Five genes involved in circular RNA-associated competitive endogenous RNA network correlates with metastasis in papillary thyroid carcinoma.

This study aimed to identify potential circular RNA (circRNA), microRNA (miRNA) and mRNA biomarkers as well as their underlying regulatory mechanisms in papillary thyroid carcinoma (PTC). Three microarray datasets from the Gene Expression Omnibus database as well as expression data and clinical phenotype from The Cancer Genome Atlas (TCGA) were downloaded, followed by differential expression, functional enrichment, protein-protein interaction (PPI), and module analyses. The support vector machine (SVM)-recursive feature elimination (RFE) algorithm was used to screen the key circRNAs. Finally, the mRNA-miRNA-circRNA regulatory network and competitive endogenous RNA (ceRNA) network were constructed. The prognostic value and clinical correlations of key mRNAs were investigated using TCGA dataset, and their expression was validated using the UALCAN database. A total of 1039 mRNAs, 18 miRNAs and 137 circRNAs were differentially expressed in patients with PTC. A total of 37 key circRNAs were obtained using the SVM-RFE algorithm, whereas 46 key mRNAs were obtained from significant modules in the PPI network. A total of 11 circRNA-miRNA pairs and 40 miRNA-mRNA pairs were predicted. Based on these interaction pairs, 46 circRNA-miRNA-mRNA regulatory pairs were integrated, of which 8 regulatory pairs in line with the ceRNA hypothesis were obtained, including two circRNAs (circ_0004053 and circ_0028198), three miRNAs (miR-199a-5p, miR-199b-5p, and miR-7-5p), and five mRNAs, namely APOA2, CCL20, LPAR5, MFGE8, and TIMP1. Survival analysis showed that LPAR5 expression was associated with patient survival. APOA2 expression showed significant differences between metastatic and non-metastatic tumors, whereas CCL20, LPAR5, MFGE8 and TIMP1 showed significant differences between metastatic and non-metastatic lymph nodes. Overall, we identified several potential targets and regulatory mechanisms involved in PTC. APOA2, CCL20, LPAR5, MFGE8, and TIMP1 may be correlated with PTC metastasis.

Protective effects of dexmedetomidine on cerebral ischemia/reperfusion injury via the microRNA-214/ROCK1/NF-κB axis.

BACKGROUND: Cerebral ischemia/reperfusion injury (CIRI) is a complication of surgical procedure associated with high mortality. The protective effect of dexmedetomidine (DEX) on CIRI has been explored in previous works, yet the underlying molecular mechanism remains unclear. Our study explored the protective effect of DEX and its regulatory mechanism on CIRI. METHODS: A CIRI rat model was established using middle cerebral artery occlusion (MCAO). Neurological deficit scores for rats received MCAO modeling or DEX treatment were measured. Cerebral infarction area of rats was detected by TTC staining, while damage of neurons in hippocampal regions of rats was determined by hematoxylin-eosin (HE) staining. Apoptosis rate of neurons in hippocampal regions was examined by TUNEL staining. The dual-luciferase assay was performed to detect the binding of microRNA-214 (miR-214) to Rho-associated kinase 1 (ROCK1). RESULTS: DEX treatment significantly reduced infarction area of MCAO rats and elevated miR-214 expression. Injection of miR-214 inhibitor attenuated the effect of DEX in MCAO rats by increasing the area of cerebral infarction in rats and apoptosis rate of hippocampal neurons. ROCK1 was targeted and negatively regulated by miR-214. The overexpression of ROCK1 led to activation of NF-κB to aggravate CIRI. CONCLUSION: Therapeutic effects of DEX on CIRI was elicited by overexpressing miR-214 and impairing ROCK1 expression and NF-κB activation. Our finding might provide novel insights into the molecular mechanism of DEX in rats with CIRI.

miRNA-520c-3p accelerates progression of nasopharyngeal carcinoma via targeting RAB22A.

[This retracts the article DOI: 10.3892/ol.2019.11144.].

lncRNA SNHG7 promotes tumorigenesis of nasopharyngeal carcinoma via epithelial-to-mesenchymal transition.

[This retracts the article DOI: 10.3892/ol.2020.11397.].

lncRNA SNHG7 promotes tumorigenesis of nasopharyngeal carcinoma via epithelial-to-mesenchymal transition.

Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors. Studies have indicated that long noncoding RNAs (lncRNAs) function as important regulators in progression of tumorigenesis. In this study, lncRNA small nucleolar RNA host gene 7 (SNHG7) was selected to identify how it functioned in the development of NPC. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to detect SNHG7 expression in paired NPC patient tissue samples and cell lines. The role of SNHG7 in the metastasis of NPC was detected through scratch wound assay and Transwell assay. RT-qPCR and western blot assay were used to discover the function of SNHG7 in epithelial-to-mesenchymal transition (EMT) process. Tumor metastasis assay was also performed in vivo. In this study, RT-qPCR results showed that SNHG7 expression in NPC samples was remarkably higher when compared with that in adjacent ones. Cell invasion and cell migration of NPC were inhibited due to silence of SNHG7 and were promoted due to overexpression of SNHG7. Moreover, results of further experiments revealed that the EMT-related proteins were regulated via knockdown or overexpression of SNHG7 in NPC. Furthermore, tumor metastasis of NPC was inhibited via knockdown of SNHG7 and was enhanced via overexpression of SNHG7 in nude mice. These results indicate that SNHG7 enhances NPC cell invasion and cell migration by eliciting the EMT process.

miRNA-520c-3p accelerates progression of nasopharyngeal carcinoma via targeting RAB22A.

Biological function of microRNA-20c-3p (miRNA-520c-3p) in the progression of nasopharyngeal carcinoma (NPC) and the potential mechanism were investigated. Relative level of miRNA-520c-3p in NPC tissues and adjacent normal tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Particularly, miRNA-520c-3p level in NPC with different tumor stages and tumor sizes was examined. Subsequently, miRNA-520c-3p level in nasopharyngeal epithelial cells and NPC cells was detected. The potential influence of miRNA-520c-3p on the proliferative ability and cell cycle progression of NPC cells were evaluated through cell counting kit-8 (CCK-8) and flow cytometry. The target gene of miRNA-520c-3p was verified by dual-luciferase reporter gene assay. Regulatory role of miRNA-520c-3p/RAB22A in the malignant progression of NPC was identified. miRNA-520c-3p was downregulated in NPC tissues and cell lines. Its level was lower in NPC with worse tumor grade and larger tumor size. Overexpression of miRNA-520c-3p suppressed the proliferative ability and arrested cell cycle in G0/G1 phase. RAB22A was confirmed to be the downstream target of miRNA-520c-3p. In NPC tissues and cell lines, RAB22A remained in higher abundance relative to controls. Overexpression of RAB22A reversed the inhibitory effects of overexpressed miRNA-520c-3p on proliferative ability and cell cycle progression of NPC cells. miRNA-520c-3p is downregulated in NPC, which accelerates the malignant progression of NPC by targeting RAB22A.

miRNA-21 promotes cell proliferation and invasion via VHL/PI3K/AKT in papillary thyroid carcinoma.

Papillary thyroid carcinoma (PTC) is the main kind of thyroid carcinoma, most of which are diagnosed in women. MiR-21 has been reported to be upregulated in multiple cancers to effect tumor growth. However, the role of miR-21 in PTC development remains unclear. In this present study, miR-21 and VHL expressions in PTC tissues and cells were evaluated by RT-qPCR and/or western blot. MTT assay and transwell assay were employed to assess cell proliferative and invasive abilities, respectively. Luciferase reporter assay was carried out to identify the target of miR-21and explore its roles in PTC. MiR-21 was upregulated in PTC tissues and cells. Ectopic of miR-21 expression promoted cell proliferative and invasive abilities, while knockdown miR-21 suppressed these in TPC-1 and BCPAP cells. Overexpression of miR-21 predicted poor prognosis in PTC. What is more, luciferase reporter assays showed miR-21 can directly target VHL in PTC cells. Knockdown of miR-21 expression inhibited TPC-1 and BCPAP cell invasion-mediated EMT and proliferation through the PI3K/AKT pathway. In addition, VHL reverses partial function of miR-21 on PTC cell proliferation and invasion. MiR-21 can inhibit cell proliferation and invasion by regulated VHL in PTC cells. The newly identified miR-21/VHL axis might provide a novel insight into the pathogenesis and therapy of PTC.

Identification of key genes and miRNAs markers of papillary thyroid cancer.

