Construction of prognostic risk model of bladder cancer based on cuproptosis-related long non-coding RNAs / 浙江大学学报·医学版

OBJECTIVES@#To construct a prognosis risk model based on long noncoding RNAs (lncRNAs) related to cuproptosis and to evaluate its application in assessing prognosis risk of bladder cancer patients.@*METHODS@#RNA sequence data and clinical data of bladder cancer patients were downloaded from the Cancer Genome Atlas database. The correlation between lncRNAs related to cuproptosis and bladder cancer prognosis was analyzed with Pearson correlation analysis, univariate Cox regression, Lasso regression, and multivariate Cox regression. Then a cuproptosis-related lncRNA prognostic risk scoring equation was constructed. Patients were divided into high-risk and low-risk groups based on the median risk score, and the immune cell abundance between the two groups were compared. The accuracy of the risk scoring equation was evaluated using Kaplan-Meier survival curves, and the application of the risk scoring equation in predicting 1, 3 and 5-year survival rates was evaluated using receiver operating characteristic (ROC) curves. Univariate and multivariate Cox regression were used to screen for prognostic factors related to bladder cancer patients, and a prognostic risk assessment nomogram was constructed, the accuracy of which was evaluated with calibration curves.@*RESULTS@#A prognostic risk scoring equation for bladder cancer patients was constructed based on nine cuproptosis-related lncRNAs. Immune infiltration analysis showed that the abundances of M0 macrophages, M1 macrophages, M2 macrophages, resting mast cells and neutrophils in the high-risk group were significantly higher than those in the low-risk group, while the abundances of CD8+ T cells, helper T cells, regulatory T cells and plasma cells in the low-risk group were significantly higher than those in the high-risk group (all P<0.05). Kaplan-Meier survival curve analysis showed that the total survival and progression-free survival of the low-risk group were longer than those of the high-risk group (both P<0.01). Univariate and multivariate Cox analysis showed that the risk score, age and tumor stage were independent factors for patient prognosis. The ROC curve analysis showed that the area under the curve (AUC) of the risk score in predicting 1, 3 and 5-year survival was 0.716, 0.697 and 0.717, respectively. When combined with age and tumor stage, the AUC for predicting 1-year prognosis increased to 0.725. The prognostic risk assessment nomogram for bladder cancer patients constructed based on patient age, tumor stage, and risk score had a prediction value that was consistent with the actual value.@*CONCLUSIONS@#A bladder cancer patient prognosis risk assessment model based on cuproptosis-related lncRNA has been successfully constructed in this study. The model can predict the prognosis of bladder cancer patients and their immune infiltration status, which may also provide a reference for tumor immunotherapy.

Huangqi Decoction, a compound Chinese herbal medicine, inhibits the proliferation and activation of hepatic stellate cells by regulating the long noncoding RNA-C18orf26-1/microRNA-663a/transforming growth factor-β axis / 中西医结合学报

OBJECTIVE@#Huangqi Decoction (HQD), a classical traditional Chinese medicine formula, has been used as a valid treatment for alleviating liver fibrosis; however, the underlying molecular mechanism is still unknown. Although our previous studies showed that microRNA-663a (miR-663a) suppresses the proliferation and activation of hepatic stellate cells (HSCs) and the transforming growth factor-β/small mothers against decapentaplegic (TGF-β/Smad) pathway, whether long noncoding RNAs (lncRNAs) are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported. The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.@*METHODS@#The expression levels of lnc-C18orf26-1, miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction. HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs. The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs. Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs. RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs. Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA. The expression levels of collagen α-2(I) chain (COL1A2), α-smooth muscle actin (α-SMA) and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.@*RESULTS@#Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a. Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation, downregulated TGF-β1-stimulated α-SMA and COL1A2 expression, and inhibited the TGF-β1/Smad signaling pathway. HQD suppressed the proliferation and activation of HSCs. HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs. Further studies showed that HQD inhibited the expression of COL1A2, α-SMA, TGF-β1, TGF-β type I receptor (TGF-βRI) and phosphorylated Smad2 (p-Smad2) in HSCs, and these effects were reversed by miR-663a inhibitor treatment.@*CONCLUSION@#Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis. HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.

