Wang Gang, Lu Yu, Di Shi, Xie Maohua, Jing Fang, Dai Xiaoyan
Department of Gynecology, Wuhan Third Hospital (Tongren Hospital of Wuhan University), Wuhan, Hubei 430061, P.R. China.
Department of Vasculocardiology, Wuhan Wuchang Hospital, Wuhan, Hubei 430061, P.R. China.
Oncol Lett. 2022 May 27;24(1):228. doi: 10.3892/ol.2022.13349. eCollection 2022 Jul.
Cervical cancer (CC) is one of the most common gynecological malignancies that endangers women's health. A negative effect of glycolysis is that it contributes to abnormal tumor growth. MicroRNA (miR)-99a expression has been found to be decreased in CC. The present study aimed to investigate the role of miR-99a-5p in glycolysis in CC. For this purpose, the association between miR-99a and the prognosis of patients with CC from The Cancer Genome Atlas database was analyzed using Kaplan-Meier analysis. miR-99a-5p expression and Ras-related GTP binding D (RRAGD) expression were assessed using reverse transcription-quantitative PCR and western blot analysis. Cell proliferation and apoptosis were examined using an MTT assay and flow cytometry, respectively. Glucose uptake, lactate concentration and extracellular acidification rate were measured using a glucose uptake colorimetric assay, a lactate colorimetric assay and a Seahorse XFe96 extracellular flux analyzer, respectively. The association between miR-99a-5p and RRAGD was predicted using TargetScan 7.1, and was confirmed using dual luciferase reporter assay. The results revealed that miR-99a-5p expression was decreased and that of RRAGD was increased in CC tissues and cell lines. RRAGD was negatively regulated by miR-99a-5p. The overexpression of miR-99a-5p induced apoptosis and inhibited glycolysis in CC cells by targeting RRAGD. On the whole, the present study revealed a novel mechanism through which miR-99a-5p regulates cell apoptosis and glycolysis in CC, thus providing a potential therapeutic target for CC.
宫颈癌(CC)是危害女性健康的最常见妇科恶性肿瘤之一。糖酵解的一个负面影响是它会导致肿瘤异常生长。已发现微小RNA(miR)-99a在宫颈癌中表达降低。本研究旨在探讨miR-99a-5p在宫颈癌糖酵解中的作用。为此,使用Kaplan-Meier分析方法分析了来自癌症基因组图谱数据库的miR-99a与宫颈癌患者预后之间的关联。采用逆转录定量PCR和蛋白质免疫印迹分析评估miR-99a-5p表达和Ras相关GTP结合蛋白D(RRAGD)表达。分别使用MTT法和流式细胞术检测细胞增殖和凋亡情况。分别使用葡萄糖摄取比色法、乳酸比色法和海马XFe96细胞外通量分析仪测量葡萄糖摄取、乳酸浓度和细胞外酸化率。使用TargetScan 7.1预测miR-99a-5p与RRAGD之间的关联,并通过双荧光素酶报告基因检测进行验证。结果显示,在宫颈癌组织和细胞系中,miR-99a-5p表达降低,RRAGD表达升高。RRAGD受miR-99a-5p负调控。miR-99a-5p的过表达通过靶向RRAGD诱导宫颈癌细胞凋亡并抑制糖酵解。总体而言,本研究揭示了miR-99a-5p调控宫颈癌细胞凋亡和糖酵解的新机制,从而为宫颈癌提供了一个潜在的治疗靶点。