MiR-542-5p is a negative prognostic factor and promotes osteosarcoma tumorigenesis by targeting HUWE1

Recent evidence has demonstrated that microRNAs (miRNAs) are involved in the proliferation and metastasis of osteosarcoma. Using miRNA microarray and functional screening methods to compare miRNA expression profiles in osteosarcoma cell lines treated with Trichostatin A (TSA), overexpression of miR-542-5p was determined to be involved in the proliferation of osteosarcoma. We used isobaric tags for relative and absolute quantitation (iTRAQ) and nanoscale liquid chromatography-mass spectrometry (NanoLC−MS/MS) to identify differentially expressed proteins in MNNG/HOS and U2OS osteosarcoma cell lines transfected with miR-542-5p; in both cell lines, seven proteins were downregulated, and nine were upregulated. HUWE1 was found to be a direct target of miR-542-5p in both osteosarcoma cell lines, and was negatively correlated with miR-542-5p levels in human osteosarcoma tissues. Moreover, the expression of miR-542-5p was upregulated in human osteosarcoma tissue compared with non-tumor adjacent tissue. Kaplan-Meier analysis revealed that overexpression of miR-542-5p predicted poor prognosis for osteosarcoma patients. Taken together, our results indicated that miR-542-5p plays a critical role in the proliferation of osteosarcoma and targets HUWE1.     

LncRNA-HOTAIRM1 promotes aerobic glycolysis and proliferation in osteosarcoma via the miR-664b-3p/Rheb/mTOR pathway

Osteosarcoma (OS), which is a common and aggressive primary bone malignancy, occurs mainly in children and adolescent. Long noncoding RNAs (lncRNAs) are reported to play a pivotal role in various cancers. Here, we found that the lncRNA HOTAIRM1 is upregulated in OS cells and tissues. A set of functional experiments suggested that HOTAIRM1 knockdown attenuated the proliferation and stimulated the apoptosis of OS cells. A subsequent mechanistic study revealed that HOTAIRM1 functions as a competing endogenous RNA to elevate ras homologue enriched in brain (Rheb) expression by sponging miR-664b-3p. Immediately afterward, upregulated Rheb facilitates proliferation and suppresses apoptosis by promoting the mTOR pathway-mediated Warburg effect in OS. In summary, our findings demonstrated that HOTAIRM1 promotes the proliferation and suppresses the apoptosis of OS cells by enhancing the Warburg effect via the miR-664b-3p/Rheb/mTOR axis. Understanding the underlying mechanisms and targeting the HOTAIRM1/miR-664b-3p/Rheb/mTOR axis are essential for OS clinical treatment.     

LINC01123 enhances osteosarcoma cell growth by activating the Hedgehog pathway via the miR-516b-5p/Gli1 axis

The lncRNA LINC01123 has been reported to act as an oncogene in many human cancers. Nevertheless, the function and underlying mechanism of LINC01123 in osteosarcoma (OS) remain unclear. This study aimed to explore the roles and mechanisms of LINC01123 in OS progression. In this study, the expression of LINC01123 was significantly upregulated in OS cell lines than in a human osteoblast cell line. Furthermore, in vitro and in vivo experiments confirmed that knockdown of LINC01123 suppressed cell progression. Mechanistically, LINC01123 acted as a competing endogenous RNA by sponging miR-516b-5p, thus, increasing Gli1 expression by directly targeting its 3ʹUTR. Taken together, LINC01123 enhances OS proliferation and metastasis via the miR-516b-5p/Gli1 axis. Therefore, LINC01123 may be a potential therapeutic target for OS treatment.     