Regulation of Glial Function by Noncoding RNA in Central Nervous System Disease / 神经科学通报·英文版

Non-coding RNAs (ncRNAs) are a class of functional RNAs that play critical roles in different diseases. NcRNAs include microRNAs, long ncRNAs, and circular RNAs. They are highly expressed in the brain and are involved in the regulation of physiological and pathophysiological processes of central nervous system (CNS) diseases. Mounting evidence indicates that ncRNAs play key roles in CNS diseases. Further elucidating the mechanisms of ncRNA underlying the process of regulating glial function that may lead to the identification of novel therapeutic targets for CNS diseases.

Identification of LHFPL3-AS2 as a prognostic biomarker in lung adenocarcinoma

Background: Deregulation of long noncoding RNAs (lncRNAs) was considered one of the main characteristics of several human cancers. However, detailed genome-wide expression and functional significance studies of lnc RNAs in lung adenocarcinoma are still limited. This study aims to discover a new lncRNA that may play an important role in regulating the pathogenesis of lung adenocarcinoma (ADC). Methods: We conducted a comprehensive analysis of three Gene Expression Omnibus (GEO) microarray datasets and TCGA datasets. Differentially expressed lncRNAs between ADC and normal tissues were screened and verified using Gene Expression Profiling Interactive Analysis (GEPIA). Moreover, Kaplan-Meier plotter was used to construct the gene prognosis profile. The downstream targets of miRNA and related functional pathways were predicted and validated. Results: With microarray gene expression analysis, we found that only lncRNAs-PCAT6 was commonly upregulated among four datasets, and four lncRNAs (LINC00968, PGM5-AS1, LHFPL3-AS2 and SFTA1P) were significantly downregulated in the ADC samples as compared to the normal tissues. Meanwhile, for LHFPL3-AS2, high-risk patients showed better overall survival (HR=0.6 or 0.62; P < .0001 or P = 0.0014), overall survival from TCGA datasets (HR=0.72; P = 0.015) and recurrence-free survival (HR=0.72; P = 0.015). Then, LHFPL3-AS2 was predicted to bind to two miRNAs, miR-127-5p and miR-424-5p. Finally, validation and functional enrichment analysis of the downstream key mRNAs showed significant enrichment in some cancer-related pathways, such as cell adhesion in cancer and small cell lung cancer. Conclusions: Taken together, our study indicated that LHFPL3-AS2 was associated with tumorigenesis, and it could be used as a useful biomarker in the diagnosis, prognosis and treatment of ADC.

Diagnostic and prognostic significance of long noncoding RNA LINC00173 in patients with melanoma

SUMMARY OBJECTIVE: A growing volume of literature has suggested long noncoding RNAs (lncRNAs) as important players in tumor progression. In this study, we aimed to investigate the expression and prognostic value of lncRNA LINC00173 (LINC00173) in melanoma. METHODS: LINC00173 expression was measured in 163 paired cancerous and noncancerous specimen samples by real-time polymerase chain reaction. The correlations between LINC00173 expression with clinicopathological characteristics and prognosis were analyzed by chi-square test, log-rank test, and multivariate survival analysis. Receiver-operating characteristic curves were used for the assessment of the diagnostic value of LINC00173 for melanoma patients. RESULTS: The expression level of LINC00173 in melanoma specimens was distinctly higher than that in adjacent non-neoplasm specimens (p<0.01). Besides, LINC00173 was expressed more frequently in patients with advanced melanoma than in patients with early melanoma. Multivariate assays confirmed that LINC00173 expression level was an independent prognostic predictor of melanoma patients (p<0.05). CONCLUSION: Our data indicated that LINC00173 expression could serve as an unfavorable prognostic biomarker for melanoma patients.

Study on long noncoding RNA LBX2-AS1 regulates glioma cell proliferation, migration and apoptosis through epidermal growth factor receptor signaling pathway / 中国医师进修杂志

