摘要
目的:探讨高尔基体蛋白含酰基辅酶A结合结构域蛋白3(ACBD3)促进三阴性乳腺癌(TNBC)细胞增殖的作用及其机制。方法:通过qPCR和Westernblot检测乳腺癌细胞系中ACBD3的表达;使用沉默内源性ACBD3的TNBC细胞株,采用MTT实验、EdU染色、平板集落形成实验及裸鼠体内异种移植成瘤术检测ACBD3对细胞增殖能力的影响;通过基因集富集分析(GSEA)观察ACBD3与胞内信号通路的关系;Westernblot验证ACBD3对AKT-FOXOs信号通路相关蛋白的影响。结果:qPCR及Westernblot实验结果显示,ACBD3的mRNA及蛋白水平在乳腺癌细胞中均高于永生化正常乳腺上皮细胞MCF-10A,其中ACBD3在TNBC细胞中的表达水平尤其高;细胞功能学实验表明,ACBD3能够通过促进TNBC细胞G_(1)/S期的转变从而增强细胞的增殖能力;GSEA结果显示ACBD3的表达与AKT信号通路相关。Westernblot实验验证ACBD3能够通过增强AKT活性,抑制FOXOs活性而促进TNBC细胞增殖。结论:在TNBC细胞中ACBD3能够调控AKT-FOXOs信号通路而促进细胞增殖。
AIM:To investigate the effect of Golgi protein acyl-CoA binding domain containing protein 3(ACBD3)on the proliferation of triple-negative breast cancer(TNBC)cells and its possible mechanism.METHODS:The expression of ACBD3 in breast cancer cell lines was detected by qPCR and Western blot.MTT assay,EdU staining,colony formation assay and xenograft in nude mice were used to examine the effect of ACBD3 on the proliferation of ACBD3-silencing TNBC cells.The relationship between ACBD3 and AKT-FOXOs signaling pathway was analyzed by Western blot and gene set enrichment analysis(GSEA).RESULTS:Compared with normal breast epithelial cells(MCF-10A),the mRNA level of ACBD3 was highly expressed in breast cancer cells,especially in TNBC cell lines.In TNBC cells,ACBD3 enhanced cell proliferation by promoting the G_(1)/S phase transition.Furthermore,the results of GSEA showed that the ex-pression of ACBD3 was related to AKT-FOXOs signaling pathway.Western blot results confirmed that ACBD3 could promote the proliferation of TNBC cells by enhancing AKT activity and inhibiting FOXOs activity.CONCLUSION:The expression of ACBD3 in TNBC cells promotes the cell proliferation through AKT-FOXOs signaling pathway.
作者
郑迎春
汪兰
ZHENG Ying-chun;WANG Lan(School of Life Sciences and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou 510006,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2022年第8期1399-1406,共8页
Chinese Journal of Pathophysiology
基金
广东省自然科学基金资助项目(No.2017A030313549)。
