Objective MiRNAs are closely related to tumors,and we hypothesized there is specific miR expression in nasopharyngeal carcinoma(NPC).We intended to investigate the expression of mir-34c-5p and mir-150-5p in NPC and to...Objective MiRNAs are closely related to tumors,and we hypothesized there is specific miR expression in nasopharyngeal carcinoma(NPC).We intended to investigate the expression of mir-34c-5p and mir-150-5p in NPC and to investigate the effects of mir-34c-5p and mir-150-5p on apoptosis and invasion following up-regulated expression in HNE1 NPC cells.Methods MiR-34c-5p and miR-150-5p expression levels in 30 individual cases of NPC and nasopharyngitis were detected with gene chip and qRT-PCR techniques.miR-34c-5p and miR-150-5p were transfected into the NPC cell line HNE1 via liposomes.Their expression levels were detected with qRT-PCR,apoptosis was evaluated by flow cytometry,and invasion ability was assessed via Transwell migration assay.Results MiR-150-5p expression levels in NPC and nasopharyngitis were 0.165±0.092 and 1.062±0.280 respectively,and miR-34c-5p expression levels in NPC and nasopharyngitis were 0.417±0.220 and 1.385±0.739,respectively,which indicated miR-34c-5p and miR-150-5p were weakly expressed in NPC.Apoptosis rates in HNE1 cells transfected by miR-34c-5p and miR-150-5p were increased,by 12.7%and 7.6%,respectively,which were significantly higher compared to blank control(3.9%).The Transwell assay demonstrated that invasive HNE1 cell counts were 32.00±2.00 and 28.33±2.08,respectively,compared to 60.66±8.50 in the blank control(P<0.001).Conclusion MiR-34c-5p and miR-150-5p are lowly expressed in NPC,and their down-regulation may be associated with NPC.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Supported by grants from the National Natural Science foundation of China(No.81373698)Joint Special Scientific Research Project of the Department of Science and Technology of Guangdong Province-Guangdong Provincial Academy of Chinese Medical Sciences(No.2017A020213020)Zhongshan Key Scientific and Technological Project(No.2015B1003,2019B1096)。
文摘Objective MiRNAs are closely related to tumors,and we hypothesized there is specific miR expression in nasopharyngeal carcinoma(NPC).We intended to investigate the expression of mir-34c-5p and mir-150-5p in NPC and to investigate the effects of mir-34c-5p and mir-150-5p on apoptosis and invasion following up-regulated expression in HNE1 NPC cells.Methods MiR-34c-5p and miR-150-5p expression levels in 30 individual cases of NPC and nasopharyngitis were detected with gene chip and qRT-PCR techniques.miR-34c-5p and miR-150-5p were transfected into the NPC cell line HNE1 via liposomes.Their expression levels were detected with qRT-PCR,apoptosis was evaluated by flow cytometry,and invasion ability was assessed via Transwell migration assay.Results MiR-150-5p expression levels in NPC and nasopharyngitis were 0.165±0.092 and 1.062±0.280 respectively,and miR-34c-5p expression levels in NPC and nasopharyngitis were 0.417±0.220 and 1.385±0.739,respectively,which indicated miR-34c-5p and miR-150-5p were weakly expressed in NPC.Apoptosis rates in HNE1 cells transfected by miR-34c-5p and miR-150-5p were increased,by 12.7%and 7.6%,respectively,which were significantly higher compared to blank control(3.9%).The Transwell assay demonstrated that invasive HNE1 cell counts were 32.00±2.00 and 28.33±2.08,respectively,compared to 60.66±8.50 in the blank control(P<0.001).Conclusion MiR-34c-5p and miR-150-5p are lowly expressed in NPC,and their down-regulation may be associated with NPC.