Objective:To investigate the mechanism of long noncoding RNA (lncRNA) LBX2-AS1 regulating glioma cell proliferation, migration and apoptosis through epidermal growth factor receptor (EGFR) signaling pathway.Methods:From April 2018 to August 2021, glioma U251 cells (U251 cells for short) were divided into control group and observation group, with 4 strains in each group. The control group was routinely cultured, and the observation group was transfected with specific small interfering RNA (siRNA) targeting LBX2-AS1. The proliferation ability of U251 cells was detected by methyl thiazol tetrazolium method, the metastasis rate of U251 cells was detected by scratch test, the apoptosis rate of U251 cells was detected by flow cytometry, and the expression of total protein and vascular endothelial growth factor (VEGF), phosphorylated inositol 3 kinase (p-PI3K), phosphorylated protein kinase B (p-Akt), phosphorylated Ras (p-Ras) and phosphorylated Raf (p-Raf) protein were detected by Western blot.Results:The proliferation ability and metastasis rate of U251 cells in observation group were significantly lower than those in control group: (27.15 ± 1.38)% vs. (63.54 ± 2.47)% and (37.09 ± 3.74)% vs. (82.17 ± 9.24)%, the apoptosis rate of U251 cells was significantly higher than that in control group: (69.17 ± 5.83)% vs. (17.58 ± 1.22)%, and there were statistical differences ( P<0.01). The expression of total protein and VEGF, p-PI3K, p-Akt, p-Ras, p-Raf protein of U251 cells in observation group were significantly lower than those in control group (1.52 ± 0.23 vs. 2.39 ± 0.31, 0.73 ± 0.08 vs. 1.68 ± 0.45, 0.57 ± 0.11 vs. 1.89 ± 0.31, 0.68 ± 0.06 vs. 1.74 ± 0.51, 0.84 ± 0.12 vs. 1.99 ± 0.63 and 0.71 ± 0.08 vs. 1.52 ± 0.37), and there were statistical differences ( P<0.01). Conclusions:The lncRNA LBX2-AS1 is highly expressed in glioma cells. Silencing the expression of lncRNA LBX2-AS1 inhibits the proliferation and metastasis of glioma cells through EGFR pathway.

LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Long noncoding RNAs (lncRNAs) are expressed in different species and different tissues, and perform different functions, but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone (FSH). In general, we have revealed lncRNA‍‒‍microRNA (miRNA)‍‒‍‍messenger RNA (mRNA) interactions that may play important roles in rat primary pituitary cells. In this study, a new lncRNA was identified for the first time. First, we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization, which indicated that this lncRNA was distributed mainly in the cytoplasm. Next, we used RT-qPCR and enzyme-linked immunosorbent assay (ELISA) to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion. In addition, mothers against decapentaplegic homolog 2 (Smad2) was highly expressed in our sequencing results. We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2. We used RNA immunoprecipitation-qPCR (RIP-qPCR) and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2. Fluorescence in situ hybridization (FISH) showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm. Finally, we determined the relationship among lncRNA-m18as1, miR-18a-5p, and the Smad2/3 pathway. Overall, we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.

Neuroprotective effect of long noncoding RNA-nuclear-enriched abundant transcript 1 on traumatic brain injury in mice and its possible mechanism / 中华创伤杂志

