Huang Lina, Shen Qinqin, Yu Kun, Yang Jie, Li Xiuxiu
Department of Gynecology, The Affiliated Women and Children's Hospital of Ningbo University, Ningbo, Zhejiang, China.
Department of Science and Education, The Affiliated Women and Children's Hospital of Ningbo University, Ningbo, Zhejiang, China.
Biotechnol Appl Biochem. 2025 Apr;72(2):340-354. doi: 10.1002/bab.2667. Epub 2024 Sep 19.
Cervical cancer (CC) poses a threat to human health. Enhancing pyroptosis can prevent the proliferation and epithelial-mesenchymal transition (EMT) of tumor cells. This study aims to reveal the candidates that modulate pyroptosis in CC. Accordingly, the common microRNAs (miRNAs/miRs) that were sponged by RBPMS antisense RNA 1 (RBPMS-AS1) and could target Phospholipase C-Like 1 (PLCL1) were intersected. The expression of PBPMS-AS1/miR-19a-3p (candidate miRNA)/PLCL1 was predicted in cervical squamous cell carcinoma (CESC), by which the expression location of RBPMS-AS1 and the binding between RBPMS-AS1/PLCL1 and miR-19a-3p were analyzed. The targeting relationship between RBPMS-AS1/PLCL1 and miR-19a-3p was confirmed by dual-luciferase reporter assay. After the transfection, cell counting kit-8 assay, colony formation assay, quantitative reverse transcription PCR, and Western blot were implemented for cell viability and proliferation analysis as well as gene and protein expression quantification analysis. Based on the results, RBPMS-AS1 and PLCL1 were lowly expressed, yet miR-19a-3p was highly expressed in CESC. RBPMS-AS1 overexpression diminished the proliferation and expressions of N-cadherin, vimentin, and miR-19a-3p, yet enhanced those of E-cadherin, PLCL1, and pyroptosis-relevant proteins (inteleukin-1β, caspase-1, and gasdermin D N-terminal). However, the above RBPMS-AS1 overexpression-induced effects were counteracted in the presence of miR-19a-3p. There also existed a targeting relationship and negative interplay between PLCL1 and miR-19a-3p. In short, RBPMS-AS1 sponges miR-19a-3p and represses the growth and EMT of CC cells via enhancing PLCL1-mediated pyroptosis.
宫颈癌（CC）对人类健康构成威胁。增强细胞焦亡可预防肿瘤细胞的增殖和上皮-间质转化（EMT）。本研究旨在揭示调节CC中细胞焦亡的候选分子。因此，对被RBPMS反义RNA 1（RBPMS-AS1）吸附且可靶向磷脂酶C样1（PLCL1）的常见微小RNA（miRNA/miR）进行了交集分析。预测了PBPMS-AS1/miR-19a-3p（候选miRNA）/PLCL1在宫颈鳞状细胞癌（CESC）中的表达，据此分析了RBPMS-AS1的表达位置以及RBPMS-AS1/PLCL1与miR-19a-3p之间的结合情况。通过双荧光素酶报告基因实验证实了RBPMS-AS1/PLCL1与miR-19a-3p之间的靶向关系。转染后，采用细胞计数试剂盒-8实验、集落形成实验、定量逆转录PCR和蛋白质免疫印迹法进行细胞活力和增殖分析以及基因和蛋白质表达定量分析。结果显示，RBPMS-AS1和PLCL1在CESC中低表达，而miR-19a-3p高表达。RBPMS-AS1过表达可降低N-钙黏蛋白、波形蛋白和miR-19a-3p的增殖及表达，但可增强E-钙黏蛋白、PLCL1和细胞焦亡相关蛋白（白细胞介素-1β、半胱天冬酶-1和gasdermin D N端）的表达。然而，在miR-19a-3p存在的情况下，上述RBPMS-AS1过表达诱导的效应被抵消。PLCL1与miR-19a-3p之间也存在靶向关系和负向相互作用。简而言之，RBPMS-AS1吸附miR-19a-3p，并通过增强PLCL1介导的细胞焦亡来抑制CC细胞的生长和EMT。