Linc00460通过海绵吸附影响miR-320a对乳腺癌细胞的有氧糖酵解作用

目的 探讨Linc00460通过海绵吸附miR-320a对乳腺癌（BC）细胞有氧糖酵解作用的影响。方法 qRT-PCR法检测正常乳腺上皮细胞系MCF-10A及5种BC细胞系中Linc00460和miR-320a表达水平。qRT-PCR检测干扰Linc00460对miR-320a表达的影响,双荧光素酶报告基因实验检测miR-320a和Linc00460的靶向关系。将si-Linc00460和miR-320a inhibitor共转染至MDA-MB-231细胞,qRT-PCR检测细胞中miR-320a表达水平;MTT法检测细胞增殖能力;2-NBDG法检测细胞葡萄糖摄取率;比色法检测细胞上清液中乳酸含量;酶活性试剂盒检测糖酵解关键酶的活性;Western blot检测酵解途径中关键蛋白表达水平。结果 与MCF-10A细胞比较,5种BC细胞系中Linc00460高表达,而miR-320a低表达。干扰Linc00460后,MDA-MB-231细胞中miR-320a表达显著升高。双荧光素酶报告基因实验证实,miR-320a和Linc00460可靶向结合。干扰Linc00460表达后MDA-MB-231细胞增殖能力受到明显抑制（P=0.000）,细胞葡萄糖摄取率和细胞上清液中乳酸含量降低（均P=0.000）,PFK、PK和LDH酶活性受到抑制（均P=0.000）,PFKM、GLUT1和LDHA蛋白表达水平下调（均P=0.000）。抑制miR-320a可明显逆转si-Linc00460对MDA-MB-231细胞增殖和糖酵解的抑制作用（均P=0.000或0.001）。结论 Linc00460可能通过海绵吸附miR-320a,上调PFKM表达,从而促进BC细胞有氧糖酵解作用。     

LPLUNC1 reduces glycolysis in nasopharyngeal carcinoma cells through the PHB1-p53/c-Myc axis

Cancer cells prefer glycolysis to support their proliferation. Our previous studies have shown that the long palate, lung, and nasal epithelial cell clone 1 (LPLUNC1) can upregulate prohibitin 1 (PHB1) expression to inhibit the proliferation of nasopharyngeal carcinoma (NPC) cells. Given that PHB1 is an important regulator of cell energy metabolism, we explored whether and how LPLUNC1 regulated glucose glycolysis in NPC cells. LPLUNC1 or PHB1 overexpression decreased glycolysis and increased oxidative phosphorylation (OXPHOS)-related protein expression in NPC cells, promoting phosphorylated PHB1 nuclear translocation through 14-3-3σ. LPLUNC1 overexpression also increased p53 but decreased c-Myc expression in NPC cells, which were crucial for the decrease in glycolysis and increase in OXPHOS-related protein expression induced by LPLUNC1 overexpression. Finally, we found that treatment with all-trans retinoic acid (ATRA) reduced the viability and clonogenicity of NPC cells, decreased glycolysis, and increased OXPHOS-related protein expression by enhancing LPLUNC1 expression in NPC cells. Therefore, the LPLUNC1-PHB1-p53/c-Myc axis decreased glycolysis in NPC cells, and ATRA upregulated LPLUNC1 expression, ATRA maybe a promising drug for the treatment of NPC.     

lncRNA MAFG-AS1通过调控miR-11181-3p/GLG1轴促进胃癌AGS细胞迁移、侵袭和有氧糖酵解

目的：探讨lncRNA MAFG-AS1/miR-11181-3p/GLG1分子轴对胃癌（gastric cancer,GC）细胞迁移、侵袭和有氧糖酵解的影响及其可能的机制。方法：选取 MAFG-AS1 相对高表达的 GC 细胞系 AGS 作为研究对象,采用 qPCR 法检测其 MAFGAS1、miR-11181-3p、GLG1的RNA表达水平,Transwell实验、糖酵解分析等检测细胞迁移、侵袭和有氧糖酵解的变化,利用生物信息学分析及双荧光素酶报告基因验证MAFG-AS1、miR-11181-3p、GLG1之间的相互作用关系。结果：敲减MAFG-AS1显著上调miR-11181-3p及下调GLG1的表达（均P<0.01）,并可显著抑制GC细胞迁移、侵袭和有氧糖酵解（均P<0.01）;荧光素酶报告基因证实MAFG-AS1竞争性吸附miR-11181-3p（P<0.01）;抑制miR-11181-3p或过表达GLG1可部分逆转敲减MAFG-AS1对GC细胞迁移、侵袭和有氧糖酵解的抑制作用（均P<0.05或P<0.01）。结论：MAFG-AS1通过miR-11181-3p/GLG1分子轴增强GC迁移、侵袭和有氧糖酵解,可能是GC诊疗的潜在分子靶点。     