Objective:To explore the impact of the expression of long noncoding RNA-nuclear-enriched abundant transcript 1 (NEAT1) on neurological function and neuronal apoptosis after traumatic brain injury (TBI) in mice and the possible mechanism.Methods:According to the random number table, 90 C57BL/6J mice were divided into sham group, blank control group, empty virus group 1, empty virus group 2, NEAT1 over-expression group and NEAT1 knockdown group, with 15 mice per group. The traumatic brain injury (TBI) was simulated by controlled cortical injury (CCI) model, and NEAT1 was regulated by intracerebroventricular injection with recombinant adenovirus. The neurological severity score (NSS) and Morris water maze test were used to evaluate the neurological function in blank control group, NEAT1 over-expression group and NEAT1 knockdown group within 1 week and 14-21 days after injury. The Western blot was used to observe the expressions of P53-induced protein with a death domain 1 (Pidd1), caspase-2, caspase-9 and caspase-3 in blank control group at 6 hour and 1, 3, 7 days after injury. The TUNEL method and immunofluorescence were used to observe the neurological apoptosis and expression of Pidd1 in blank control group, NEAT1 over-expression group and NEAT1 knockdown group at 3 days after injury. The Western blot analysis was used to detect protein expressions of Pidd1, caspase-2, cytochrome c (Cyt c) and caspase-3 in sham group, blank control group, empty virus groups, NEAT1 over-expression group and NEAT1 knockdown group at 3 days after injury.Results:The NSS was significantly reduced in NEAT1 over-expression group [(3.5±0.7)points], and was significantly increased in NEAT1 knockdown group [(5.0±1.5)points]at day 1 after injury, when compared with blank control group [(4.9±1.0)points]( P<0.01). The Morris water maze test showed that the time to find platform was decreased in NEAT1 over-expression group[(10.9±2.8)seconds], and was prolonged in NEAT1 knockdown group [(30.7±6.2)seconds] at day 19 after injury ( P<0.05 or 0.01), when compared with blank control group [(20.1±5.6)seconds]. The Western blot analysis showed that the expressions of Pidd1, caspase-2, caspase-9 and caspase-3 had significant increase at day 3 after injury ( P<0.01). The TUNEL test showed that the apoptosis rate of neurons was significantly decreased in NEAT1 over-expression group [(18.0±2.7)%], and the apoptosis rate was significantly increased in NEAT1 knockdown group [(63.0±8.6)%] at day 3 after injury ( P<0.01). Immunofluorescence showed that the expression of Pidd1 protein in cytoplasm of the neurons was decreased in NEAT1 over-expression group [(22.7±2.2)%]( P<0.01), and was increased in NEAT1 knockdown group [(72.7±7.0)%]( P<0.01) at day 3 after injury, when compared with blank control group. The Western blot analysis showed that the expressions of Pidd1, capsase-2, Cyt c and caspase-3 were significantly reduced in NEAT1 over-expression group (0.5±0.0, 0.3±0.0, 0.5±0.0, 0.4±0.0) at day 3 after injury, when compared with blank control group. However, the results were opposite in NEAT1 knockdown group. Conclusion:After TBI, the NEAT1 can reduce the activation of caspase-3 through the Pidd1-caspase-2-Cyt c pathway after TBI, regulate neuronal apoptosis, and ultimately play a protective role in neurological function.

Long noncoding RNA small nucleolar RNA host gene 1 promotes gemcitabine resistance by targeting microRNA330 in pancreatic cancer cells / 中华胰腺病杂志

Objective:To investigate the role of long noncoding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1) in the resistance of pancreatic cancer PANC1 cells to gemcitabine (GEM), and clarify the relationship between SNHG1 and miR-330.Methods:The clinical data of 179 pancreatic tissue samples and 171 adjacent normal pancreas tissues were collected from the Cancer Genome Atlas (TCGA), and bioinformatics analysis was performed to analyze the expression of SNHG1 and miR-330 in pancreatic cancer tissue and adjacent normal tissue. PANC1 cell line with the resistance to GEM was established by the intermittent gradient doubling method in vitro. The GEM-resistant cells were divided into three groups: si-NC-GEM (negative control) group, si-SNHG1-GEM (transfected with siRNA targeting SNHG1) group, and GEM-resistant group. At the same time, to validate the role of miR-330 in targeting SNHG1 expression, the GEM-resistant cells were further divided into three groups: NC-inh+ si-NC group (co-transfected with negative control miR-330 inhibitor and si-NC), NC-inh+ si-SNHG1 group (co-transfected with negative control miR-330 inhibitor and si-SNHG1), and miR-330-inh + si-SNHG1 group (co-transfected with miR-330 inhibitor and si-SNHG1). qRT-PCR was used to detect the expression of SNHG1 and miR-330 in GEM-resistant cells. CCK-8 and immunofluorescence (Ki67) were used to test GEM-resistant cell growth and proliferation. Transwell and wound healing were used to test the migration and invasion in GEM-resistant cells. Bioinformatics analysis and luciferase reporter assay were used to verify the regulatory relationship between SNHG1 and miR-330. Results:Compared to the adjacent normal tissues of pancreatic cancer, the expression of lncRNA SNHG1 in pancreatic cancer tissues was significantly increased (6.543±0.72 vs 5.31±0.96), and the expression of miR-330 was greatly decreased (2.54±1.85 vs 3.01±1.23); and all the differences were statistically significant ( P<0.05). The expression of lncRNA SNHG1 in si-NC-GEM group, si-SNHG1-GEM group, and GEM-resistant group was 2.43±0.10, 0.26±0.08 and 3.25±0.310, which in si-SNHG1-GEM group was lower than those in si-NC-GEM group or GEM-resistant group. While the expression of miR-330 in si-NC-GEM group, si-SNHG1-GEM group, and GEM-resistant group was 0.47±0.13, 0.84±0.12 and 0.38±0.21, which in si-SNHG1-GEM group was higher than those in si-NC-GEM group or GEM-resistant group. All the differences were statistically significant ( P<0.05). Compared to the si-NC-GEM group or GEM-resistant group, A450, Ki67, number of migrated cells and the distance of invasion in si-SNHG1-GEM group were all decreased, and the differences were statistically significant ( P<0.05). Otherwise, luciferase activity of miR-330-WT in si SNHG1-GEM group was significantly higher than that in NC siRNA group (3.21±0.22 vs 1.03±0.18). The luciferase activity of SNHG1-WT in miR-330 inhibitor group was significantly lower than that in NC inhibitor group (0.97±0.21 vs 2.32±0.17). In miR-330-inh+ si-SNHG1 GEM-resistant group, the cell A450, Ki67, migration cell and distant of invasion was higher than those in NC-inh+ si-SNHG1 group, and the difference was statistically significant (all P value<0.05). Conclusions:lncRNA SNHG1 targeting miR-330 could promote GEM resistance in pancreatic cancer PANC1 cells.

