Feng Xiuyan, Han Han, Guo Yarui, Feng Xue, Guo Shanchun, Zhou Weiqiang
Medical Administration Division, The Second Affiliated Hospital of Shenyang Medical College, Shenyang City, China.
Department of Biochemistry and Molecular Biology, Shenyang Medical College, Shenyang City, China.
Front Oncol. 2022 Apr 4;12:874343. doi: 10.3389/fonc.2022.874343. eCollection 2022.
Breast cancer is one of the leading threats to the health of women. It has the highest incidence and mortality in women worldwide. Although progress has been made in the development and application of anti-breast cancer drugs such as Chidamide and others, the occurrence of drug resistance limits the effective application of chemotherapies. The purpose of this study is to explore the role of LncRNA in the pharmacological effect of Chidamide in breast cancer therapy. The human breast cancer MCF-7 or MDA-MB-231 cells were used as the research cell models. The RNA library screening and high-throughput sequencing comparative analysis was conducted. The binding of LncRNA and its downstream target genes in RNA and protein levels was tested. The results showed that the expression of LncRNA ENST869 in cells treated with Chidamide increased significantly, as demonstrated by real-time PCR and cell viability assay. RNAplex analysis showed that LncRNA ENST869 and Nestin mRNA may interact. RNA interference and Western blot analysis indicated that LncRNA ENST869 could target and regulate the expression of Nestin. Luciferase assay and RNA-protein pulldown showed that LncRNA ENST869 affected Nestin transcription. There might be a highly active binding region of LncRNA ENST869 in regulating Nestin transcriptional activity within the site of 250 bp upstream of the transcription starting point of Nestin. In addition, LncRNA ENST869 did not directly interact with Nestin protein to affect its activity. In conclusion, our results demonstrated that LncRNA ENST869 could affect the function of Nestin in breast cancer cells treated with Chidamide. Nestin is a key player in influencing the pharmacological activity of Chidamide and an essential factor in drug resistance of breast cancer cells.
乳腺癌是女性健康的主要威胁之一。它在全球女性中发病率和死亡率最高。尽管在诸如西达本胺等抗乳腺癌药物的研发和应用方面取得了进展，但耐药性的出现限制了化疗的有效应用。本研究的目的是探讨长链非编码RNA（LncRNA）在西达本胺治疗乳腺癌的药理作用中的作用。将人乳腺癌MCF-7或MDA-MB-231细胞用作研究细胞模型。进行了RNA文库筛选和高通量测序比较分析。检测了LncRNA及其下游靶基因在RNA和蛋白质水平的结合情况。结果表明，经西达本胺处理的细胞中LncRNA ENST869的表达显著增加，实时定量聚合酶链反应（PCR）和细胞活力测定证明了这一点。RNAplex分析表明LncRNA ENST869和巢蛋白（Nestin）mRNA可能相互作用。RNA干扰和蛋白质免疫印迹分析表明LncRNA ENST869可以靶向并调节Nestin的表达。荧光素酶报告基因检测和RNA-蛋白质下拉实验表明LncRNA ENST869影响Nestin转录。在Nestin转录起始点上游250 bp位点内，LncRNA ENST869在调节Nestin转录活性方面可能存在一个高活性结合区域。此外，LncRNA ENST869不直接与Nestin蛋白相互作用以影响其活性。总之，我们的结果表明LncRNA ENST869可以影响西达本胺处理的乳腺癌细胞中Nestin的功能。Nestin是影响西达本胺药理活性的关键因素，也是乳腺癌细胞耐药的重要因素。
