超级增强子长链非编码RNA LINC01232在大肠癌组织中的表达及其对癌细胞生物学行为的影响

目的 探讨超级增强子长链非编码RNA(SE-LncRNA)LINC01232在大肠癌组织中的表达水平及其对大肠癌细胞生物学行为的影响.方法 通过基因芯片筛选4对大肠癌患者组织(癌组织和癌旁组织)中差异表达的SE-LncRNA,发现LINC01232在癌组织中显著高表达(Fold Change=2.6569,P=0.02...     

Expression and clinical significance of PTPN12 in clear cell renal cell carcinoma

BackgroundClear cell renal cell carcinoma (ccRCC) is a common urological disease. Expression of the protein tyrosine phosphatase 12 gene (PTPN12) is decreased in many cancers; however, the relationship between PTPN12 gene function and renal cancer remains unclear.MethodsWe detected PTPN12 protein expression in ccRCC and corresponding normal tissues from 64 patients with ccRCC by immunohistochemistry, and relative PTPN12 mRNA levels by real-time quantitative polymerase chain reaction. The relationships between the relative expression levels of PTPN12 mRNA and the patients’ clinical data were analyzed.ResultsPTPN12 protein and mRNA expression levels were significantly lower in ccRCC compared with the corresponding normal tissues. The mRNA expression levels in the ccRCC and corresponding normal tissues from the 64 patients with ccRCC were 0.459±0.445 and 1.001±0.128, respectively, compared with the control (glyceraldehyde 3-phosphate dehydrogenase). There was a significant correlation between relative expression of PTPN12 mRNA in ccRCC tissues and tumor diameter and clinical stage.ConclusionThe expression levels of PTPN12 protein and mRNA were significantly lower in ccRCC tissues compared with normal tissues. The role of PTPN12 may provide new insights and evidence to aid the diagnosis and targeted therapy of ccRCC.     

Long noncoding RNA LINC01296 plays an oncogenic role in colorectal cancer by suppressing p15 expression

ObjectiveTo examine the role of the long noncoding RNA LINC01296 in colorectal carcinoma (CRC) and to explore the underlying mechanism.MethodsWe detected LINC01296 expression levels in a cohort of 51 paired CRC and normal tissues. We also assessed the effects of LINC01296 on cell proliferation and apoptosis in CRC cells in vitro, and measured its effect on tumor growth in an in vivo mouse model. We identified the potential downstream targets of LINC01296 and assessed its regulatory effects.ResultsExpression levels of LINC01296 were elevated in 37/51 CRC tissues compared with the corresponding normal tissues and were significantly associated with tumor stage, lymph node metastasis, and distant metastasis. Knockdown of LINC01296 using antisense oligonucleotides inhibited cell proliferation and promoted apoptosis of colon cancer cells in vitro and inhibited tumor growth in vivo. Knockdown of LINC01296 also significantly increased the gene expression of p15 in colon cancer cells. LINC01296-specific suppression of p15 was validated by the interaction between enhancer of zeste homolog 2 and LINC01296.ConclusionOverexpression of LINC01296 suppressed the expression of p15 leading to CRC carcinogenesis. These findings may provide the basis for novel future CRC-targeted therapies.     

LINC00115 regulates lung adenocarcinoma progression via sponging miR-154-3p to modulate Sp3 expression

The most commonly diagnosed and most lethal subtype of lung cancer is lung adenocarcinoma (LUAD). Therefore, more detailed understanding of the potential mechanism and identification of potential targets of lung adenocarcinoma is needed. A growing number of reports reveals that long non-coding RNAs (lncRNAs) play crucial roles in cancer progression. In present study, we found that lncRNA LINC00115 was upregulated in LUAD tissues and cells. Functional studies revealed that LINC00115 knockdown inhibits the proliferation, growth, invasion, and migration of LUAD cells. Mechanically, we indicated that miR-154-3p is target microRNA of LINC00115, and the effect of downregulated LINC00115 on LUAD cells was partially reversed by the miR-154-3p antisense oligonucleotide (ASO-miR-154-3p). Further investigation revealed that Specificity protein 3 (Sp3) directly interacted with miR-154-3p, and the Sp3 level was positively correlated with the LINC00115 expression. Rescue experiments further showed that Sp3 overexpression partially restored the effect of downregulated LINC00115 on LUAD cells. Similarly, in vivo experiments confirmed that downregulated LINC00115 inhibited xenograft growth and Sp3 expression. Our results demonstrated that LINC00115 knockdown inhibited LUAD progression via sponging miR-154-3p to modulate Sp3 expression. These data indicate that the LINC00115/miR-154-3p/Sp3 axis can be a potential therapeutic target of LUAD.     