lncRNAXIST/miR-34a-5p/SIRT6 分子轴调控口腔鳞癌细胞的增殖及转移

[摘要] 目的：探究lncRNA XIST/miR-34a-5p/SIRT6 分子轴调控口腔鳞癌细胞增殖和转移及其分子机制。方法：收集2013年3 月至2018 年3 月在青岛市口腔医院就诊的OSCC患者47 例癌组织和癌旁组织标本,采用qPCR检测OSCC患者组织及细胞系中lncRNA XIST、miR-34a-5p、SIRT6 mRNA 的表达,WB检测OSCC 患者组织及细胞系中SIRT6、Ki67、pcDNA、cleaved-caspase3、cleaved-caspase8、E-cadherin、Vimentin 蛋白的表达,采用CCK-8 实验检测敲降lncRNA XIST对Cal-27 及Tca-8113 细胞增殖的影响,Transwell 小室法检测Cal-27 及Tca-8113 细胞迁移及侵袭;流式细胞术检测Cal-27 及Tca-8113 细胞凋亡情况,双荧光素酶报告基因检测lncRNA XIST与miR-34a-5p、miR-34a-5p 与SIRT6 靶向结合关系。结果：lncRNA XIST和SIRT6 在OSCC患者癌组织及细胞系中高表达（均P<0.05）,miR-34a-5p 则呈低表达（P<0.01）;敲降lncRNA XIST抑制OSCC细胞的增殖、迁移及侵袭并促进细胞凋亡（均P<0.01）,同时转染miR-34a-5p 抑制剂或pcDNA-SIRT6 载体作用则相反;敲降lncRNA XIST 促进OSCC细胞中增殖及转移相关蛋白表达（均P<0.01）,同时转染miR-34a-5p 抑制剂或pcDNA-SIRT6 载体作用则相反;lncRNA XIST 与miR-34a 靶向结合,miR-34a 与SIRT6 靶向结合;lncRNA XIST 通过靶向miR-34a-5p 上调SIRT6 表达（P<0.01）。结论：lncRNA XIST/miR-34a-5p/SIRT6 分子轴能够调控OSCC细胞增殖及转移,为OSCC治疗提供潜在靶点。     

lncRNA TPTEP1通过抑制miR-129-5p影响膀胱癌T24细胞的增殖和侵袭

目的:分析长链非编码RNA(long-chain non-coding RNA,lncRNA) TPTEPl在膀胱癌组织和细胞的表达,观察其对膀胱癌细胞增殖和侵袭的影响及其作用机制.方法:收集2017年8月至2019年10月在武汉市东西湖人民医院泌尿外科接受手术治疗的43例膀胱癌患者的癌与癌旁组织标本,采用实时荧光定量...     

lncRNA XIST调控miR-486-5p影响食管癌细胞增殖的实验研究

目的:探讨lncRNA XIST在食管癌组织中的表达情况以及其对食管癌细胞增殖的影响。方法:实时定量PCR检测23对食管癌及癌旁组织中lncRNA XIST的表达水平。采用lncRNA XIST siRNA转染lncRNA XIST表达最高的两株食管癌细胞，实时定量PCR检测转染效率；CCK-8检测细胞增殖情况；双荧光素酶报告基因检测lncRNA XIST与miR-486-5p的结合情况；实时定量PCR进一步检测miR-486-5p的表达。结果:实时定量PCR结果显示与癌旁组织相比食管癌组织中lncRNA XIST的表达水平显著升高（P＜0.01）。此外，在三株食管癌细胞中lncRNA XIST在Eca-109和TE-1细胞中表达最高。在Eca-109和TE-1细胞中转染lncRNA XIST siRNA后lncRNA XIST的表达水平分别降低了（75.50±0.89）%和（64.74±13.42）%。下调lncRNA XIST显著抑制食管癌细胞的增殖。双荧光素酶报告基因检测结果显示wt-XIST和miR-486-5p mimic共转染的细胞荧光素酶活性显著降低（P＜0.01）。此外，下调lncRNA XIST后两株细胞中miR-486-5p的表达水平均显著升高（P＜0.01）。结论:lncRNA XIST与食管癌的恶性进展密切相关，lncRNA XIST有望成为食管癌治疗的新靶点。     

