Gastric cancer(GC)was referred to a malignant tumor of the digestive tract originating from the epithelium of gastric mucosa.Transcription factor DLX5 was verified as an oncogene in various types of tumors,while miR-3...Gastric cancer(GC)was referred to a malignant tumor of the digestive tract originating from the epithelium of gastric mucosa.Transcription factor DLX5 was verified as an oncogene in various types of tumors,while miR-376a-3p was speculated as a tumor suppressor.Based on the bioinformatics database,we hypothesized that miR-376a participated in the regulation of GC development by targeting DLX5.Compared with adjacent tissue,a significant increase of DLX5 expression was determined in GC tissues,but the expression level is significantly reduced in miR-376a.Similar expression signature of DLX5 and miR-376a was also determined between 4 GC cells(HGC,SGC,MGC,and AGS cell lines)and GES cell line.The level of DLX5 was notably reduced in HGC and MGC cell lines after miR-376a-3p overexpression,and increased after miR-376a-3p inhibition.Then,the inhibition role of miR-376a-3p on DLX5 was further proved by dualluciferase reporter assay.Gain-of-function experiments showed that upregulation of miR-376a-3p in GC cells could inhibit the ability of epithelial-mesenchymal transition,proliferation,and invasion,and enhance the GC cell apoptosis level.However,these roles of miR-376a-3p could be abolished by DLX5 overexpression.This study confirmed that reduction of miR-376a-3p expression level in GC cells would lead to the increase in cell growth and invasion,indicating that upregulation of miR-376a-3p might have a potential therapeutic role on GC.展开更多
背景:目前已有针对miRNA/mRNA轴调节骨关节炎疾病进程的分子机制研究。先前生物信息学研究发现具有临床预测价值的mRNA(磷脂酶Cδ3:phospholipase C delta 3,PLCD3)及其靶向miRNA(miR-34a-5p),尚缺实验验证其调控骨关节炎的具体作用及...背景:目前已有针对miRNA/mRNA轴调节骨关节炎疾病进程的分子机制研究。先前生物信息学研究发现具有临床预测价值的mRNA(磷脂酶Cδ3:phospholipase C delta 3,PLCD3)及其靶向miRNA(miR-34a-5p),尚缺实验验证其调控骨关节炎的具体作用及机制。目的:探讨miR-34a-5p/PLCD3轴对骨关节炎进展的调控作用及机制。方法:选择15例膝骨关节炎患者的滑膜为骨关节炎组,同时选择同期因创伤致髌骨骨折行内固定术的15例年轻患者的健康滑膜为对照组,Real-time PCR法检测滑膜中PLCD3及miR-34a-5p的表达。通过细胞转染的方法,将人滑膜关节炎成纤维细胞(human fibroblast like synovial cells-osteoarthritis,HFLS-OA)进行处理,并分为miR-34a-5p模拟物组、pCDH-PLCD3组、miR-34a-5p模拟物+pCDH-PLCD3组、miR-34a-5p抑制剂组、si-PLCD3组、miR-34a-5p抑制剂+si-PLCD3组,通过Real-time PCR法检测PLCD3和miR-34a-5p表达的关系;通过CCK-8法、细胞划痕实验检测各组HFLS-OA细胞活力及细胞迁移的影响;使用Western Blot法检测凋亡标记蛋白表达水平;使用ELISA法检测炎症因子的表达。结果与结论:①PLCD3是miR-34a-5p的直接靶标,同时PLCD3和miR-34a-5p表达水平呈负相关。②PLCD3上调会促进HFLS-OA细胞的增殖并抑制细胞迁移,而miR-34a-5p上调会显著抑制HFLS-OA细胞的活性并增强细胞迁移;miR-34a-5p过表达使HFLS-OA细胞Casp3和Casp9蛋白水平显著升高,而PLCD3过表达则表现出相反趋势。③PLCD3过表达显著增加了HFLS-OA细胞白细胞介素6和肿瘤坏死因子α的表达,而miR-34a-5p模拟物则表现出保护活性。④结果说明,miR-34a-5p/PLCD3轴可能通过调节滑膜细胞的炎症过程或凋亡来影响骨关节炎的进展。展开更多
基金This work was supported by the Nature Science Foundation of China(81972320).
文摘Gastric cancer(GC)was referred to a malignant tumor of the digestive tract originating from the epithelium of gastric mucosa.Transcription factor DLX5 was verified as an oncogene in various types of tumors,while miR-376a-3p was speculated as a tumor suppressor.Based on the bioinformatics database,we hypothesized that miR-376a participated in the regulation of GC development by targeting DLX5.Compared with adjacent tissue,a significant increase of DLX5 expression was determined in GC tissues,but the expression level is significantly reduced in miR-376a.Similar expression signature of DLX5 and miR-376a was also determined between 4 GC cells(HGC,SGC,MGC,and AGS cell lines)and GES cell line.The level of DLX5 was notably reduced in HGC and MGC cell lines after miR-376a-3p overexpression,and increased after miR-376a-3p inhibition.Then,the inhibition role of miR-376a-3p on DLX5 was further proved by dualluciferase reporter assay.Gain-of-function experiments showed that upregulation of miR-376a-3p in GC cells could inhibit the ability of epithelial-mesenchymal transition,proliferation,and invasion,and enhance the GC cell apoptosis level.However,these roles of miR-376a-3p could be abolished by DLX5 overexpression.This study confirmed that reduction of miR-376a-3p expression level in GC cells would lead to the increase in cell growth and invasion,indicating that upregulation of miR-376a-3p might have a potential therapeutic role on GC.
