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硼酸对 A549 细胞和 A549 肿瘤干细胞中 DNA 双链断裂修复因子表达的影响:一项体外研究。

Boric Acid Affects the Expression of DNA Double-Strand Break Repair Factors in A549 Cells and A549 Cancer Stem Cells: An In Vitro Study.

机构信息

Cellular Therapy and Stem Cell Production, Application, and Research Center (ESTEM), Eskişehir Osmangazi University, Eskişehir, 26040, Turkey.

Department of Histology and Embryology, Faculty of Medicine, Eskisehir Osmangazi University, 26040, Eskişehir, Turkey.

出版信息

Biol Trace Elem Res. 2024 Nov;202(11):5017-5024. doi: 10.1007/s12011-024-04082-y. Epub 2024 Feb 17.

Abstract

DNA double-strand break (DSB) repair genes interact with tumor stemness- and resistance-associated processes in cancer stem cells (CSCs). Therefore, targeting DNA DSB genes in cancer treatment is important for the CSC phenotype. Although the anti-cancer effect of boric acid (BA) has been studied, its effect on DNA DSB is unclear. Moreover, no studies investigate BA's effects on DNA DSB of lung cancer stem cells (LC-SCs). To fill the gap, we aimed to assess the effects of BA on A549 cancer stem cells. CSCs were isolated from human non-small cell lung cancer cells (A549) and characterized by flow cytometry. Different concentrations of BA (at doses ranging from 1 to 100 mM) were applied to cancer stem cells. Cytotoxic activities were determined using the cell viability assay (MTT assay) at 24 and 48 h. Expression levels of DNA DSB genes that BRCA1, BRCA2, RAD51, KU70/80, ATM, and XRCC4 were evaluated by RT-qPCR. Additionally, immunofluorescence staining analysis was exploited for caspase-3 and E-cadherin. ATM expression increased significantly (p < 0.001). No significant change was observed in the expression of other genes. Moreover, BA up-regulated caspase-3 and E-cadherin expression. Consequently, we can say that BA affects DNA DSB and the apoptotic abilities of LC-SCs.

摘要

DNA 双链断裂 (DSB) 修复基因与肿瘤干细胞 (CSC) 中的肿瘤干性和耐药相关过程相互作用。因此,在癌症治疗中靶向 DNA DSB 基因对于 CSC 表型很重要。虽然硼酸 (BA) 的抗癌作用已经得到研究,但它对 DNA DSB 的影响尚不清楚。此外,尚无研究探讨 BA 对肺癌干细胞 (LC-SC) 的 DNA DSB 的影响。为了填补这一空白,我们旨在评估 BA 对 A549 癌症干细胞的影响。CSC 从人非小细胞肺癌细胞 (A549) 中分离出来,并通过流式细胞术进行表征。将不同浓度的 BA(剂量范围为 1 至 100 mM)应用于癌症干细胞。在 24 和 48 h 通过细胞活力测定 (MTT 测定) 测定细胞毒性活性。通过 RT-qPCR 评估 DNA DSB 基因 BRCA1、BRCA2、RAD51、KU70/80、ATM 和 XRCC4 的表达水平。此外,还利用免疫荧光染色分析评估了 caspase-3 和 E-钙粘蛋白。ATM 表达显著增加 (p < 0.001)。其他基因的表达没有观察到显著变化。此外,BA 上调了 caspase-3 和 E-钙粘蛋白的表达。因此,我们可以说 BA 影响 LC-SC 的 DNA DSB 和凋亡能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63be/11442501/32c991c95eb3/12011_2024_4082_Fig1_HTML.jpg

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