significantly inhibited the proliferation of A549 cells;The results of flow cytometry found that‘Liaohongchun’extract can induce apoptosis of A549 cells;The results of fluorescence quantitative PCR and Western blot experiments found that‘Liaohongchun’extract can significantly reduce the content of apoptosis factors Caspase-3,Bax and cleaved PARP,the content of apoptosis factor Bcl-2 increased significantly;After the‘Liaohongchun’extract was applied to the subcutaneous transplanted tumor model of lung cancer in nude mice,the tumor mass and diameter decreased significantly,and it was inversely proportional to the treatment concentration.Conclusion‘Liaohongchun’extract can significantly inhibit the proliferation of non-small cell lung cancer cells and promote apoptosis by affecting the expression of related apoptosis factors.
作者
张泽鑫
李攀
马晓瑞
谢芳
魏洪帅
朱艳娟
冀芦沙
ZHANG Zexin;LI Pan;MA Xiaorui;XIE Fang;WEI Hongshuai;ZHU Yanjuan;JI Lusha(School of Pharmacy,Liaocheng University,Liaocheng 250059,China;Yinchuan Center for Disease Control and Prevention,Yinchuan 750000,China)
出处
《聊城大学学报(自然科学版)》
2021年第1期104-110,共7页
Journal of Liaocheng University:Natural Science Edition
基金
国家重点研发项目(SQ2020YFF0422322)
山东省自然科学基金项目(ZR2019PC041)资助。