Hsa_circ_0042823 accelerates cancer progression via miR-877-5p/FOXM1 axis in laryngeal squamous cell carcinoma

BackgroundLaryngeal squamous cell carcinoma (LSCC) is a common malignant tumour of the head and neck. Our previous study reveals that the circular RNA (circRNA) hsa_circ_0042823 is abnormally expressed in LSCC, suggesting that hsa_circ_0042823 is closely associated with LSCC. Here, we attempted to explore the molecular mechanism of hsa_circ_0042823 in LSCC.MethodsQuantitative real-time PCR and western blot were performed to assess the expression of gene and protein in human laryngeal carcinoma cells, TU212 and TU686. MTT and transwell assays were performed to examine cell proliferation, migration and invasion. The relationship among hsa_circ_0042823, miR-877-5p and forkhead box M1 (FOXM1) was verified by luciferase reporter assay. Finally, we constructed a subcutaneous tumour mouse model to analyse in vivo growth of LSCC cells following knockdown of hsa_circ_0042823.ResultsCompared with normal human bronchial epithelial cells (HBECs), hsa_circ_0042823 was highly expressed in the LSCC cell lines (AMC-HN-8 and TU686). Further studies demonstrated that hsa_circ_0042823 interacted with miR-877-5p, and FOXM1 was the target of miR-877-5p. Hsa_circ_0042823 promoted the expression of FOXM1 via its ceRNA activity on miR-877-5p. Hsa_circ_0042823 overexpression promoted proliferation, migration and invasion of AMC-HN-8 cells through regulating miR-877-5p/FOXM1 axis. Additionally, inhibition of hsa_circ_0042823 inhibited growth of LSCC in vivo via miR-877-5p/FOXM1 axis.ConclusionsHsa_circ_0042823/miR-877-5p/FOXM1 axis participates in the progression of LSCC. This work demonstrates that hsa_circ_0042823 accelerates cancer progression by regulating miR-877-5p/FOXM1 axis in LSCC. Therefore, this study may provide new insights into the pathogenesis of LSCC.

KEY MESSAGES

• Hsa_circ_0042823 promotes FOXM1 expression by sponging miR-877-5p.
• Hsa_circ_0042823 promotes proliferation, migration, invasion of LSCC cells.
• Hsa_circ_0042823 knockdown inhibits tumour growth of LSCC via miR-877-5p/FOXM1 axis.

     

miR-301-3p directly regulates Cx43 to mediate the development of gastric cancer

ObjectiveIdentifying novel biomarkers involved in the development of gastric cancer (GC) can provide potential therapeutic strategies and improve clinical prognosis. miR-301-3p and Cx43 are reportedly dysregulated in GC. miR-301-3p and Cx43 interaction, and their functions in GC progression, are still poorly understood.MethodsThe expression levels of miR-301-3p and Cx43 in GC tissues and cell lines with various differentiation degrees were evaluated by RT-qPCR. The interaction between miR-301-3p and Cx43 was assessed by dual-luciferase reporter assays. CCK8 and Transwell assays were employed to assess the effects of the miR-301-3p-Cx43 axis on GC cell proliferation, migration, and invasion.ResultsCx43 was significantly downregulated in GC tissues and cell lines, while miR-301-3p expression was negatively correlated with Cx43 mRNA levels. The expression levels of Cx43 and miR-301-3p were closely associated with the differentiation, TNM stage, vascular invasion, and lymph node metastasis status of GC patients. Cx43 overexpression could suppress the proliferation, migration, and invasion of GC cells. Cx43 mRNA is a direct target of miR-301-3p, and transfection of an miR-301-3p mimic could reverse the inhibitory effects of Cx43.ConclusionThe miR-301-3p-Cx43 axis is involved in the development and progression of GC by affecting the proliferation, migration, and invasion of GC cells.     

