Yu Hong, Sun Jianying, Jiang Shaochang, Xu Ying
Department of Gastroenterology, Yantai Yuhuangding Hospital, Yantai, Shandong 264000, P.R. China.
Department of Internal Medicine, Yantai Yuhuangding Hospital, Yantai, Shandong 264000, P.R. China.
Exp Ther Med. 2019 Feb;17(2):1330-1336. doi: 10.3892/etm.2018.7042. Epub 2018 Dec 3.
microRNA (miRNA) expression profiles of gastric cancer (GC) and adjacent healthy gastric mucosa tissue were used to search for differentially expressed miRNAs, identifying downregulated miRNA-490-3p in GC. The present study aimed to investigate the cellular function of miRNA-490-3p and its underlying mechanism in the occurrence and progression of GC. Reverse transcription-quantitative polymerase chain reaction was used to measure miRNA-490-3p expression levels in GC tissue and adjacent healthy tissue samples. The regulatory effect of miRNA-490-3p on the proliferation and apoptosis of GC cells was detected by cell counting kit-8, colony formation assay and flow cytometry. Bioinformatic methods were used to predict AKT1 as the target of miRNA-490-3p and this was verified by a dual-luciferase reporter assay. Furthermore, western blot analysis was used to measure protein expression of AKT1 in GC cells following overexpression or knockdown of miRNA-490-3p. The present study demonstrated that miRNA-490-3p expression was downregulated in GC tissue, compared with adjacent healthy tissue. In particular, miRNA-490-3p expression levels were significantly decreased in GC tissue samples from patients with advanced cancer (stage III+IV) compared with samples from patients with early-stage (stage I+II) cancer. Additionally, miRNA-490-3p expression levels were significantly decreased in GC tissue samples from patients whose tumor size was >3 cm, compared with those <3 cm. , downregulation of miRNA-490-3p promoted cell proliferation and suppressed apoptosis. In addition, rescue experiments demonstrated that overexpression of AKT1 partially reversed the effect of miRNA-490-3p on cell proliferation and apoptosis. The present study demonstrated that miRNA-490-3p regulated proliferation and apoptosis in gastric cancer cells via direct targeting of AKT1.
利用胃癌（GC）及相邻健康胃黏膜组织的微小RNA（miRNA）表达谱来寻找差异表达的miRNA，确定GC中miRNA-490-3p表达下调。本研究旨在探讨miRNA-490-3p的细胞功能及其在GC发生发展中的潜在机制。采用逆转录-定量聚合酶链反应检测GC组织和相邻健康组织样本中miRNA-490-3p的表达水平。通过细胞计数试剂盒-8、集落形成试验和流式细胞术检测miRNA-490-3p对GC细胞增殖和凋亡的调节作用。利用生物信息学方法预测AKT1为miRNA-490-3p的靶标，并通过双荧光素酶报告基因试验进行验证。此外，采用蛋白质印迹分析检测miRNA-490-3p过表达或敲低后GC细胞中AKT1的蛋白表达。本研究表明，与相邻健康组织相比，GC组织中miRNA-490-3p表达下调。特别是，晚期癌症（III+IV期）患者的GC组织样本中miRNA-490-3p表达水平显著低于早期癌症（I+II期）患者的样本。此外，肿瘤大小>3 cm的患者的GC组织样本中miRNA-490-3p表达水平显著低于肿瘤大小<3 cm的患者。miRNA-490-3p下调促进细胞增殖并抑制凋亡。此外，挽救实验表明，AKT1过表达部分逆转了miRNA-490-3p对细胞增殖和凋亡的影响。本研究表明，miRNA-490-3p通过直接靶向AKT1调节胃癌细胞的增殖和凋亡。