Expression of lncRNA Tsix in a mouse model of N-methyl-D-aspartic acid-induced retinal excitotoxicity and its significance / 中华实验眼科杂志

Objective:To investigate the damage effect of different concentrations of N-methyl-D-aspartic acid (NMDA) to retinal ganglion cells (RGCs) in mice and explore the expression of long noncoding RNA (lncRNA) Tsix in the retina of mice with excitotoxicity as well as the protective effect of lncRNA Tsix on retina and RGCs.Methods:A total of 105 C57B6/J mice at 7-8 weeks of age were selected and randomly divided into the normal control group, 2 mmol/L NMDA group, 10 mmol/L NMDA group, 20 mmol/L NMDA group and 40 mmol/L NMDA group using a random number table method, with 21 mice in each group.In the normal control group, the mice were intravitreally injected with 1 μl of sodium chloride solution in the right eye, and mice were given intravitreal injection of 1 μl of different doses of NMDA according to grouping.At one week after the injection, the thickness of each retinal layer, the number of ganglion cell layer (GCL) cells and the number of RGCs were analysed and compared among different groups through optical coherence tomography (OCT), hematoxylin-eosin staining, retinal whole mount staining and immunofluorescence staining.RNAscope in situ hybridization was used to verify the expression of lncRNA Tsix in the GCL of different groups.The quantitative real-time PCR was used to detect the transcript levels of Tsix in different groups.This study was approved by an Ethics Committee of Tianjin Medical University (No.SYXK2018-0004), and the use of experimental animals was in accordance with the regulations of Tianjin Medical University and ARVO statement. Results:The OCT results showed that the total retinal thickness of mice in the 2, 10, 20 and 40 mmol/L NMDA groups were (255.00±6.63), (252.40±6.41), (248.67±6.20) and (229.11±10.37)μm, respectively, which were thinner than (269.60±20.01)μm in the normal control group, and the differences were statistically significant (all at P<0.05). Hematoxylin-eosin staining showed that the cells in the GCL of the normal control group were uniform and compact, and arranged in a single layer with large and round nuclei.In the NMDA groups, the cells were uneven in volume with vacuoles and nuclear pyknosis.The cell density in the GCL was decreased significantly with the increasing NMDA doses in NMDA groups in comparison with the normal control group, and the differences were statistically significant (all at P<0.05). In the 20 mmol/L NMDA group, the cell density in the GCL was reduced to half of the normal control group.The results of retinal whole mount staining showed that the density of β3-tubulin-positive RGCs was decreased significantly as the dose of NMDA increased in NMDA groups, and the differences were statistically significant compared with the normal control group (all at P<0.05). The number of RGCs in the 10 mmol/L NMDA group was reduced to half of that in the normal control group.RNAscope results showed that lncRNA Tsix was mainly expressed in the cytoplasm of the GCL cells.The proportion of lncRNA Tsix-positive cells was significantly reduced with the increase of the NMDA dose ( F=13.670, P<0.01). The quantitative real-time PCR results verified that the trend of Tsix expression was consistent with the RNAscope result. Conclusions:NMDA exerts a dose-dependent damage to the layer thickness of mouse retina and RGCs.The expression of lncRNA Tsix in mouse retina is mainly enriched in the cytoplasm of the cells in the GCL, and the transcript level of Tsix is reduced with the increase of NMDA concentration and have a protective effect on RGCs.