OBJECTIVE: In this study, crucial genes and microRNAs (miRNAs) associated with the progression, staging, and prognosis of papillary thyroid cancer (PTC) were identified. METHODS: Four PTC datasets, including our own mRNA-sequencing (mRNA-seq) dataset and three public datasets downloaded from Gene Expression Omnibus and The Cancer Genome Atlas, were used to analyze differentially expressed genes (DEGs) and miRNAs (DEMs) between PTC tumor tissues and paired normal tissues (control). Gene ontology (GO) terms and pathways associated with these DEGs were identified, and protein-protein interactions (PPIs) were analyzed. Additionally, an miRNA-mRNA regulatory network was constructed and the functions of DEMs were explored. Finally, miRNAs/mRNAs associated with tumor staging and prognosis were identified. The expression levels of several key genes and miRNAs were validated by qRT-PCR. RESULTS: Numerous DEGs and DEMs were identified between tumor and control groups in four datasets. The DEGs were significantly enriched in cell adhesion and cancer-related GO terms and pathways. In the constructed PPI network, ITGA2, FN1, ICAM1, TIMP1 and CDH2 were hub proteins. In the miRNA-mRNA negative regulatory networks, miR-204-5p regulated the largest number of target genes, such as TNFRSF12A. miR-146b, miR-204, miR-7-2, and FN1 were associated with tumor stage in PTC, and TNFRSF12A and CLDN1 were related to prognosis. CONCLUSIONS: Our results suggested the important roles of ITGA2, FN1, ICAM1, TIMP1 and CDH2 in the progression of PTC. miR-204-5p, miR-7-2, and miR-146b are potential biomarkers for PTC staging and FN1, CLDN1, and TNFRSF12A may serve as markers of prognosis in PTC.

Identification of key genes and miRNAs markers of papillary thyroid cancer

OBJECTIVE: In this study, crucial genes and microRNAs (miRNAs) associated with the progression, staging, and prognosis of papillary thyroid cancer (PTC) were identified. METHODS: Four PTC datasets, including our own mRNA-sequencing (mRNA-seq) dataset and three public datasets downloaded from Gene Expression Omnibus and The Cancer Genome Atlas, were used to analyze differentially expressed genes (DEGs) and miRNAs (DEMs) between PTC tumor tissues and paired normal tissues (control). Gene ontology (GO) terms and pathways associated with these DEGs were identified, and protein-protein interactions (PPIs) were analyzed. Additionally, an miRNA-mRNA regulatory network was constructed and the functions of DEMs were explored. Finally, miRNAs/mRNAs associated with tumor staging and prognosis were identified. The expression levels of several key genes and miRNAs were validated by qRT-PCR. RESULTS: Numerous DEGs and DEMs were identified between tumor and control groups in four datasets. The DEGs were significantly enriched in cell adhesion and cancer-related GO terms and pathways. In the constructed PPI network, ITGA2, FN1, ICAM1, TIMP1 and CDH2 were hub proteins. In the miRNA-mRNA negative regulatory networks, miR-204-5p regulated the largest number of target genes, such as TNFRSF12A. miR-146b, miR-204, miR-7-2, and FN1 were associated with tumor stage in PTC, and TNFRSF12A and CLDN1 were related to prognosis. CONCLUSIONS: Our results suggested the important roles of ITGA2, FN1, ICAM1, TIMP1 and CDH2 in the progression of PTC. miR-204-5p, miR-7-2, and miR-146b are potential biomarkers for PTC staging and FN1, CLDN1, and TNFRSF12A may serve as markers of prognosis in PTC.

Investigating the mechanisms of papillary thyroid carcinoma using transcriptome analysis.

As the predominant thyroid cancer, papillary thyroid cancer (PTC) accounts for 75­85% of thyroid cancer cases. This research aimed to investigate transcriptomic changes and key genes in PTC. Using RNA­sequencing technology, the transcriptional profiles of 5 thyroid tumor tissues and 5 adjacent normal tissues were obtained. The single nucleotide polymorphisms (SNPs) were identified by SAMtools software and then annotated by ANNOVAR software. After differentially expressed genes (DEGs) were selected by edgR software, they were further investigated by enrichment analysis, protein domain analysis, and protein­protein interaction (PPI) network analysis. Additionally, the potential gene fusion events were predicted using FusionMap software. A total of 70,172 SNPs and 2,686 DEGs in the tumor tissues, as well as 83,869 SNPs in the normal tissues were identified. In the PPI network, fibronectin 1 (FN1; degree=31) and transforming growth factor ß receptor 1 (TGFßR1; degree=22) had higher degrees. A total of 7 PPI pairs containing the non­synonymous risk SNP loci in the interaction domains were identified. Particularly, the interaction domains involved in the interactions of FN1 and 5 other proteins (such as FN1­tenascin C, TNC) had non­synonymous risk SNP loci. Furthermore, 11 and 4 gene fusion events were identified in all of the tumor tissues and normal tissues, respectively. Additionally, the NK2 homeobox 1­surfactant associated 3 (NKX2­1­SFTA3) gene fusion was identified in both tumor and normal tissues. These results indicated that TGFßR1 and the NKX2­1­SFTA3 gene fusion may be involved in PTC. Furthermore, FN1 and TNC containing the non­synonymous risk SNP loci might serve a role in PTC by interacting with each other.

