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Objective: Atherosclerosis (AS) is the leading cause of death for humans worldwide, and some circular RNAs (circRNAs) have been demonstrated to play important roles in its progression. In this study, we mainly investigated the functions and molecular mechanisms of circRNA-PTPRA (circPTPRA) in AS., Patients and Methods: The expressions of circPTPRA and miR-636 were detected in serum samples of AS patients (n=30) and healthy controls (n=30) by RT-PCR. Then levels of circPTPRA were detected after ox-LDL treatment into vascular smooth muscle cell (VSMCs), macrophage and endothelial cells. LV-sh circPTPRAs were constructed and infected into VSMCs. CCK-8 assay was performed to measure cell proliferation abilities, flow cytometry (FACS) was performed to measure cell-cycle distribution and TUNEL staining was performed to detect cell apoptosis. Western blot (WB) was performed to detect protein levels of SP1, Cyclin D1, Cyclin E, Bax, Bad, Cleaved Caspase3. Luciferase reporter assay was performed to verify the potential binding sites of circPTPRA and miR-636, miR-636 and SP1., Results: RT-PCR showed that circPTPRA was upregulated in serum samples of AS patients, which was increased by ox-LDL in VSMCs. CircPTPRA inhibition repressed cell proliferation, improved cell-cycle distribution in G0/G1 phase and promoted cell apoptosis. MiR-636, a potential target for circPTPRA, was reduced in serum samples of AS patients and Luciferase reporter assay confirmed that circPTPRA could directly sponge with miR-636 in VSMCs. Furthermore, miR-636 inhibition promoted proliferation and repressed apoptosis of VSMCs, while miR-636 overexpression reversed these results. SP1, a transcription factor that played some roles in the progression of AS, was predicted to be a target of miR-636. MiR-636 inhibition increased SP1 while miR-636 overexpression repressed SP1 expression, Luciferase reporter assay proved that miR-636 could target at SP1 in VSMCs. Moreover, the repressed cell proliferation and promoted cell apoptosis abilities in LV-sh SP1 were reversed following with miR-636 inhibitor transfection. In addition, the repressed cell proliferation and promoted cell apoptosis abilities in VSMCs with LV-sh circPTPRAs were reversed following with miR-636 inhibitor transfection, which suggested that circPTPRA regulated cell proliferation and apoptosis through miR-636/SP1 axis in AS., Conclusions: According to the results, we found that circPTPRA was upregulated in serum samples of AS patients, which promoted cell proliferation and inhibited cell apoptosis through repressing miR-636 and upregulating SP1 signaling axis. Our results uncovered a potential role of circPTPRA, which might be a marker and therapeutic target for AS patients.

Objective: Micro-ribonucleic acids (miRNAs) are involved in the occurrence of various cancers, and the hypoxia-inducible factor 1-α (HIF-1α) is the main regulator of cell proliferation induced by hypoxia. The relationships of miR-199a and HIF-1α expressions with non-small cell lung cancer (NSCLC) remain unclear, so they were explored in this work., Materials and Methods: On the basis of establishing the rat model of NSCLC, the messenger RNA (mRNA) expressions of miR-199a, HIF-1α and the vascular endothelial growth factor (VEGF) were analyzed in NSCLC rats, and the correlations of miR-199a with the mRNAs of HIF-1α and VEGF and cancer staging were investigated. To further study the role of miR-199a in NSCLC cell proliferation via the HIF-1α/VEGF signaling pathway, NSCLC cells were treated with the signaling pathway inhibitor and transfected with miR-199a mimics, respectively. Also,  the roles of the inhibitor PX-478 and miR-199a mimics in the expressions of miR-199a, HIF-1α, and VEGF proteins, as well as their influences on cell proliferation ability were detected., Results: In NSCLC rats, the expression of miR-199a was substantially decreased (p<0.01), but the expressions of HIF-1α and VEGF were notably raised (p<0.01). MiR-199a was negatively correlated with the expression of VEGF. As cancer developed, the expression of miR-199a was gradually lowered, but the expressions of HIF-1α and VEGF were gradually increased. Both HIF-1α/VEGF signaling pathway inhibitor PX-478 and miR-199a mimics significantly reduced the expressions of HIF-1α and VEGF proteins (p<0.01) and suppressed the cell proliferation activity., Conclusions: MiR-199a prevents the proliferation of NSCLC cells via the targeted down-regulation of the HIF-1α/VEGF signaling pathway.