Prognostic value and potential mechanism for the role of SNHG4 in gastric cancer / 中华内分泌外科杂志

Objective:To investigate the expression, prognostic value and potential mechanism for the role of SNHG4 in gastric cancer.Methods:The expression of SNHG4 in gastric cancer was analyzed by UALCAN database. The relationship between SNHG4 and prognosis of gastric cancer was analyzed by Kaplan-Meier Plotter. SNHG4-miRNA-mRNA regulatory network was constructed by StarBase, Targetscan, microT-CDS and Cytoscape. The target genes were analysis GO and KEGG pathway enrichment by DAVID database.Results:The expression of SNHG4 in gastric cancer was significantly higher than that in normal tissues ( P=8.882E-16) . The overall survival time of patients with high SNHG4 expression was lower than that of patients with low expression ( P=8.900E-05) . Through the construction of RNA regulatory network, we found that hsa-let-7a-5p ( P=1.02E-03) , hsa-miR-152-3p ( P=4.51E-06) , hsa-miR-204-5p ( P=6.68E-04) and hsa-miR-363-3p ( P=8.06E-03) could be used as the binding sites of SNHG4 in gastric cancer, and these four miRNAs further regulated 250 downstream target genes. Through GO and KEGG enrichment analysis of the target genes, we found that these target genes played roles in the biological process of protein phosphorylation regulation, transcription negative regulation, RNA polymerase II promoter transcription, and participated in the occurrence and development of gastric cancer by blocking or activating Wnt and other signal pathways. Conclusions:SNHG4 can be used as a potential tumor marker for gastric cancer to judge the prognosis of gastric cancer. By constructing a SNHG4-miRNA-mRNA regulatory network, the pathogenesis of gastric cancer can be studied at the molecular level. This provides a clear direction for experimental and clinical research.

The long-chain noncoding RNA FAM201A influences the biological behavior and radiation sensitivity of gastric cancer cells by targeting miR-488-3p / 中国肿瘤临床

Objective: To investigate the miR-488-3p-mediated effects of the long-chain noncoding RNA (lncRNA) family with sequence similarity 201-member A (FAM201A) on the biological behavior and radiosensitivity of gastric cancer cells. Methods: Sixty-three pairs of gastric carcinoma tissues and paracancerous tissues were resected from gastric carcinoma patients, who underwent surgery (no radiotherapy or chemotherapy treatment before surgery) at Laiyang Central Hospital of Yantai City during January 2014 and January 2017. The expression levels of FAM201A and miR-488-3p in the 63 gastric cancer tissue samples and their paracancerous tissues were detected by real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The gastric cancer cell line MGC803 with inhibited FAM201A expression was constructed. The proliferation and viability of MGC803 cells were assessed by the Methyl Thiazolyl Tetrazolium (MTT) assay; their apoptosis was measured by flow cytometry; and their migration and invasion abilities were tested by the Transwell assay. Further, the MGC803 cells were irradiated with different radiation intensities (0, 2, 4, 6, and 8 Gy). Cell survival fractions were measured by colony-formation assay, and cell survival curve was stimulated by the single-hit multi-target model. The dual-luciferase reporter gene assay and qRT-PCR were used to verify whether FAM201A targets miR-488-3p. Results: The expression level of FAM201A was significantly higher, while that of miR-488-3p was significantly lower in gastric cancer tissues than in the paracancerous tissues. Inhibiting FAM201A expression significantly suppressed the proliferation, migration, and invasion abilities of MGC803 cells, promoted their apoptosis, and increased their radiosensitivity. FAM201A negatively regulated miR-488-3p expression. Inhibiting miR-488-3p expression reversed the effects of the inhibition of FAM201A expression on the proliferation, apoptosis, migration, and radiosensitivity of MGC803 cells. Conclusions: Inhibiting the expression of the lncRNA FAM201A suppressed the proliferation, migration, and invasion abilities of gastric cancer cells, promoted their apoptosis, and increased their radiosensitivity by targeting miR-488-3p. Thus, FAM201A may serve as a novel molecular target for gastric cancer treatment.

