At present,the role of many long non-coding RNAs(lncRNAs)as tumor suppressors in the formation and development of cervical cancer(CC)has been studied.However,lncRNA prostate cancer gene expression marker 1(PCGEM1),who...At present,the role of many long non-coding RNAs(lncRNAs)as tumor suppressors in the formation and development of cervical cancer(CC)has been studied.However,lncRNA prostate cancer gene expression marker 1(PCGEM1),whose high expression not only aggravates ovarian cancer but also can induce tumorigenesis and endometrial cancer progression,has not been studied in CC.The objective of this study was to investigate the expression and the underlying role of PCGEM1 in CC.The relative expression of PCGEM1 in CC cells was detected by real-time PCR.After the suppression of PCGEM1 expression by shRNA,the changes in the proliferation,migration,and invasion capacities were detected via CCK-8 assay,EdU assay,and colony formation assay wound healing assay.Transwell assay and the changes in expressions of epithelial-to-mesenchymal transition(EMT)markers were determined by western blot and immunofluorescence.The interplay among PCGEM1,miR-642a-5p,and kinesin family member 5B(KIF5B)was confirmed by bioinformatics analyses and luciferase reporter assay.Results showed that PCGEM1 expressions were up-regulated within CC cells.Cell viabilities,migration,and invasion were remarkably reduced after the suppression of PCGEM1 expression by shRNA in Hela and SiHa cells.N-cadherin was silenced,but E-cadherin expression was elevated by sh-PCGEM1.Moreover,by sponging miR-642a-5p in CC,PCGEM1 was verified as a competitive endogenous RNA(ceRNA)that modulates KIF5B levels.MiR-642a-5p down-regulation partially rescued sh-PCGEM1’s inhibitory effects on cell proliferation,migration,invasion,and EMT process.In conclusion,the PCGEM1/miR-642a-5p/KIF5B signaling axis might be a novel therapeutic target in CC.This study provides a research basis and new direction for targeted therapy of CC.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the Nantong Municipal Health Commission Research Project(MB2021054)for this study.
文摘At present,the role of many long non-coding RNAs(lncRNAs)as tumor suppressors in the formation and development of cervical cancer(CC)has been studied.However,lncRNA prostate cancer gene expression marker 1(PCGEM1),whose high expression not only aggravates ovarian cancer but also can induce tumorigenesis and endometrial cancer progression,has not been studied in CC.The objective of this study was to investigate the expression and the underlying role of PCGEM1 in CC.The relative expression of PCGEM1 in CC cells was detected by real-time PCR.After the suppression of PCGEM1 expression by shRNA,the changes in the proliferation,migration,and invasion capacities were detected via CCK-8 assay,EdU assay,and colony formation assay wound healing assay.Transwell assay and the changes in expressions of epithelial-to-mesenchymal transition(EMT)markers were determined by western blot and immunofluorescence.The interplay among PCGEM1,miR-642a-5p,and kinesin family member 5B(KIF5B)was confirmed by bioinformatics analyses and luciferase reporter assay.Results showed that PCGEM1 expressions were up-regulated within CC cells.Cell viabilities,migration,and invasion were remarkably reduced after the suppression of PCGEM1 expression by shRNA in Hela and SiHa cells.N-cadherin was silenced,but E-cadherin expression was elevated by sh-PCGEM1.Moreover,by sponging miR-642a-5p in CC,PCGEM1 was verified as a competitive endogenous RNA(ceRNA)that modulates KIF5B levels.MiR-642a-5p down-regulation partially rescued sh-PCGEM1’s inhibitory effects on cell proliferation,migration,invasion,and EMT process.In conclusion,the PCGEM1/miR-642a-5p/KIF5B signaling axis might be a novel therapeutic target in CC.This study provides a research basis and new direction for targeted therapy of CC.
