CircRRAS2 promotes myogenic differentiation of bovine MuSCs and is a novel regulatory molecule of muscle development.

The proliferation and myogenic differentiation of muscle stem cells (MuSCs) are important factors affecting muscle development and beef quality. There is increasing evidence that circRNAs can regulate myogenesis. We found a novel circRNA, named circRRAS2 that is significantly upregulated in the differentiation phase of bovine MuSCs. Here, we aimed to determine its roles in the proliferation and myogenic differentiation of these cells. The results showed that circRRAS2 was expressed in several bovine tissues. CircRRAS2 inhibited MuSCs proliferation and promoted myoblast differentiation. In addition, chromatin isolation by using RNA purification and mass spectrometry in differentiated muscle cells identified 52 RNA-binding proteins that could potentially bind to circRRAS2, in order to regulate their differentiation. The results suggest that circRRAS2 could be a specific regulator of myogenesis in bovine muscle.HighlightsCircRRAS2 expression is higher in DM cells than in GM cells.CircRRAS2 could significantly inhibit the proliferation and apoptosis of bovine MuSCs.CircRRAS2 promotes the differentiation of bovine MuSCs into myotubes.CircRRAS2 may exert regulatory effects through multiple RNA binding proteins.

CircUBE2Q2 promotes differentiation of cattle muscle stem cells and is a potential regulatory molecule of skeletal muscle development.

BACKGROUND: The growth and development of muscle stem cells (MuSCs) are significant events known to affect muscle plasticity, disease, meat production, and meat quality, which involves the types and functions of mRNA and non-coding RNA. Here, MuSCs were cultured from Guangxi fetal cattle. RNA sequencing was used to analyze the RNA expression of mRNA and non-coding RNAs during the cell proliferation and differentiation phases. RESULTS: Two thousand one hundred forty-eight mRNAs and 888 non-coding RNAs were differentially expressed between cell proliferation and differentiation phases, including 113 miRNAs, 662 lncRNAs, and 113 circRNAs. RT-qPCR verified the differential expression levels of mRNAs and non-coding RNAs, and the differentially expressed circUBE2Q2 was subsequently characterized. Expression profile analysis revealed that circUBE2Q2 was abundant in muscle tissues and intramuscular fat. The expression of cricUBE2Q2 was also significantly upregulated during MuSCs myogenic differentiation and SVFs adipogenic differentiation and decreased with age in cattle muscle tissue. Finally, the molecular mechanism of circUBE2Q2 regulating MuSCs function that affects skeletal muscle development was investigated. The results showed that circUBE2Q2 could serve as a sponge for miR-133a, significantly promoting differentiation and apoptosis of cultured MuSCs, and inhibiting proliferation of MuSCs. CONCLUSIONS: CircUBE2Q2 is associated with muscle growth and development and induces MuSCs myogenic differentiation through sponging miR-133a. This study will provide new clues for the mechanisms by which mRNAs and non-coding RNAs regulate skeletal muscle growth and development, affecting muscle quality and diseases.

Genome-wide analysis of long non-coding RNA in primary nasopharyngeal carcinoma by microarray.

AIMS: Alterations in the expression of several long non-coding RNAs (lncRNAs) have been found in primary nasopharyngeal carcinoma (NPC). However, the effect of lncRNA expression on primary NPC as well as the molecular mechanism of lncRNA remains vague. This study was to identify differentially expressed lncRNAs involved in NPC on a genome-wide scale and predict their potential functions. METHODS AND RESULTS: Using high-throughput microarray with 30,586 lncRNA and 26,109 mRNA probes, 856 lncRNAs and 767 mRNAs were expressed differentially between NPC and chronic nasopharyngitis tissues. Bioinformatic analysis (clustering analysis, gene ontology analysis and pathway analysis) was used for further research. Differentially expressed lncRNAs were subgrouped into three types and differentially expressed mRNAs were clustered into 28 pathways. The first coexpression network analysis revealed that 46 lncRNAs interacting with three mRNAs involved the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signalling pathway. Quantitative real-time polymerase chain reaction (PCR) verified 11 up- and down-regulated lncRNAs and eight mRNAs in NPC. The second coexpression network analysis showed that 23 significantly aberrantly expressed mRNAs interacted with three validated lncRNAs. CONCLUSIONS: This study could provide new insight into the molecular mechanisms of lncRNAs and their potential role in NPC for further study. These differentially expressed lncRNAs may act as novel biomarkers and therapeutic targets for NPC.

Aberrant expression levels of MTA1 and RECK in nasopharyngeal carcinoma: association with metastasis, recurrence, and prognosis.