Long noncoding RNAs are involved in osteoblast differentiation and osteoclast production / 中国组织工程研究

BACKGROUND: Long noncoding RNAs play a role In transcription and post-transcrlptlonal levels, participate in the regulation of bone regeneration, and are closely related to osteoarthritis as well.  OBJECTIVE: To review the research progress of long noncoding RNAs in bone regeneration and osteoarthritis. METHODS: A computer-based search of CNKI, PubMed, and Elsevier was performed for relevant articles regarding long noncoding RNAs in bone regeneration and osteoarthritis published from January 2000 to June 2019, including review, basic research and clinical research. The search terms were “LncRNA; bone; bone regeneration; osteoarthritis” in Chinese and English. After preliminarily reading titles and abstracts, irrelevant articles were excluded. According to the inclusion and exclusion criteria, 64 articles were finally included in result analysis. RESULTS AND CONCLUSION: Bone regeneration is a complex process involving the interaction of osteoblasts and osteoclasts. Long noncoding RNAs are involved in the differentiation of osteoblasts and the production of osteoclasts, which play an important role in the balance of osteoblasts and osteoclasts. Therefore, it is a key molecule of bone regeneration. Long noncoding RNAs are involved in the occurrence and development of osteoarthritis and differentially expressed in osteoarthritis patients and healthy people, some of which are elevated and some are decreased in osteoarthritis patients. Thus, defining their respective expression differences and functions is of great significance for the prevention and treatment of osteoarthritis.

Regulatory mechanism of long noncoding RNA in the occurrence and development of leukemia: a review / 生物工程学报

Long noncoding RNAs (lncRNAs) are a class of RNA molecules that are greater than 200 nt in length and do not have protein-coding capabilities or encode micropeptides only. LncRNAs are involved in the regulation of cell proliferation, differentiation, apoptosis and other biological processes, and are closely associated with the occurrence, recurrence and metastasis of a variety of malignant hematologic diseases. This article summarizes the function, regulatory mechanism and potential clinical application of lncRNAs in leukemia. In general, lncRNAs regulate the occurrence and development of leukemia and the multi-drug resistance in chemotherapy through epigenetic modification, ribosomal RNA transcription, competitive binding with miRNA, modulating glucose metabolic pathway, and activating tumor-related signaling pathway. Studies on lncRNAs provide new references for understanding the pathogenesis of leukemia, uncovering new prognostic markers and potential therapeutic targets, and addressing the problems of drug resistance and post-treatment recurrence in patients in clinical treatment of leukemia.

Overexpression of lncRNA MEG3 inhibits proliferation and invasion of glioblastoma U251 cells / 南方医科大学学报

OBJECTIVE@#To investigate the effects of overexpression of long noncoding RNA (lncRNA) MEG3 on the proliferation and invasion of glioblastoma U251 cells by suppressing the expression of hypoxia inducible factor 1@*METHODS@#The expression of lncRNA MEG3 and HIF1@*RESULTS@#The expression of MEG3 was significantly lower and HIF1@*CONCLUSIONS@#MEG3 overexpression inhibits the proliferation and invasion of U251 cells through suppressing the expression of HIF1

Effect and Mechanism of Xiangsha Liujunzi Tang on Lipid Deposition in Liver of ApoE-/- AS Mice by Affecting Long Noncoding RNA-HC/miR-130b and to Regulate Cholesterol Metabolism / 中国实验方剂学杂志

