Su Xiaojuan, Wang Binbin, Zhang Bo, Pan Shiwen
Department of Radiology, The Second Affiliated Hospital of Soochow University, No. 1055, Sanxiang Road, Gusu District, Suzhou, Jiangsu 215004, China.
Department of Radiotherapy & Oncology, The Second Affiliated Hospital of Soochow University, No. 1055, Sanxiang Road, Gusu District, Suzhou, Jiangsu 215004, China.
J Radiat Res. 2023 May 6;64(3):558-68. doi: 10.1093/jrr/rrad033.
The therapeutic outcomes of ovarian cancer (OVCA) patients are majorly limited by the development of acquired chemo/radioresistance and the lack of targeted therapies. Accumulating studies demonstrate that microRNAs are involved in tumorigenesis and radioresistance. This study aims to illustrate the role of miR-588 in the radioresistance of OVCA cells. The levels of miR-588 and mRNAs were detected by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). OVCA cell viability, proliferative, migratory and invasive capacities were evaluated by the cell counting kit-8 (CCK-8) assay, colony formation assay, wound healing assay and transwell assay, respectively. The luciferase activities of plasmids containing wild -type and mutant serine/arginine-rich splicing factor 6 (SRSF6) 3'-untranslated region in miR-588 silenced OVCA cells were detected by a luciferase reporter assay. We found that miR-588 was overexpressed in OVCA tissues and cells. Knockdown of miR-588 exerted an inhibitory effect on the proliferation, migration and invasion and strengthened the radiosensitivity of OVCA cells, whereas overexpression of miR-588 increased the radioresistance of OVCA cells. SRSF6 was verified to be targeted by miR-588 in OVCA cells. In addition, the expression level of miR-588 was negatively correlated with that of SRSF6 in OVCA clinical samples. Rescue assays indicated that SRSF6 knockdown reversed the effect of miR-588 inhibition of OVCA cells under radiation. Overall, miR-588 acts as an oncogene in OVCA and increases the radioresistance of OVCA cells by targeting SRSF6.
卵巢癌(OVCA)患者的治疗效果主要受到获得性化疗/放疗抗性的发展以及缺乏靶向治疗的限制。越来越多的研究表明,微小RNA参与肿瘤发生和放疗抗性。本研究旨在阐明miR-588在OVCA细胞放疗抗性中的作用。通过逆转录定量聚合酶链反应(RT-qPCR)检测miR-588和mRNA的水平。分别通过细胞计数试剂盒-8(CCK-8)测定、集落形成测定、伤口愈合测定和Transwell测定评估OVCA细胞的活力、增殖、迁移和侵袭能力。通过荧光素酶报告基因测定检测在miR-588沉默的OVCA细胞中含有野生型和突变型富含丝氨酸/精氨酸剪接因子6(SRSF6)3'-非翻译区的质粒的荧光素酶活性。我们发现miR-588在OVCA组织和细胞中过表达。敲低miR-588对OVCA细胞的增殖、迁移和侵袭具有抑制作用,并增强了其放射敏感性,而miR-588的过表达增加了OVCA细胞的放疗抗性。在OVCA细胞中验证SRSF6是miR-588的靶标。此外,在OVCA临床样本中,miR-588的表达水平与SRSF6的表达水平呈负相关。挽救实验表明,敲低SRSF6可逆转miR-588抑制对辐射下OVCA细胞的影响。总体而言,miR-588在OVCA中作为癌基因发挥作用,并通过靶向SRSF6增加OVCA细胞的放疗抗性。