LINC00261和miR-522-3p在结直肠癌中的表达及其与患者预后的关系

目的检测长链非编码RNA LINC00261、微小RNA-522-3p(miR-522-3p)在结直肠癌组织中的表达水平,并探讨两者的相关性及其与患者预后的关系。方法收集2013年1月至2014年12月在该院行手术切除治疗的68例结直肠癌组织及其配对的癌旁组织,应用实时荧光定量PCR检测结直肠癌和癌旁组织中LINC00261、miR-522-3p的表达水平;分析LINC00261、miR-522-3p的表达与结直肠癌患者临床病理参数的关系;Kaplan-Meier生存模型分析LINC00261、miR-522-3p表达对结直肠癌患者生存率的影响;多因素Cox回归分析影响结直肠癌患者预后的独立危险因素;Pearson相关分析检测LINC00261与miR-522-3p在结直肠癌组织中表达水平的相关性。结果LINC00261在结直肠癌组织中表达水平低于其在癌旁组织中的表达水平(P<0.05),miR-522-3p在结直肠癌组织中表达水平高于其在癌旁组织中的表达水平(P<0.05)。LINC00261在结直肠癌组织中的表达水平与分化程度、临床分期和淋巴结转移情况相关(P<0.05),miR-522-3p在结直肠癌组织中的表达水平与临床分期和淋巴结转移情况相关(P<0.05)。LINC00261低表达组的预后比LINC00261高表达组预后更差(P<0.05),miR-522-3p高表达组的预后比miR-522-3p低表达组的预后更差(P<0.05)。高临床分期、低分化程度、淋巴结转移、LINC00261低表达和miR-522-3p高表达是结直肠癌患者预后不良的独立危险因素(P<0.05)。结直肠癌组织中LINC00261与miR-522-3p的表达呈负相关(P<0.05)。结论结直肠癌组织中LINC00261表达下调、miR-522-3p表达上调,二者表达呈负相关,且均与结直肠癌患者的不良病理参数和预后相关。LINC00261、miR-522-3p可能是结直肠癌潜在的预后标志物和治疗靶标。     

Novel immune‐related signature based on immune cells for predicting prognosis and immunotherapy response in clear cell renal cell carcinoma

BackgroundClear cell renal cell carcinoma (ccRCC) is the most common malignant tumor of the kidney and is characterized by poor prognosis. We sought to build an immune‐related prognostic signature and investigate its relationship with immunotherapy response in ccRCC.MethodsImmune‐related genes were identified by ssGSEA and WGCNA. The prognostic signature was conducted via univariate, least absolute shrinkage and selection operator, and multivariable Cox regression analyses. Kaplan‐Meier analysis, PCA, t‐SNE, and ROC were used to evaluate the risk model.ResultsA total of 119 immune‐related genes associated with prognosis were screened out. Six immune‐related genes (CSF1, CD5L, AIM2, TIMP3, IRF6, and HHLA2) were applied to construct a prognostic signature for KIRC. Kaplan–Meier analysis showed that patients in high‐risk group had a poorer survival outcome than in low‐risk group. The 1‐, 3‐ and 5‐year AUC of the prognostic signature was 0.754, 0.715, and 0.739, respectively. Univariate and multivariate Cox regression models demonstrated that the risk signature was an independent prognostic factor for KIRC survival. GSEA analysis suggested that the high‐risk group was concentrated on immune‐related pathways. The high‐risk group with more regulatory T‐cell infiltration showed a higher expression of immune negative regulation genes. The risk score had positively relationship with TIDE score and negatively with the response of immunotherapy. The IC50 values of axitinib, sunitinib, sorafenib, and temsirolimus were lower in the high‐risk group.ConclusionOur study defined a robust signature that may be promising for predicting clinical outcomes and immunotherapy and targeted therapy response in ccRCC patients.     