OBJECTIVES: We investigated the expression and clinical value of MTA1 and RECK genes in patients with nasopharyngeal carcinoma (NPC). METHODS: We examined MTA1 and RECK expression in nasopharyngeal tissue from patients with chronic nasopharyngitis, lymph nodes with metastasis of NPC, and primary NPC tumor tissue by means of in situ hybridization and analyzed their correlation with the clinicopathologic features of NPC. RESULTS: The positive expression of MTA1 in the NPC tissues and metastatic lymph nodes was significantly higher than that in the chronic nasopharyngitis tissues (p < 0.05). The positive expression of RECK in the NPC tissues and metastatic lymph nodes was significantly lower than that in the chronic nasopharyngitis tissues (p < 0.05). The RECK expression level was inversely correlated with the MTA1 expression level in the NPC tissues (p < 0.05). The increased MTA1 and decreased RECK expressions in the NPC tissues had no association with gender, age, T-stage, or clinical stage (p > 0.05). However, they had a positive correlation with cervical lymph node metastasis, tumor recurrence, and 5-year overall survival rate of the patients with NPC (p < 0.05). Moreover, multivariate analysis showed that MTA1 and RECK expressions were independent prognostic factors for survival (p < 0.05). CONCLUSIONS: The conversely abnormal expression levels of MTA1 and RECK may be collectively involved in progression of malignancies and may serve as molecular predictors for metastasis, recurrence, and prognosis of NPC.

Allelic imbalance and abnormal expression of FHIT in endemic nasopharyngeal carcinoma: association with clinicopathological features.

The FHIT gene is involved in the pathogenesis of many cancers. The aim of this study was to investigate allelic imbalance (AI) pattern at FHIT locus and alteration of FHIT gene in nasopharyngeal carcinoma (NPC) and analyzed potential correlation between AI, FHIT mRNA expression and clinicopathological factors. We examined AI, including loss of heterozygosity (LOH) and microsatellite instability (MSI), at FHIT locus in 41 cases of NPC by microsatellite analysis and FHIT gene status in 30 cases of NPC by nested reverse transcriptase-polymerase chain reaction and DNA sequencing. The frequencies of LOH and MSI at FHIT locus in NPC were 70.7% (29/41) and 36.6% (15/41), respectively. Thirteen of thirty (43.3%) NPCs exhibited aberrant FHIT transcripts. LOH and abnormal FHIT expression were correlated with advanced clinical stage and higher titers of immunoglobulin (Ig) A against Epstein-Barr virus capsid antigen (EBVCA-IgA) (p < 0.05). Abnormal FHIT expression was also correlated with tumor recurrence (p < 0.05). MSI was correlated with early clinical stage and higher titers of EBVCA-IgA (p < 0.05). AI at FHIT locus is a common event and contributes to genetic imbalance in NPC. The abnormalities of FHIT, presumably associated with genetic imbalance at FHIT locus, might be involved in the development and the tumor recurrence of NPC.

CircRNAs: potency of protein translation and feasibility of novel biomarkers and therapeutic targets for head and neck cancers.

Circular RNAs (circRNAs), a new star noncoding RNA (ncRNA), show stability, conservation, abundance, and tissue and stage specificity. They act as key regulators of biological processes. They target the mRNAs of many other different genes or signaling pathways, and closely link associated genes into regulatory networks. Growing evidence has demonstrated that circRNAs may play an important role in the carcinogenesis, progression and chemoradiation resistance of many cancers including head and neck cancers (HNC). CircRNA, like other ncRNA, such as miRNA, lncRNA, usually is considered to be non-protein coding transcript. However, recent studies indicated that abnormal translation of circRNAs may be involved in human diseases. In this review, we collected the origin, classification, characteristics, function of circRNAs, exosmal circRNAs, and then synthesize current study results to highlight aberration of circRNAs in various types of HNC, and try to clarify the molecular mechanisms of circRNAs affecting the pathogenesis and progression of HNC, as well as pay particular attention to provide a new avenue to the diagnosis and treatment strategy for HNC.

Integrated analysis of transcriptome profiling predicts potential lncRNA and circRNA targets in human nasopharyngeal carcinoma.

