摘要
目的 探讨双氢青蒿素(DHA)对人口腔鳞癌细胞增殖能力的影响及其作用机制.方法 用不同浓度的双氢青蒿素干预CAL27细胞,采用CCK-8法检测其细胞增殖活力,集落形成实验检测细胞克隆形成能力;基于网络药理学和生物信息学筛选DHA抑制口腔癌生物学行为的潜在靶点;不同浓度的DHA干预CAL27细胞后,Western blot检测增殖相关蛋白PCNA和自噬相关蛋白Beclin-1、LC3的表达;联合自噬阻断剂3-甲基腺嘌呤和自噬诱导剂雷帕霉素与双氢青蒿素共处理细胞后,检测细胞增殖活力、克隆形成能力和增殖及自噬相关蛋白的表达.结果 双氢青蒿素显著降低了CAL27细胞的增殖活力及克隆形成能力,且呈现浓度依赖性,PCNA的表达量也显著下降.网络药理学结合生物信息学发现DHA抑制口腔癌的靶点涉及自噬相关通路.DHA干预可升高细胞内自噬相关蛋白Beclin-1、LC3的表达,DHA联合自噬阻断剂共处理CAL27后,细胞的增殖活力及克隆形成能力降低,PCNA的表达升高、Beclin-1、LC3的表达降低.结论 双氢青蒿素可在体外抑制口腔鳞癌细胞的增殖能力,其作用机制可能与诱导细胞自噬相关.
Abstract
Objective To investigate the effect of dihydroartemisinin(DHC)on the proliferation capacity of human oral squamous carcinoma cells and its mechanism of action.Methods The viability and colony formation ability of CAL27 cells treated with different concentrations of dihydroartemisinin was measured by CCK-8 and colony formation assay.The expression of proteins related to proliferation and autophagy was determined by Western blot.Potential targets for DHA inhibition of the biological behavior of oral cancer were screened based on network pharmacology and bioinformatics.Measurement was conducted after the cells were cotreated with autophagy blocker 3-methyladenine and autophagy inducers rapamycin and dihydroartemisinin.Results Dihydroartemisinin significantly reduced the proliferation viability and clone formation ability of CAL27 cells in a concentration-dependent manner.The PCNA expression level also decreased substantially.DHA suppressed oral cancer targets involving autophagy-related pathways.DHA intervention increased the expression of intracellular autophagy-related proteins Beclin-1 and LC3.After co-treatment of DHA combined with autophagy blocker,the proliferation viability and clone formation ability of CAL27 cells decreased.The expression of PCNA increased,and the expression of Beclin-1 and LC3 decreased.Conclusion Dihydroartemisinin could inhibit the proliferative capacity of oral squamous carcinoma cells in vitro,and its effect may be correlated with the induction of autophagy.
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