长链非编码RNA HOXD-AS1介导骨肉瘤细胞阿霉素耐药的机制研究

目的:观察长链非编码RNA HOXD-AS1对骨肉瘤(osteosarcoma,OS)细胞增殖、凋亡能力以及对阿霉素化疗耐药的影响，并探讨其机制。方法:首先采用实时荧光聚合酶链反应法(real-time polymerase chain reaction，RT-PCR)检测人正常成骨细胞和OS细胞系以及阿霉素耐药OS细胞系中lncRNA HOXD cluster antisense RNA 1 (lncRNA HOXD-AS1)和P-gp（P-glycoprotein）的表达水平；在骨肉瘤细胞中抑制lncRNA HOXD-AS1表达后，通过RT-PCR和Western-blot检测P-gp的表达水平；在骨肉瘤细胞中抑制lncRNA HOXD-AS1表达后，再过表达P-gp，应用CCK-8 法检测转染后各组吸光值以评价增殖率，应用Annexin V/PI染色流式细胞术检测各组细胞凋亡率，在培养基中加入阿霉素检测各组细胞存活率。结果:与正常成骨细胞相比，OS细胞中lncRNA HOXD-AS1过表达，P-gp过表达(P＜0.05)；抑制lncRNA HOXD-AS1表达后OS细胞中P-gp表达下调(P＜0.05)；OS细胞中抑制lncRNA HOXD-AS1表达后细胞增殖转移能力明显减弱（P＜0.05），凋亡增加（P＜0.05），化疗耐药减弱（P＜0.05）；而过表达P-gp后恶性表型明显恢复（P＜0.05）。结论:LncRNA HOXD-AS1可通过调控 P-gp的表达而调节OS细胞的增殖、凋亡和化疗耐药，参与OS的发生发展。     

LncRNA HOXC-AS3通过miR-15b-5p/E2F3生物学轴促进胃癌细胞的增殖和迁移

背景与目的：胃癌是一种发病率和死亡率均较高的恶性肿瘤。长链非编码RNA（long non-coding RNA,lncRNA）是一类具有调控功能的大分子非编码RNA,在胃癌的转移中发挥着重要的作用。探讨lncRNA HOXC-AS3（lnc-HOXC-AS3）在胃癌中的表达模式,以及通过miR-15b-5p/E2F3生物学轴促进胃癌细胞增殖和转移的分子机制。方法：根据生物信息学分析的方法寻找收集2017年4月—2018年12月安徽医科大学第一附属医院收治并经病理学检查诊断为胃癌的90例患者的胃癌组织中异常表达的lncRNA,并且利用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR）检测lnc-HOXC-AS3的表达水平;在对lnc-HOXC-AS3功能的研究中,采用细胞计数试剂盒（cell counting kit-8,CCK-8）和transwell等方法确定了lnc-HOXC-AS3对胃癌细胞（MGC803）增殖和迁移的影响;在机制上,采用双荧光素酶报告基因检测lnc-HOXC-AS3、miR-15b-5p和E2F3的靶向调控关系。结果：Lnc-HOXC-AS3在胃癌组织和胃癌细胞系中异常高表达。功能上,lnc-HOXC-AS3促进胃癌细胞增殖和迁移,相反,敲低lnc-HOXC-AS3则明显抑制胃癌细胞增殖和迁移。在机制的探讨中,通过双荧光素酶报告基因和一系列体外实验证实lnc-HOXC-AS3通过靶向下调miR-15b-5p的表达,进而解除miR-15b-5p对E2F3的抑制作用,促进了胃癌细胞增殖和迁移。结论：Lnc-HOXC-AS3在胃癌中异常高表达,并通过调控miR-15b-5p/E2F3生物学轴促进胃癌细胞增殖和迁移,渴望成为研究胃癌早期诊断和治疗的靶点。     

Elevated TRIM23 expression predicts cisplatin resistance in lung adenocarcinoma

The tripartite motif containing 23 (TRIM23) gene is a member of the tripartite motif (TRIM) family that participates in many pathophysiological processes. However, the role of TRIM23 in lung adenocarcinoma (LUAD) remains unclear. In the present study, TRIM23 was first screened by next‐generation sequencing between the cisplatin (DDP)‐resistant A549/DDP cell line and the parental A549 cell line, combined with integrated analysis of the Gene Expression Omnibus (GEO) data (E‐GEOD‐43493 and E‐GEOD‐43494). The expression of TRIM23 was then verified to be upregulated in the DDP‐resistant LUAD cells and tissues. The knockdown of TRIM23 expression in A549/DDP cells caused increased apoptosis, decreased IC₅₀ values of DDP, NF‐κB nuclear translocation, inhibition of cell proliferation in vitro and in vivo, inhibition of GLUT1/3 expression, glucose uptake, and lactate and ATP production. TRIM23 overexpression resulted in the opposite effects in A549 cells. In addition, the inhibition of proliferation in A549 cells caused by NF‐κB signaling inhibitor PTDC or glycolysis inhibitor 3‐BrPA could be weakened by TRIM23 overexpression. Furthermore, immunohistochemical analysis revealed that TRIM23 was upregulated in 46.1% (70/152) of LUAD cases, and elevated TRIM23 expression was correlated with high expression of NF‐κB, poor cellular differentiation, and adverse overall survival (OS) and disease‐free survival (DFS). In conclusion, our study demonstrates that TRIM23 acts as an oncogene in LUAD and promotes DDP resistance by regulating glucose metabolism via the TRIM23/NF‐κB/ GLUT1/3 axis.     

