Li Feng, Zhi Yuanyuan, Wang Yinghui, Hussain Shaik Althaf, Alrubie Turki Mayudh, Yang Ping
Department of Obstetrics and Gynecology, Xi'an Third Hospital, Xi'an, China.
Department of Gynecology, Shandong Provincial Maternal and Child Health Care Hospital, Jinan, China.
Front Oncol. 2025 Jun 12;15:1567493. doi: 10.3389/fonc.2025.1567493. eCollection 2025.
Investigated within the endometrial carcinoma (EC) context, Snail-1 emerges as a pivotal transcription factor governing invasion and metastasis by orchestrating epithelial-to-mesenchymal transition (EMT). Employing small interfering RNA (siRNA) to silence Snail-1 expression in the HEC-1A cell line, this study explored the repercussions on the expression of genes implicated in metastasis, cellular cytotoxicity, apoptosis, and migration.
HEC-1A cells were transfected with Snail-1-specific siRNA. Quantitative Real-time PCR was utilized to quantify the mRNA levels of , Matrix metalloproteinase-9 , , and . Western blot analysis was also performed to ascertain alterations in Snail-1, MMP-9, Vimentin, E-cadherin, and Notch1 protein levels. Cytotoxicity of transfected cells was assessed via the MTT assay, while flow cytometry was employed to measure apoptosis. Migration was evaluated using a wound healing assay.
Transfection with 60 pmol/mL of Snail-1-specific siRNA significantly reduced Snail-1 expression at both the mRNA and protein levels. This was accompanied by decreased MMP-9, Vimentin, and Notch1 expression and increased E-cadherin expression, all confirmed at both transcript and protein levels. Furthermore, gene expression analysis revealed a downregulation of and mRNA levels and an upregulation of miR-34a. Moreover, transfection correlated with increased apoptosis and decreased migration of treated HEC-1A cells.
The study emphasizes the significant influence of Snail-1 on EMT in EC cells, thereby impacting apoptosis and metastasis. Targeted silencing of Snail-1 through specific siRNA emerges as a promising therapeutic approach in treating EC.
在子宫内膜癌(EC)背景下进行研究时,Snail-1作为一种关键转录因子出现,它通过协调上皮-间质转化(EMT)来控制侵袭和转移。本研究利用小干扰RNA(siRNA)使HEC-1A细胞系中的Snail-1表达沉默,探讨其对与转移、细胞毒性、凋亡和迁移相关基因表达的影响。
用Snail-1特异性siRNA转染HEC-1A细胞。采用定量实时PCR定量基质金属蛋白酶-9、、和的mRNA水平。还进行了蛋白质印迹分析,以确定Snail-1、MMP-9、波形蛋白、E-钙黏蛋白和Notch1蛋白水平的变化。通过MTT试验评估转染细胞的细胞毒性,同时采用流式细胞术测量凋亡。使用伤口愈合试验评估迁移情况。
用60 pmol/mL的Snail-1特异性siRNA转染显著降低了Snail-1在mRNA和蛋白质水平的表达。这伴随着MMP-9、波形蛋白和Notch1表达的降低以及E-钙黏蛋白表达的增加,所有这些在转录本和蛋白质水平均得到证实。此外,基因表达分析显示和mRNA水平下调以及miR-34a上调。此外,转染与处理后的HEC-1A细胞凋亡增加和迁移减少相关。
该研究强调了Snail-1对EC细胞中EMT的显著影响,从而影响凋亡和转移。通过特异性siRNA对Snail-1进行靶向沉默成为治疗EC的一种有前景的治疗方法。