摘要
目的 探究中药泽漆对非小细胞肺癌(NSCLC)细胞PC-9和A549细胞的作用.方法 PC-9细胞和A549细胞体外培养,采用不同浓度泽漆作用于PC-9细胞(0、0.4、0.8、1.2 mg/mL)和A549细胞(0、0.8、1.2、1.6 mg/mL),CCK-8法和集落克隆实验检测泽漆对NSCLC细胞增殖的影响;流式细胞实验检测泽漆对NSCLC细胞凋亡的影响;划痕实验和Transwell实验检测泽漆对NSCLC细胞侵袭迁移的作用;Western blotting实验检测泽漆对NSCLC细胞凋亡和侵袭迁移相关蛋白Bax、Bcl-2、E-cadherin、vimentin、MMP2、MMP9表达的影响;将PC-9细胞注射到BALB/C裸鼠皮下,建立裸鼠皮下瘤动物模型,根据皮下瘤生长情况,小鼠被随机分为对照组:每日灌胃生理盐水;泽漆处理组:每日灌胃泽漆65 mg/mL,泽漆颗粒剂用生理盐水溶解;顺铂治疗组:每5 d腹腔注射顺铂4 mg/kg,每组6只小鼠,测量小鼠皮下肿瘤体积及质量变化,观察泽漆对荷瘤小鼠心、肝、脾、肺、肾的毒副作用以及治疗作用.结果 CCK-8和集落克隆实验结果显示泽漆呈浓度依赖性抑制PC-9和A549细胞增殖和存活(P<0.01);流式细胞实验结果显示泽漆诱导细胞凋亡(P<0.05);划痕实验和Transwell实验结果显示泽漆能够抑制NSCLC的侵袭迁移能力(P<0.05);Western blotting实验结果显示泽漆处理后上调E-Cadherin、Bax蛋白表达水平,下调Bcl-2、vimentin、MMP2、MMP9蛋白表达水平(P<0.05);动物实验结果显示:与对照组相比,泽漆能够抑制小鼠NSCLC皮下瘤的生长,并观察到对小鼠脏器没有产生明显毒性.结论 泽漆能够抑制NSCLC细胞增殖和侵袭迁移,诱导细胞发生凋亡.
Abstract
Objective To investigate the effect of Euphorbia helioscopia on biological behaviors of non-small cell lung cancer(NSCLC)cells.Methods NSCLC cell lines PC-9 and A549 treated with different concentrations of Euphorbia helioscopia preparations were examined for changes in proliferation,apoptosis,invasion and migration using CCK-8 assay,colony formation assay,flow cytometry,wound healing assay and Transwell assay.Western blotting was performed to detect the changes in protein expressions of Bax,Bcl-2,E-cadherin,vimentin,MMP2,and MMP9 in the treated cells.PC-9 cells were injected subcutaneously into BALB/C nude mice to establish a nude mouse subcutaneous tumor model.According to the growth of subcutaneous tumors,mice were randomly divided into control group:gavaged daily with saline;Euphorbia helioscopia-treated group:gavaged daily with Euphorbia helioscopia 65 mg/mL,and Euphorbia helioscopia granules were dissolved in saline;cisplatin-treated group:injected intraperitoneally with cisplatin 4 mg/kg every 5 days,6 mice per group.The subcutaneous tumor volume and mass changes of mice were measured,and the toxic effects of Euphorbia helioscopia on heart,liver,spleen,lung and kidney as well as the therapeutic effects of Euphorbia helioscopia were observed in the mice bearing tumor.Results Euphorbia helioscopia granules concentration-dependently inhibited the proliferation and survival of PC-9 and A549 cells,significantly promoted cell apoptosis,suppressed invasion and migration abilities of the cells,up-regulated the expression levels of E-cadherin and Bax,and down-regulated the expressions of Bcl-2,vimentin,MMP2,and MMP9.In the tumor-bearing mice,treatment with Euphorbia helioscopia significantly inhibited tumor growth without producing obvious toxicity in the vital organs.Conclusion Euphorbia helioscopia can inhibit proliferation,invasion,and migration and induces apoptosis of NSCLC cells in vitro.
基金项目
国家自然科学基金(82304640)
安徽省重点研究与开发计划项目(202104g01020017)
安徽省重点生化工程中心科研平台开放课题(2023SYKFD05)
安徽省自然科学基金(1908085QH373)
安徽高校自然科学研究项目(KJ2020A565)
安徽高校自然科学研究项目(KJ2021A0702)
安徽省优秀科研创新团队项目(2022AH010084)
安徽省大学生创新创业训练计划项目(S202310367111)
蚌埠医科大学研究生科研创新计划项目(Byycx22059)
蚌埠医科大学研究生科研创新计划项目(Byycx23053)
维普
万方数据