整合转录组和体外分析揭示过硼酸钠对肝癌细胞的抗增殖作用。

Integrated transcriptome and in vitro analysis revealed anti-proliferative effect of sodium perborate on hepatocellular carcinoma cells.

机构信息

Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, Kayisdagi Cad., Atasehir, Istanbul 34755, Turkey.

Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, Kayisdagi Cad., Atasehir, Istanbul 34755, Turkey; Department of Molecular Biology and Genetics, Faculty of Engineering and Natural Sciences, Istanbul Sabahattin Zaim University, Halkali Cad. 281, Kucukcekmece, Istanbul 34303, Turkey.

出版信息

J Trace Elem Med Biol. 2022 Sep;73:127011. doi: 10.1016/j.jtemb.2022.127011. Epub 2022 Jun 3.

DOI:10.1016/j.jtemb.2022.127011

PMID:35716648

Abstract

BACKGROUND

Hepatocelular carcinoma is one of the leading cancer types with no effective cure as poor prognosis is still a challenging aspect. Thus, alternative therapeutics are necessary to control hepatocelular carcinoma. Boron derivatives such as boric acid (BA), sodium perborate tetrahydrate (SPT) and sodium pentaborate pentahydrate (NaB) have been discovered to have anti-cancer effect. This study investigated the anti-proliferative effects of SPT against hepatocelular carcinoma (HCC) using in vitro and transcriptome approaches.

METHODS

Cytotoxic level of SPT on cell survival were detected using MTS assay. The apoptotic cell death and cell cycle arrest was determined using Annexin V/PI and cell cycle assay, respectively. Transcriptome analysis was performed using RNA-seq, followed by functional and KEGG pathway enrichment analysis. qPCR was used to validate the different genes.

RESULTS

SPT treated HepG2 and Hep3B cells induced cytotoxicity having IC values of 1.13 mM and 0.91 mM, respectively. SPT caused mitotic arrest in G/G phase at 48 h and subsequent apoptotic cell death. RNA-seq revealed a total number of 822 and 1075 differentially expressed genes (DEGs) which after SPT treatment in HepG2 and Hep3B cells, respectively. Functional and KEGG pathway enrichment results suggested that there are several genes involved to induce apoptosis related pathways. The DEGs in p53 signaling pathway may have closely relationships to the cells apoptosis caused by SPT treatment. qPCR results validated dynamic changes in p53 signaling pathway, DNA replication and cell cycle related genes, such as CDKN1A, SERPINE1, PMAIP1, MCM3, MCM5 and MCM6.

CONCLUSION

In vitro experiments and RNA-seq analysis show anti-proliferative and apoptotic effect of SPT in HCC cells. Further studies might help in understanding the molecular mechanisms of SPT.

摘要

背景

肝细胞癌是主要癌症类型之一，由于预后较差，目前尚无有效治疗方法。因此，需要替代疗法来控制肝细胞癌。硼酸（BA）、过硼酸钠四水合物（SPT）和五硼酸钠十水合物（NaB）等硼衍生物已被发现具有抗癌作用。本研究采用体外和转录组学方法研究 SPT 对肝癌（HCC）的抗增殖作用。

方法

用 MTS 法检测 SPT 对细胞存活的细胞毒性。用 Annexin V/PI 和细胞周期分析分别测定凋亡细胞死亡和细胞周期阻滞。用 RNA-seq 进行转录组分析，然后进行功能和 KEGG 途径富集分析。用 qPCR 验证不同基因。

结果

SPT 处理 HepG2 和 Hep3B 细胞诱导细胞毒性，IC 值分别为 1.13 mM 和 0.91 mM。SPT 在 48 小时引起有丝分裂阻滞在 G/G 期，随后发生凋亡细胞死亡。RNA-seq 显示 SPT 处理后 HepG2 和 Hep3B 细胞分别有 822 和 1075 个差异表达基因（DEGs）。功能和 KEGG 途径富集结果表明，有几个基因参与诱导凋亡相关途径。p53 信号通路中的 DEGs 可能与 SPT 处理引起的细胞凋亡密切相关。qPCR 结果验证了 p53 信号通路、DNA 复制和细胞周期相关基因（如 CDKN1A、SERPINE1、PMAIP1、MCM3、MCM5 和 MCM6）的动态变化。