Rasheed Zafar
Department of Pathology, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
Int J Health Sci (Qassim). 2024 Nov-Dec;18(6):25-30.
Interleukin-8 (IL-8) and microRNA-183-5p (hsa-miR-183-5p) have been implicated in the development of cervical cancer, yet their relationship has not been explored. This study aims to determine whether phorbol 12-myristate 13-acetate (PMA)-induced IL-8 expression is regulated by hsa-miR-183-5p in cervical cancer cells.
Bioinformatics algorithms were employed to predict the potential binding of hsa-miR-183-5p to the 3'UTR of IL-8 mRNA. CaSKi cervical cancer cells were used as a model to investigate this regulation. The expression levels of hsa-miR-183-5p and IL-8 were measured using Taqman assays through real-time polymerase chain reaction, while IL-8 protein levels were quantified in culture media through IL-8 specific Sandwich enzyme-linked immunosorbent assays. Luciferase reporter assays and transfections with pre- or anti-miR-183-5p were conducted to validate the binding of hsa-miR-183-5p to IL-8 mRNA's 3'UTR.
The bioinformatics tool TargetScan identified a seed-matched sequence for hsa-miR-183-5p in the 3'UTR of IL-8 mRNA. PMA-induced IL-8 expression was inversely correlated with hsa-miR-183-5p down regulation in cervical cancer cells. hsa-miR-183-5p significantly reduced luciferase activity in the 3'UTR-IL-8 reporter assay. Transfection with pre-miR-183-5p led to a notable decrease in IL-8 mRNA and protein secretion, while anti-miR-183-5p transfection caused a significant increase in IL-8 mRNA and protein levels in PMA-treated cells.
This study is the first to demonstrate that hsa-miR-183-5p directly regulates IL-8 expression in cervical cancer cells. Both IL-8 and hsa-miR-183-5p could serve as potential therapeutic targets in the treatment of cervical cancer.
白细胞介素-8(IL-8)和微小RNA-183-5p(hsa-miR-183-5p)与宫颈癌的发生发展有关,但其关系尚未得到探究。本研究旨在确定佛波酯12-肉豆蔻酸酯13-乙酸酯(PMA)诱导的IL-8表达是否受hsa-miR-183-5p在宫颈癌细胞中的调控。
采用生物信息学算法预测hsa-miR-183-5p与IL-8 mRNA 3'非翻译区(3'UTR)的潜在结合。以CaSKi宫颈癌细胞为模型研究这种调控。通过实时聚合酶链反应使用Taqman分析测定hsa-miR-183-5p和IL-8的表达水平,同时通过IL-8特异性夹心酶联免疫吸附测定法定量培养基中的IL-8蛋白水平。进行荧光素酶报告基因测定以及用pre-或anti-miR-183-5p转染以验证hsa-miR-183-5p与IL-8 mRNA 3'UTR的结合。
生物信息学工具TargetScan在IL-8 mRNA的3'UTR中鉴定出hsa-miR-183-5p的种子匹配序列。在宫颈癌细胞中,PMA诱导的IL-8表达与hsa-miR-183-5p下调呈负相关。在3'UTR-IL-8报告基因测定中,hsa-miR-183-5p显著降低荧光素酶活性。用pre-miR-183-5p转染导致IL-8 mRNA和蛋白分泌显著减少,而anti-miR-183-5p转染导致PMA处理细胞中IL-8 mRNA和蛋白水平显著增加。
本研究首次证明hsa-miR-183-5p直接调控宫颈癌细胞中IL-8的表达。IL-8和hsa-miR-183-5p均可作为宫颈癌治疗的潜在靶点。