Lim Hyoung-Sup, Kim Chun Sung, Kim Jae-Sung, Yu Sun-Kyoung, Go Dae-San, Lee Seul Ah, Moon Sung Min, Chun Hong Sung, Kim Su Gwan, Kim DO Kyung
Oral Biology Research Institute, School of Dentistry, Chosun University, Gwangju, Republic of Korea.
Department of Biomedical Science, Chosun University, Gwangju, Republic of Korea.
Anticancer Res. 2017 Oct;37(10):5425-5433. doi: 10.21873/anticanres.11970.
BACKGROUND/AIM: The purpose of this study was to elucidate the molecular mechanism underlying regulation of semaphorin-6A (SEMA6A) involving microRNA-203 (miR-203) as a tumor suppressor in YD-38 human oral cancer cells.
miRNA arrays, polymerase chain reaction analyses, MTT assays, immunoblotting, and luciferase assays were carried out in YD-38 cells.
MiRNA microarray results showed that expression of miR-203 was significantly down-regulated in YD-38 cells compared to normal human oral keratinocytes. The viability of YD-38 cells was reduced by miR-203 in time- and dose-dependent manners. Overexpression of miR-203 increased the nuclear condensation of YD-38 cells and activated the apoptotic signaling pathway by up-regulating pro-apoptotic factors, such as BCL-2-associated X protein (BAX) and BCL-2 homologous antagonist killer (BAK), and the active forms of caspase-9, caspase-3, and poly-(ADP-ribose)-polymerase (PARP). Furthermore, target gene array analyses revealed that the expression of class 6 semaphorin A (SEMA6A) was down-regulated by miR-203 in YD-38 cells. Both the mRNA and protein levels of SEMA6A were reduced in YD-38 cells transfected with miR-203. Luciferase activity assay confirmed that miR-203 directly targets the SEMA6A 3'-untranslated region to suppress gene expression.
Our results indicate that miR-203 induces the apoptosis of YD-38 human oral cancer cells by directly targeting SEMA6A, suggesting its potential application in anticancer therapeutics.
背景/目的：本研究旨在阐明在YD-38人舌癌细胞中，作为肿瘤抑制因子的微小RNA-203（miR-203）参与调控信号素-6A（SEMA6A）的分子机制。
对YD-38细胞进行了miRNA芯片、聚合酶链反应分析、MTT检测、免疫印迹和荧光素酶检测。
miRNA芯片结果显示，与正常人口腔角质形成细胞相比，YD-38细胞中miR-203的表达显著下调。miR-203以时间和剂量依赖的方式降低了YD-38细胞的活力。miR-203的过表达增加了YD-38细胞的核浓缩，并通过上调促凋亡信号通路相关蛋白，如BCL-2相关X蛋白（BAX）、BCL-2同源拮抗剂杀手蛋白（BAK）以及半胱天冬酶-9、半胱天冬酶-3和聚（ADP-核糖）聚合酶（PARP）的活性形式，激活了凋亡信号通路。此外，靶基因芯片分析显示，在YD-38细胞中，miR-203下调了6类信号素A（SEMA6A）的表达。在转染了miR-203的YD-38细胞中，SEMA6A的mRNA和蛋白水平均降低。荧光素酶活性检测证实，miR-203直接靶向SEMA6A的3'-非翻译区以抑制基因表达。
我们的结果表明，miR-203通过直接靶向SEMA6A诱导YD-38人舌癌细胞凋亡，提示其在抗癌治疗中的潜在应用价值。