Objective: This study aims to investigate whether SNHG16 (small nucleolar RNA host gene 16) can promote the progression of osteoarthritis (OA) by regulating the microRNA-93-5p/Cyclin D1 (CCND1) axis, thereby finding new therapeutic targets for the treatment of OA., Patients and Methods: A total of 23 OA patients and 23 patients undergoing lower extremity amputation were enrolled in this study. We collected their cartilage tissues from knee joint for isolating chondrocytes. The relative levels of SNHG16, CCND1 and microRNA-93-5p in cartilage tissues of OA patients and controls were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The regulatory effect of SNHG16 on proliferative potential of chondrocytes was evaluated by Cell Counting Kit-8 (CCK-8) and colony formation assay, respectively. Cell cycle progression was examined using flow cytometry. Dual-Luciferase reporter gene assay was conducted to verify the binding between SNHG16 with microRNA-93-5p and microRNA-93-5p with CCND1. Rescue experiments were performed to elucidate whether SNHG16 regulated CCND1 expression by targeting microRNA-93-5p., Results: The expressions of SNHG16 and CCND1 upregulated, while microRNA-93-5p downregulated in cartilage tissues of OA patients relative to controls. Correlation regression analyses showed a negative expression correlation between SNHG16 and microRNA-93-5p, as well as CCND1 and microRNA-93-5p in OA patients. On the contrary, SNHG16 expression was positively correlated to CCND1 expression in OA. The knockdown of SNHG16 suppressed viability, cloning ability and cell cycle progression, but induced apoptosis in chondrocytes. Dual-Luciferase reporter gene assay showed that SNHG16 could bind to microRNA-93-5p. SNHG16 knockdown markedly upregulated the expression of microRNA-93-5p. Moreover, the knockdown of microRNA-93-5p reversed the inhibited viability due to SNHG16 knockdown. Transfection of microRNA-93-5p mimics markedly inhibited CCND1 expression. Importantly, CCND1 overexpression reversed the inhibitory effect of SNHG16 knockdown on chondrocyte viability., Conclusions: SNHG16 promotes the development of OA by regulating microRNA-93-5p/CCND1 axis.

Objective: The aim of this study was to explore whether maternally expressed gene 3 (MEG3) could facilitate the proliferation and migration of oral squamous cell carcinoma (OSCC) cells by selectively binding to miR-21, thereby participating in the progression of OSCC., Patients and Methods: The expression levels of MEG3 and miR-21 in OSCC tissues and normal control tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The effects of MEG3 and miR-21 on cell proliferation and migration were examined by cell counting kit-8 (CCK-8), transwell, and scratch assay, respectively. Meanwhile, cell cycle was detected using flow cytometry. The binding relationship between miR-21 and MEG3 was confirmed by dual luciferase assay. In addition, MEG3 and miR-21 were simultaneously knock-down to figure out whether MEG3 could regulate the proliferation and migration of OSCC cells through targeted binding to miR-21., Results: QRT-PCR results indicated that MEG3 expression in OSCC tissues was remarkably lower than that of normal control tissues. However, the expression of miR-21 was significantly higher in OSCC tissues. Meanwhile, it was found that inhibiting MEG3 expression in OSCC cell lines could significantly promote cell proliferation and migration, while the simultaneous inhibition of miR-21 showed the opposite effect. Dual Luciferase assay results revealed that MEG3 could selectively bind to miR-21. In addition, we demonstrated that the knockdown of MEG3 in Tca-8113 and CAL-27 cells partially reversed the inhibitory effect of downregulated-miR-21 on cell proliferation and migration. These results further suggested that MEG3 might regulate OSCC cell proliferation via selectively binding to miR-21., Conclusions: Low expression of MEG3 can promote the proliferation and migration of OSCC cells through targeted binding to miR-21.

Objective: To investigate the impact of miR-124Aa on the proliferation, invasion and cytokine excretion of rheumatoid arthritis synovial fibroblasts (RASFs) in patients with rheumatoid arthritis (RA)., Patients and Methods: RASFs were separated for in-vitro culture, and transfected using lipidosome that connected with chemically synthesized miR-124a mimic or miR-124a inhibitor. Then, MTT, transwell chamber, and flow-cytometry were used to detect the impact on the proliferation, invasion, and apoptosis of RASFs; RT-PCR and Western-blotting were employed to measure the effect of miR-124a on the expressions of matrixmetalloproteinase3/13 (MMP3/13) and interleukin1β (IL-1β) of RASFs., Results: miR-124a significantly suppresses the proliferation of RASFs, while inhibits the invasion of RASFs. The flow cytometry indicated that miR-124a showed no significant effect on the apoptosis of RASFs. Finally, miR-124a downregulates the expressions of MMP3/13 and IL-1β., Conclusions: MiR-124a is of great significance for the onset of RA by inhibiting the proliferation and invasion of RASFs possibly through downregulating the expression of MMP3/13 and IL-1β.

Objective: Previous study has reported that miR-330-3p was highly expressed in breast cancer (BC) patients. However, the effect of miR-330-3p in BC progression remains largely unclear. The purpose of this study was to investigate the clinical significance of miR-330-3p expression in BC., Patients and Methods: The expression of miR-330-3p was detected by quantitative Real-time PCR in BC tissues and matched normal breast tissues. The association of miR-330-3p expression with clinicopathological factors of BC patients was also analyzed by x2-test. Prognosis value of patients with BC was assessed by Kaplan-Meier method and Cox proportional hazards model, respectively., Results: Quantitative real-time PCR analysis showed that the expression level of miR-330-3p was significantly higher in BC specimens than that in corresponding noncancerous tissues (p < 0.01). The levels of miR-330-3p were positively correlated with the status of TNM stage (p = 0.011) and lymph node metastasis (p = 0.006). Kaplan-Meier analysis revealed that 5-year overall survival of BC patients with high miR-330-3p expression was shorter compared to those patients with low miR-330-3p expression (p < 0.0001). Moreover, univariate and multivariate regression analysis demonstrated that miR-330-3p was an independent prognostic factor in BC., Conclusions: Our data suggest that miR-330-3p upregulation maybe concurrently associated with prognosis in patients with BC, suggesting that miR-330-3p may be a potential prognostic biomarker and therapeutic target for patients with BC.