[Stent implantation in the treatment of pharynx anastomotic stenosis after cervical esophageal resection: a case report].

We report the treatment of one patient with pharynx anastomotic stenosis after cervical esophagealresection by stent implantation. The patient suffered from serious pharynx anastomotic stenosis after gastric-pha-ryngeal anastomosis. After balloon-dilatation,a domestic self-expanding Z-stents was implanted in the stricture ofthe esophagus under the X-rays. After stent implantation, the patient has been leading a normal life for threeyears. Balloon dilatation and stent implantation is an effective and safe method in the treatment of patients withpharynx anastomotic stenosis.

RNA sequencing identifies crucial genes in papillary thyroid carcinoma (PTC) progression.

PURPOSE: The study aims to uncover molecular mechanisms of PTC (papillary thyroid carcinoma) progression and provide therapeutic biomarkers. METHODS: The paired tumor and control tissues were obtained from 5 PTC patients. RNA was extracted and cDNA libraries were constructed. RNA-sequencing (RNA-seq) was performed on the Illumina HiSeq2000 platform using paired-end method. After preprocessing of the RNA-seq data, gene expression value was calculated by RPKM. Then the differentially expressed genes (DEGs) were identified with edgeR. Functional enrichment and protein-protein interaction (PPI) network analyses were conducted for the DEGs. Module analysis of the PPI network was also performed. Transcription factors (TFs) of DEGs were predicted. RESULTS: A cohort of 496 up-regulated DEGs mainly correlating with the ECM degradation pathways, and 440 down-regulated DEGs predominantly enriching in transmembrane transport process were identified. Hub nodes in the PPI network were RRM2 and a set of collagens (COL1A1, COL3A1 and COL5A1), which were also remarkable in module 3 and module 5, respectively. Genes in module 3 were associated with cell cycle pathways, while in module 5 were related to ECM degradation pathways. PLAU, PSG1 and EGR2 were the crucial TFs with higher transcriptional activity in PTC than in control. CONCLUSION: Several genes including COL1A1, COL3A1, RRM2, PLAU, and EGR2 might be used as biomarkers of PTC therapy. Among them, COL1A1 and COL3A1 might exert their functions via involving in ECM degradation pathway, while RRM2 through cell cycle pathway. PLAU might be an active TF, whereas EGR2 might be a tumor suppressor.

[Application of partial laryngectomy in laryngeal cancer treatment and reconstruction with epiglottic flap and bi-pedicle muscle flap].

OBJECTIVE: To explore and evaluate the surgical method of partial laryngectomy and the laryngeal defects were reconstructed. METHOD: Two hundreds and sixty-five patients with laryngeal cancer were treated by partial laryngectomy and the laryngeal defects were reconstructed by epiglottic flap, bi-pedicle sternohyoid muscle fascia flap and epiglottic flap plus bi-pedicle sternohyoid muscle fascia flap from January 1990 to December 2000. There were 256 males and 9 females,ranging in age from 48 to 75 years old. Neck dissection were performed in 79 patients. RESULT: The total cases began to eat between 10 to 15 days after operation. Two hundreds and eighteen cases were decannulated with a decannulation rate of 82.26%. Wound infection occurred in 12 cases and 2 pharyngocutaneous fistula was found. The three and five years survival rates were 74.72% and 70. 88% respectively. CONCLUSION: After partial laryngectomy in laryngeal cancer epiglottic flap and bi-pedicle muscle fascia flap were performed reconstruction of laryngeal function in a single-stage. These procedures which were relative simplicity, expediency, small injury and high rate of survival present ideal effects and are worthy to be recommended.