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Abstract：
BACKGROUND: Because of the progress on the diagnosis and treatment for patients with breast cancer (BC), the overall survival of the patients has been improved. However, a number of BC patients cannot benefit from the existing therapeutic strategies as the essential molecular events triggering the development of BC are not well understood. Previous studies have shown that abnormal expression of zinc finger proteins is involved in the development of various malignancies, whereas it remains largely unclear on their significance during the progression of BC. In this study, we aimed to explore the clinical relevance, cellular function and underlying mechanisms of zinc finger protein 468 (ZNF468) in BC.
METHODS: The clinical relevance of ZNF468 and TFAM was analyzed based on TCGA database. Overexpression or knockdown of ZNF468 and TFAM were performed by transfecting the cells with overexpression plasmids and siRNAs, respectively. Overexpression and knockdown efficacy was checked by immunoblotting. CCK-8, colony formation, transwell and apoptosis experiments were conducted to check the cellular function of ZNF468 and TFAM. The content of mtDNA was measured by the indicated assay kit. The effects of cisplatin on BC cells were detected by CCK-8 and colony formation assays. The regulation of ZNF468 on TFAM was analyzed by RT-qPCR, immunoblotting, dual luciferase activity and ChIP-qPCR assays.
RESULTS: ZNF468 was overexpressed in BC patients and inversely correlated with their prognosis. Based on overexpression and knockdown assays, we found that ectopic expression of ZNF468 was essential for the proliferation, growth and migration of BC cells. The expression of ZNF468 also negatively regulated the sensitivity of BC cells to the treatment of cisplatin. Mechanistically, ZNF468 potentiated the transcription activity of TFAM gene via direct binding on its promoter. Lastly, we demonstrated that ZNF468 up-regulation of TFAM was important for the growth, migration and cisplatin resistance in BC cells.
CONCLUSIONS: Our study indicates that ZNF468 promotes BC cell growth and migration via transcriptional activation of TFAM. ZNF468/TFAM axis can serve as the diagnostic and therapeutic target, as well as the predictor of cisplatin effectiveness in BC patients.
METHODS: The clinical relevance of ZNF468 and TFAM was analyzed based on TCGA database. Overexpression or knockdown of ZNF468 and TFAM were performed by transfecting the cells with overexpression plasmids and siRNAs, respectively. Overexpression and knockdown efficacy was checked by immunoblotting. CCK-8, colony formation, transwell and apoptosis experiments were conducted to check the cellular function of ZNF468 and TFAM. The content of mtDNA was measured by the indicated assay kit. The effects of cisplatin on BC cells were detected by CCK-8 and colony formation assays. The regulation of ZNF468 on TFAM was analyzed by RT-qPCR, immunoblotting, dual luciferase activity and ChIP-qPCR assays.
RESULTS: ZNF468 was overexpressed in BC patients and inversely correlated with their prognosis. Based on overexpression and knockdown assays, we found that ectopic expression of ZNF468 was essential for the proliferation, growth and migration of BC cells. The expression of ZNF468 also negatively regulated the sensitivity of BC cells to the treatment of cisplatin. Mechanistically, ZNF468 potentiated the transcription activity of TFAM gene via direct binding on its promoter. Lastly, we demonstrated that ZNF468 up-regulation of TFAM was important for the growth, migration and cisplatin resistance in BC cells.
CONCLUSIONS: Our study indicates that ZNF468 promotes BC cell growth and migration via transcriptional activation of TFAM. ZNF468/TFAM axis can serve as the diagnostic and therapeutic target, as well as the predictor of cisplatin effectiveness in BC patients.
摘要：
背景：由于乳腺癌（BC）患者的诊断和治疗进展，患者的总体生存率得到改善.然而,一些BC患者不能从现有的治疗策略中获益,因为对引发BC发展的基本分子事件了解不多.以往的研究表明，锌指蛋白的异常表达参与各种恶性肿瘤的发展，而它们在BC进展过程中的意义仍不清楚。在这项研究中,我们旨在探索临床相关性，锌指蛋白468（ZNF468）在BC中的细胞功能和潜在机制。
方法：基于TCGA数据库分析ZNF468和TFAM的临床相关性。通过用过表达质粒和siRNA转染细胞进行ZNF468和TFAM的过表达或敲减，分别。通过免疫印迹检查过表达和敲低功效。CCK-8,集落形成,进行了transwell和凋亡实验以检查ZNF468和TFAM的细胞功能。通过指定的测定试剂盒测量mtDNA的含量。通过CCK-8和集落形成试验检测顺铂对BC细胞的影响。通过RT-qPCR分析ZNF468对TFAM的调控,免疫印迹,双荧光素酶活性和ChIP-qPCR测定。
结果：ZNF468在BC患者中过度表达，与他们的预后呈负相关。基于过表达和敲低测定，我们发现ZNF468的异位表达对于增殖至关重要，BC细胞的生长和迁移。ZNF468的表达也负调控了BC细胞对顺铂医治的敏理性。机械上,ZNF468通过直接结合其启动子来增强TFAM基因的转录活性。最后,我们证明了ZNF468上调TFAM对于生长是重要的，BC细胞的迁移和顺铂抗性。
结论：我们的研究表明ZNF468通过TFAM的转录激活促进BC细胞生长和迁移。ZNF468/TFAM轴可作为诊断和治疗靶点,以及顺铂在BC患者中的疗效预测因子。
方法：基于TCGA数据库分析ZNF468和TFAM的临床相关性。通过用过表达质粒和siRNA转染细胞进行ZNF468和TFAM的过表达或敲减，分别。通过免疫印迹检查过表达和敲低功效。CCK-8,集落形成,进行了transwell和凋亡实验以检查ZNF468和TFAM的细胞功能。通过指定的测定试剂盒测量mtDNA的含量。通过CCK-8和集落形成试验检测顺铂对BC细胞的影响。通过RT-qPCR分析ZNF468对TFAM的调控,免疫印迹,双荧光素酶活性和ChIP-qPCR测定。
结果：ZNF468在BC患者中过度表达，与他们的预后呈负相关。基于过表达和敲低测定，我们发现ZNF468的异位表达对于增殖至关重要，BC细胞的生长和迁移。ZNF468的表达也负调控了BC细胞对顺铂医治的敏理性。机械上,ZNF468通过直接结合其启动子来增强TFAM基因的转录活性。最后,我们证明了ZNF468上调TFAM对于生长是重要的，BC细胞的迁移和顺铂抗性。
结论：我们的研究表明ZNF468通过TFAM的转录激活促进BC细胞生长和迁移。ZNF468/TFAM轴可作为诊断和治疗靶点,以及顺铂在BC患者中的疗效预测因子。