Non-coding RNAs (ncRNAs) regulate numerous genes and influence the progression of various human diseases, including cancer. The role of regulatory ncRNAs implicated in nasopharyngeal carcinoma (NPC), as well as their target genes, remains unclear. The present study aimed to investigate specific long non-coding (lnc)RNAs, circular RNAs (circRNAs) and mRNAs associated with the molecular pathogenesis of NPC, and to predict the underlying target genes of specific lncRNAs and circRNAs. The expression levels of lncRNAs, circRNAs and mRNAs in NPC and chronic nasopharyngitis tissues were detected and analyzed using microarray and bioinformatics techniques. A total of 2.80% lncRNAs (425 upregulated and 431 downregulated) were significantly differentially expressed (DE) between the two tissue types. Additionally, 0.96% circRNAs (18 upregulated and 13 downregulated) were significantly DE, while 2.94% mRNAs (426 upregulated and 341 downregulated) were significantly DE between the two tissue types. In total, 420 NPC-associated nearby encoding genes (196 up- and 224 downregulated) of the DE lncRNAs were identified. Overlap analysis identified 23 DE circRNAs and their corresponding target genes, with 37 microRNAs and 50 mRNAs, from which 14 interaction networks were constructed. Subsequent pathway analysis revealed 221 DE target genes corresponding to 31 key signaling pathways associated with NPC, 14 of which may represent hub genes associated with NPC pathophysiology. Thus, certain lncRNAs, circRNAs and mRNAs are aberrantly expressed in NPC tissues, and partially specific lncRNAs, circRNAs and their target genes may influence the tumorigenesis and progression of NPC. Target prediction and regulatory network identification may help to determine the pathogenic mechanisms of NPC.

Head and neck rhabdomyosarcoma: follow-up results of four cases and review of the literature.

Head and neck rhabdomyosarcoma (HNRMS) is exceedingly rare and poorly documented. The difficult diagnosis often causes a poor prognosis and high mortality. Hence, we report 4 cases of HNRMS and their follow-up outcomes, and review the clinicopathological features of this rare tumor. The 4 patients ranged in age from 5 to 29 years. Among them, 3 patients had a good prognosis after combination of radiotherapy and chemotherapy or surgery alone. Another patient survived for only 3 months after diagnosis without therapy. Deeply insight into HNRMS might improve the ability of diagnosis and treatment for this disease.

Analytical and clinical evaluation of CYFRA 21-1 by electrochemiluminescent immunoassay in head and neck squamous cell carcinoma.

This paper attempts to evaluate the clinical usefulness of CYFRA 21-1 as a serum tumour marker in patients with head and neck squamous cell carcinoma (HNSCC). The serum concentration of CYFRA 21-1 was measured utilizing a new electrochemiluminescent immunoassay (ECLIA) in 142 patients with HNSCC before and after treatment, 68 patients with benign tumours of the head and neck, and 50 healthy controls. Serum levels of CYFRA 21-1 in patients with HNSCC were significantly higher than those of benign tumours and healthy controls (p < 0.001). The diagnostic sensitivity and specificity of CYFRA 21-1 for HNSCC were 62 per cent and 100 per cent, respectively. The positive rates of CYFRA 21-1 increased with progression of HNSCC, serum CYFRA 21-1 levels were related to the tumour stage expressed by primary tumour (T) and nodal status (N) (p < 0.001), but not related to patient age, gender, smoking and drinking habit, or histopathological grade (p > 0.05). Post-treatment levels of CYFRA 21-1 in HNSCC decreased significantly (p < 0.001). Among 38 patients with clinical or radiological evidence of a recurrence during follow-up, 78.9 per cent (30 of 38) showed an increase in CYFRA 21-1. The analytical ECLIA performance for serum CYFRA 21-1 provides a new means of clinical assessment for HNSCC. The results of ECLIA suggest that the serum marker CYFRA 21-1 is valuable not only for diagnosis but also for close monitoring of patients with HNSCC.

Long non-coding RNAs as novel biomarkers and therapeutic targets in head and neck cancers.

Long non-coding RNAs (lncRNAs) are generally defined as RNA molecules greater than 200 nt in length and without protein-coding property that different from housekeeping RNAs such as tRNAs, rRNAs, and snRNAs, and independent from small RNAs with specific molecular processing machinery such as micro- or piwi-RNAs. LncRNAs are a novel class of mRNA-like transcripts which contribute to cancer development and progression and accelerate cancer cells proliferation, invasion, metastasis, and apoptosis. These research results indicate the potential of lncRNAs as prospective novel biomarkers for diagnosis, therapeutic targets and prognosis for cancers. In this review, we synthesize present study results to highlight aberration of lncRNAs in various types of head and neck cancers, and try to clarify the molecular mechanisms of lncRNAs affecting the oncogenesis and progression of head and neck cancer, as well as pay particular attention to provide a new avenue to the diagnosis and treatment strategy.

Aberrant SATB1 expression is associated with Epstein-Barr virus infection, metastasis and survival in human nasopharyngeal cells and endemic nasopharyngeal carcinoma.