MicroRNA-mediated upregulation of integrin-linked kinase promotes Src-induced tumor progression

The tyrosine kinase c-Src is upregulated in various human cancers; however, the molecular mechanisms underlying c-Src-mediated tumor progression remain unclear. Here we show that downregulation of microRNA (miR)-542-3p is tightly associated with tumor progression via c-Src-related oncogenic pathways. In c-Src-transformed fibroblasts and human cancer cells that overexpress c-Src, miR-542-3p is substantially downregulated, and the ectopic expression of miR-542-3p suppresses tumor growth. We identified the integrin-linked kinase (ILK) as a conserved target of miR-542-3p. ILK upregulation promotes cell adhesion and invasion by activating the integrin-focal adhesion kinase (FAK)/c-Src pathway, and can also contribute to tumor growth via the AKT and glycogen synthase kinase 3β pathways. MiR-542-3p expression is downregulated by the activation of c-Src-related signaling molecules, including epidermal growth factor receptor, K-Ras and Ras/Raf/mitogen-activated protein kinase/extracellular signal-regulated kinase. In human colon cancer tissues, downregulation of miR-542-3p is significantly correlated with the upregulation of c-Src and ILK. Our results suggest that the novel c-Src-miR-542-3p-ILK-FAK circuit plays a crucial role in controlling tumor progression.     

lncRNA MEG3 通过miR-9-5p/SOCS5 轴对宫颈癌细胞恶性生物学行为的影响

目的：探讨lncRNA 母源性印记基因3（maternal imprinting gene 3, MEG3）通过miR-9-5p/SOCS5 轴对宫颈癌细胞增殖、迁移、侵袭和上皮间质转化（epithelial-mesenchymal transition, EMT）的调控作用。方法: 收集2017 年1 月至2019 年6 月重庆市中医院手术切除的20 例宫颈癌患者的癌及癌旁组织标本;利用脂质体转染技术分别将pcDNA3.1-MEG3、si-MEG3、miR-9-5p mimics、miR-9-5p inhibitor 及其对照质粒等转染进宫颈癌HeLa 和SiHa 细胞,构建过表达和沉默细胞模型。用qPCR检测宫颈癌组织及细胞模型中MEG3、miR-9-5p 和SOCS5 表达水平,用CCK-8 法、Transwell 小室法检测细胞的增殖、迁移和侵袭能力,用细胞免疫荧光实验检测细胞中E-cadherin 和vimentin 表达水平。通过在线生物信息学TargetScan 数据库预测靶基因,用双荧光素酶报告基因实验验证miR-9-5p 分别与MEG3 和SOCS5 的靶向关系。结果: 分别与癌旁组织和宫颈上皮HcerEpic 细胞比较,宫颈癌组织和细胞系中MEG3 和SOCS5 表达显著下调、miR-9-5p 表达显著上调（均P<0.01）。TargetScan 数据库分析和双荧光素酶报告基因实验证实miR-9-5p 与MEG3 或SOCS5 存在靶向关系。MEG3 和SOCS5 显著抑制宫颈癌细胞的增殖、迁移与侵袭能力（均P<0.01）,miR-9-5p 显著提高细胞的增殖、迁移与侵袭能力（均P<0.01）。MEG3 和SOCS5 促进E-cadherin 表达、抑制vimentin表达;miR-9-5p 抑制E-cadherin 表达、促进vimentin 表达（P<0.05 或P<0.01）。结论: lncRNA MEG3 通过miR-9-5p/SOCS5 分子轴调控宫颈癌细胞的增殖、迁移、侵袭与EMT进程。