Objective: Myocardial cell apoptosis represents important pathologic basis of ischemia-reperfusion injury (I/R). MiR-23a is related to myocardial hypertrophy and cardiac remodeling by regulating myocardial cell growth and apoptosis. This study intended to observe the regulating effect of miR-23a in myocardial cell and related target, and investigate its clinical significance to I/R injury., Materials and Methods: The rats were divided into sham group and myocardial I/R group. Myocardial cell cycle and miR-23a expression were tested. H2O2 was applied to treat H9c2 rat myocardial cell to simulate oxidative stress during I/R. The cells were divided into blank group, NC group, miR-23a mimic group, H2O2 group, and miR-23a + H2O2 group. ROS content and cell apoptosis were detected by flow cytometry. MiR-23a, FoxO3a, and BIM gene expression were determined by qRT-PCR. FoxO3a and BIM protein levels were measured by Western blot., Results: Compared with sham group, myocardial apoptosis increased, while miR-23a expression was significantly downregulated in I/R group. H2O2 treatment markedly increased ROS levels in H9c2 cells and elevated apoptosis. The overexpression of mMiR-23a effectively reduced cell apoptosis induced by H2O2 treatment. H2O2 treatment significantly decreased miR-23a expression, while markedly elevated the levels of FoxO3a and BIM. The overexpression of miR-23a apparently impeded the induction of FoxO3a and BIM by H2O2., Conclusions: The downregulation of miR-23a plays a negative role in oxidative stress and cell apoptosis induced by I/R. The overexpression of miR-23a is of significance to alleviate cell apoptosis through inhibiting FoxO3a and downstream target BIM expression.

Objective: Colorectal cancer (CRC) is a common human malignancy and is the second leading cause of cancer deaths worldwide with a dismal prognosis. Previous investigations have shown that miR-340 can modulate the metabolism of CRC cells. The aim of this report is to study the role of miR-340 in the development and progression of CRC., Patients and Methods: The level of miR-340 in CRC cells was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting. CRC cell lines were used as model cell lines and the anti-tumor effect of miR-340 in vitro was examined. The luciferase reporter assay was performed. The level of miR-340 was restored in CRC cells by the usage of the miR-340 mimic. Re-expression of RLIP76 in CRC cells was then constructed. Moreover, the target gene of miR-340 was identified through the experiment of in vivo xenograft model., Results: The aberrant downregulation of miR-340 is correlated with advanced stage of CRC. Furthermore, the ectopic overexpression of miR-340 in CRC cell lines resulted in growth inhibition, apoptosis and enhanced chemosensitivity in vitro and in vivo, which was mediated by directly targeting RLIP76., Conclusions: miR-340 acts as a tumor suppressor in CRC and is involved in the chemoresistance of CRC.

Trichinella spiralis is an important zoonotic nematode causing trichinellosis which is associated with human diseases such as malaise, anorexia, nausea, vomiting, abdominal pain, fever, diarrhea, and constipation. microRNAs (miRNAs) are endogenous small non-coding RNAs that play important roles in the regulation of gene expression. The objective of the present study was to examine the miRNA expression profile of the larvae of T. spiralis by Solexa deep sequencing combined with stem-loop real-time polymerase chain reaction (PCR) analysis. T. spiralis larvae were collected from the skeletal muscle of naturally infected pigs in Henan province, China, by artificial digestion using pepsin. The specific identity of the T. spiralis larvae was confirmed by PCR amplification and subsequent sequence analysis of the internal transcribed spacer of ribosomal DNA. A total of 17,851,693 reads with 2,773,254 unique reads were obtained. Eleven conserved miRNAs from 115 unique xsmall RNAs (sRNAs) and 12 conserved miRNAs from 130 unique sRNAs were found by BLAST analysis against the known miRNAs of Caenorhabditis elegans ( ftp://ftp.ncbi.nih.gov/genomes/Caenorhabditis&#95;elegans ) and Brugia malayi dataset ( http://www.ncbi.nlm.nih.gov/genomeprj?Db=genomeprj&cmd=ShowDetailView&TermToSearch=9549 ) in miRBase, respectively. One novel miRNA with 12 precursors were identified and certified using the reference genome of B. malayi, while no novel miRNA was found when using the reference genome of C. elegans. Nucleotide bias analysis showed that the uracil was the prominent nucleotide, particularly at the 1st, 6th, 18th, and 23th positions, which were almost at the beginning, middle, and the end of the conserved miRNAs. The identification and characterization of T. spiralis miRNAs provides a new resource to study regulation of genes and their networks in T. spiralis.