Special AT-rich sequence-binding protein 1 (SATB1) has been identified as a key factor in the progression of some cancers, functioning as a global genome organizer and chromatin regulator. We examined the levels of SATB1 mRNA expression in NPC cell lines 5-8F (high metastasis) and 6-10B (low metastasis) and immortalized human nasopharyngeal epithelial cells NP69-SV40T by quantitative real-time PCR. We also examined the protein expression levels of SATB1 in 72 cases of nasopharyngeal carcinoma (NPC) tissues and 30 cases of normal nasopharyngeal (NNP) tissues by immunohistochemistry, and then assessed the correlations between SATB1 expression and clinicopathological factors. The expression level of SATB1 mRNA in 5-8F was much higher than those in 6-10B and NP69-SV40T (P<0.05). The expression level of SATB1 mRNA in 6-10B was higher than in NP69-SV40T, but the difference was not statistically significant (P>0.05). The positive expression rates of SATB1 protein in NPC (38/72, 52.8%) were significantly higher than in NNP (4/30, 13.3%) (P<0.05). SATB1 protein levels in NPC were not associated with gender, age, and T stage (P>0.05), but positively correlated with the titers of EBVCA-IgA, metastasis (N and M stage), recurrence, and survival (P<0.05). Multivariate analysis showed that the overexpression of SATB1 protein is an independent prognostic factor for NPC. The expression levels of SATB1 were obviously upregulated in primary NPC tissues and human NPC cell lines. Therefore, SATB1 may be a valuable predictor in assessing the metastasis, recurrence, and prognosis of NPC.

Silencing SATB1 influences cell invasion, migration, proliferation, and drug resistance in nasopharyngeal carcinoma.

Special AT rich sequence binding protein 1 (SATB1) play an important role in many cancers, but the role of SATB1 in nasopharyngeal carcinoma (NPC) is still not full understand. Immunofluorescence staining showed that SATB1 was mainly localized in the nuclei in CNE-2 cell. After successful down-regulation of SABT1 in NPC cell line CNE-2 by shRNA, compared to parental CNE-2 and control shRNA group, the capacity of the proliferation, migration, invasion and drug resistance of CNE-2 cell was reduced, which indicated that SATB1 may be involved in NPC development and progression. SATB1 may be a promising therapeutic target for nasopharyngeal carcinoma.

Concurrent alterations of RAGE, RECK, and MMP9 protein expression are relevant to Epstein-Barr virus infection, metastasis, and survival in nasopharyngeal carcinoma.

This study aimed to concurrently investigate the expressions of receptor for advanced glycation end products (RAGE), reversion inducing cysteine-rich protein with Kazal motifs (RECK) and matrix metalloproteinase 9 (MMP9) in nasopharyngeal carcinoma (NPC) and their correlations with clinicopathological properties. Using immunohistochemistry, we found that RECK expression was downregulated in NPC tissues compared with chronic nasopharyngitis (CNT) tissues, while RAGE and MMP9 expressions were upregulated. We further found that RECK expression level was inversely correlated with MMP9 expression level in NPC, whereas RAGE expression level was positively correlated with MMP9 expression level. Moreover, aberrant expressions of these proteins had a positive correlation with the titers of EBVCA-IgA, lymphatic metastasis, recurrence and survival. Together, these findings suggest that dysregulations of RECK and RAGE expressions may be collectively involved in tumor progression of NPC by regulating MMP9 expression and that they may be a good prognostic predictors for NPC.

[Cloning and tissue expression analysis of creatine kinase (M-CK) cDNA from the mandarin fish, Siniperca chuatsi].

The creatine kinase (CK) cDNA from the mandarin fish Siniperca chuatsi was cloned by RT-PCR and rapid amplification of cDNA ends (RACE) methods. The structural characteristics and phylogeny of this gene were analyzed. Sequence analysis revealed a 1586 bp cDNA sequence containing 92 bp 5'-untranslated region, 348 bp 3'-untranslated region and 1146 bp open reading frame (ORF), which encoded 381 amino acids. Conserved sequence blocks of vertebrate CKs and diagnostic boxes for the muscle CK (M-CK) isozyme were identified in S. chuatsi CK. Siniperca chuatsi CK showed a higher similarity with vertebrates M-CK isozyme than other CK isozymes (Brain CK, Mitochondrial CKs) and grouped with M-CK isozyme in CK phylogeny, which strongly supported that S. chuatsi CK belongs to M-CK isozyme type. Semi-quantitative RT-PCR analysis demonstrated that the M-CK transcript expression varied among the different tissues and was detected at a high level in skin, ovary, kidney, stomach, muscle and heart, but lower in eye, brain and liver.