MiR-454 promotes the progression of human non-small cell lung cancer and directly targets PTEN

PurposeMicroRNA-454 has been proven dysregulated in some human malignancies and correlated with tumor progression. However, its expression and function in non-small cell lung cancer (NSCLC) is still unclear. Thus, the aim of this study was to explore the effects of miR-454 in NSCLC tumorigenesis and development.MethodsUsing quantitative RT-PCR, we detected miR-454 expression in NSCLC cell lines and primary tumor tissues. The association of miR-454 expression with clinicopathological factors and prognosis was also analyzed. Then, the effects of miR-454 on the biological behavior of NSCLC cells were investigated. At last, the potential regulatory function of miR-454 on PTEN expression was confirmed.ResultsmiR-454 was found to be up-regulated in NSCLC tissues and cell lines. High miR-454 expression was closely correlated with lymph node metastasis, advanced TNM stage, and shorter overall survival. Multivariate regression analysis corroborated that miR-454 overexpression was an independent unfavourable prognostic factor for patients with NSCLC. Down-regulation of miR-454 could significantly reduce NSCLC cell proliferation, enhance cell apoptosis, and impair cell invasion and migration in vitro, while up-regulation of miR-454 showed opposite effects. Further, PTEN was confirmed as a direct target of miR-454 by using Luciferase Reporter Assay.ConclusionsThese findings indicate that miR-454 may act as an oncogene in NSCLC and would serve as a potential therapy target for this disease.     

Circular RNA circ_0010235 sponges miR-338-3p to play oncogenic role in proliferation,migration and invasion of non-small-cell lung cancer cells through modulating KIF2A

BackgroundCircular RNA microarray analysis showed hsa_circ_0010235 (circ_0010235) was highly upregulated in non-small-cell lung cancer (NSCLC) patients; however, its role in carcinogenesis and development of NSCLC cells was unrevealed. Here, we intended to investigate role and mechanism of circ_0010235 in NSCLC proliferation, migration and invasion.Methods and resultsExpression of circ_0010235, microRNA (miR)-338-3p and kinesin family member 2A (KIF2A) was detected by quantitative real-time PCR, western blotting and immunohistochemistry (IHC). Cell progression was measured by cell-counting kit-8 assay, 5-ethynyl-2-deoxyuridine (EdU) assay, flow cytometry, transwell assay, western blotting, IHC and xenograft experiment. The relationship among circ_0010235, miR-338-3p and KIF2A was determined by dual-luciferase reporter assay, RNA immunoprecipitation and Pearson’s correlation analysis. Expression of circ_0010235 was increased in human NSCLC tissues and cells, accompanied with miR-338-3p downregulation and KIF2A upregulation. Essentially, circ_0010235 could sponge miR-338-3p via target binding, and miR-338-3p downstream targeted KIF2A. Functionally, exhaustion of circ_0010235 induced apoptosis rate of NSCLC cells and curbed cell viability, EdU incorporation, migration rate and invasion rate, accompanied with higher E-cadherin and lower N-cadherin expression. Additionally, re-expression of miR-338-3p prompted above similar effects in NSCLC cells in vitro. Contrarily, miR-338-3p blockage partially counteract the effects of circ_0010235 exhaustion; plus, restoration of KIF2A could attenuate miR-338-3p role, as well. Notably, interfering circ_0010235 delayed tumour growth of NSCLC cells by promoting miR-338-3p and E-cadherin expression, and depressing KIF2A, ki-67 and N-cadherin expression.Conclusionscirc_0010235 could be a novel identified oncogenic circRNA in NSCLC, and targeting miR-338-3p/KIF2A axis was one regulatory mechanism underlying circ_0010235.

KEY MESSAGE

• Circ_0010235 was an upregulated circRNA in NSCLC patients and cells.
• Interfering circ_0010235 restrained NSCLC cell proliferation and metastasis in vitro and in vivo.
• miR-338-3p per se suppressed NSCLC in vitro and its downregulation diminished the tumour-suppressive role of circ_0010235 blockage in NSCLC cells.
• miR-338-3p could downstream target KIF2A and be sponged by circ_0010235.

     

Prognostic urinary miRNAs for the assessment of small renal masses

BackgroundRenal cell carcinoma (RCC) is often detected incidentally as a small renal mass (SRM; pT1a, ≤4 cm). It is clinically challenging to predict progression in patients with SRMs. This is largely due to the recent recognition of clinically progressive and non-progressive RCC-SRMs. It is critical to accurately stratify SRM patients according to risk to avoid unnecessary treatment. This is especially significant for elderly and infirm patients, where the risk of surgery outweighs mortality from SRMs.MethodsWe employed a qRT-PCR array-based approach and targeted qRT-PCR to identify and validate early, non-invasive diagnostic and prognostic biomarkers of RCC-SRMs. In total, we evaluated eighty urine samples, including 30 renal oncocytoma (≤4 cm) cases, 26 progressive and 24 non-progressive clear cell RCC-SRM (ccRCC-SRM) cases.ResultsWe identified nine urinary miRNAs which displayed significantly elevated expression in ccRCC-SRMs (pT1a; ≤4 cm) relative to renal oncocytoma (≤4 cm). Additionally, miR-328-3p displayed significantly down-regulated expression in progressive relative to non-progressive ccRCC-SRMs. Patients with elevated miR-328-3p expression had significantly longer overall survival (HR = 0.29, 95% CI = 0.08–1.03, p = 0.042) compared to patients with low miR-328-3p expression. We also found no significant association between miR-328-3p expression levels and gender, age, laterality, tumor size, or grade, suggesting that miR-328-3p is an independent prognostic biomarker.ConclusionsOur in-depth miRNA profiling approach identified novel biomarkers for early-stage ccRCC-SRMs. Pretreatment characterization of urinary miRNAs may provide insight into early RCC progression and could potentially aid clinical decision-making, improving patient management and reducing overtreatment.     

肾透明细胞癌患者血清细胞外囊泡中miR-99b-3p水平检测的临床价值

目的 探讨肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)患者血清细胞外囊泡(extracellular vesicles,EVs)中miR-99b-3p的水平变化,评估其作为ccRCC非侵入性辅助诊断分子标志物的临床价值.方法 收集2016年12月至2018年10月东部战区总...     

miR-486-5p inhibits cell growth of papillary thyroid carcinoma by targeting fibrillin-1

Previous studies have demonstrated that miR-486-5p functions as a tumor suppressor or oncogene in various types of cancer. In the present study, we showed that miR-486-5p was significantly down-regulated in papillary thyroid carcinoma (PTC) tissues and cell lines, whereas miR-486-5p down-regulation inhibited PTC cell proliferation and increased apoptosis. Conversely, under-expression of miR-486-5p enhanced PTC cell proliferation and decreased apoptosis. Fibrillin-1 (FBN1) was shown to be a direct target of miR-486-5p and inversely regulated by miR-486-5p. FBN1 silencing led to decreased PTC cell proliferation and enhanced apoptosis in vitro, similar to that mediated by miR-486-5p. Furthermore, miR-486-5p over-expression or FBN1 knock-down inhibited, while up-regulation of FBN1 boosted xenograft tumor formation in vivo. Our data suggest that miR-486-5p induces PTC cell growth inhibition and apoptosis by targeting and suppressing FBN1. Thus, miR-486-5p/FBN1 might provide a promising therapeutic target for PTC treatment.     

CircHIPK3 regulates melanoma cell behaviors by binding with miR-215–5p to upregulate YY1

ObjectTo investigate the role of circHIPK3 in melanoma.MethodsBioinformatics analysis and experiments including RT-qPCR, Pearson's correlation analysis, luciferase reporter, Western blot, and RIP assays were applied to explore the function and mechanism of circHIPK3 in melanoma.ResultsCircHIPK3 expression was strikingly upregulated while miR-215–5p was downregulated in melanoma tissues and cell lines. Pearson's correlation analysis unveiled circHIPK3 expression was positively correlated with Ki-67 (a marker of proliferation), which implied that circHIPK3 may play a vital role in the progression of melanoma. In mechanism, luciferase reporter and RIP assays validated that circHIPK3 was able to bind with miR-215–5p. Moreover, we confirmed that overexpression of circHIPK3 could facilitate cell proliferation and depress cell apoptosis in melanoma while overexpression of miR-215–5p exerted opposite effects. Besides, our findings indicated that miR-215–5p overexpression significantly reversed the circHIPK3 overexpressing-mediated promotive effect on cell proliferation and inhibitory effect on cell apoptosis. Furthermore, we found that miR-215–5p could directly target YY1. Upregulation of YY1 could notably offset the inhibitory effect of circHIPK3 downregulation on cell proliferation and the promotive effect on cell apoptosis.ConclusionOur study corroborated that circHIPK3 regulated melanoma cell behaviors via the miR-215–5p/YY1 axis, which might provide a novel insight for the treatment of melanoma patients.