Objective:To investigate the mechanism of Xiangsha Liujunzi Tang in improving liver lipid deposition in ApoE^-/- atherosclerotic （AS） mice by affecting long noncoding RNA-HC （Lnc-HC）/microRNA-130b （miR-130b） in the regulation of cholesterol metabolism. Method:Totolly 10 C57BL/6J mice were selected as normal controls， and 30 healthy ApoE^-/- mice fed with high fat diet for 12 weeks were then randomly divided into the model group， Xiangsha Liujunzi Tang group（19.12 g·kg^-1·d^-1） and simvastatin group（2.275 mg·kg^-1·d^-1）， with gavage administration for 4 weeks. The serum lipid level of mice was detected by automatic biochemistry analyzer， and the histopathological changes of liver cells were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect expression of long noncoding RNA-HC， and miR-130b. Real-time PCR and Western blot assay were used to detect gene and protein expression of peroxisome proliferator-activated receptor gamma （PPAR<italic>γ</italic>）， liver X receptor （LXR）， ATP-binding cassette transporters A1 （ABCA1）， ATP-binding cassette transporters G1 （ABCG1）， ATP-binding cassette transporters G5 （ABCG5）， and ATP-binding cassette transporters G8 （ABCG8）. Result:Compared with the normal control group， the mice in the model group showed abnormal blood lipids， larger liver cells， obvious fat vacuoles， significantly increased expression of Lnc-HC， miR-130b in liver， and significantly decreased gene and protein expression of PPAR<italic>γ</italic>， LXR， ABCA1， ABCG1， ABCG5， and ABCG8 in mice liver （<italic>P</italic><0.05，<italic>P</italic><0.01）. Compared with the model group， the abnormal blood lipid levels of the mice in the Xiangsha Liujunzi Tang group and the simvastatin group were improved， and the number of fatty vacuoles of liver cells was significantly reduced， the expression of liver Lnc-HC， miR-130b in Xiangsha Liujunzi Tang group decreased significantly （<italic>P</italic><0.05，<italic>P</italic><0.01）， the gene and protein levels of liver PPAR<italic>γ</italic>， ABCA1， ABCG1， ABCG5， ABCG8 in mice of the Xiangsha Liujunzi Tang group and the simvastatin group showed an upward trend. Among them， the gene and protein expression of LXR protein in the liver of the Xiangsha Liujunzi Tang group was significantly up-regulated （<italic>P</italic><0.05）. Conclusion:Xiangsha Liujunzi Tang may improve the lipid deposition in the liver of ApoE^-/-AS mice by affecting Lnc-HC/miR-130b to regulate the cholesterol metabolism process mediated by PPAR<italic>γ</italic>， thus playing a role in preventing and treating AS.

Long non-coding RNAs: From disease code to drug role

Enormous studies have corroborated that long non-coding RNAs (lncRNAs) extensively participate in crucial physiological processes such as metabolism and immunity, and are closely related to the occurrence and development of tumors, cardiovascular diseases, nervous system disorders, nephropathy, and other diseases. The application of lncRNAs as biomarkers or intervention targets can provide new insights into the diagnosis and treatment of diseases. This paper has focused on the emerging research into lncRNAs as pharmacological targets and has reviewed the transition of lncRNAs from the role of disease coding to acting as drug candidates, including the current status and progress in preclinical research. Cutting-edge strategies for lncRNA modulation have been summarized, including the sources of lncRNA-related drugs, such as genetic technology and small-molecule compounds, and related delivery methods. The current progress of clinical trials of lncRNA-targeting drugs is also discussed. This information will form a latest updated reference for research and development of lncRNA-based drugs.

Research progress of miroRNA in the mechanism of cardiac hypertrophy and its role as new drug targets / 国际生物医学工程杂志

Cardiac hypertrophy is a common physiological or pathological process, and pathological cardiac hypertrophy can lead to heart failure, sudden death, etc. The role of microRNA (miRNA or MIR) in myocardial hypertrophy has gradually attracted public attention. miR-1 plays a certain protective role in the occurrence of cardiac hypertrophy. miR-133 is a key factor in the establishment of mast gene program, which is very important for the development of myocardial hypertrophy. Carvedilol and other drugs can regulate the expression of miR-133. miR-208a plays an important physiological role in the cardiovascular system, and its expression level changes dynamically in a variety of cardiovascular diseases such as cardiac hypertrophy, which is closely related to the progression and prognosis of the disease. The expression of miR-199a is up-regulated in pressure-overload cardiac hypertrophy, and it is found that miR-199a can inhibit autophagy of cardiomyocytes and induce the occurrence of cardiac hypertrophy. miR-200c can protect cardiomyocytes through a variety of pathways. miRNA may become an important biomarker or drug therapeutic target for cardiac hypertrophy. With the deepening of the research on non-coding RNAs including miRNA, its regulation on the occurrence of cardiac hypertrophy and the pathological process of heart failure will be further revealed.

Conservation of gene architecture and domains amidst sequence divergence in the hsrω lncRNA gene across the Drosophila genus: an in silico analysis

The developmentally active and cell-stress responsive hsrx locus in Drosophila melanogaster carries two exons, one omega intron, one short translatable open reading frame (ORFx), long stretch of unique tandem repeats and an overlapping mir-4951 near its 30 end. It produces multiple long noncoding RNAs (lncRNAs) using two transcription start and four termination sites. Earlier cytogenetic studies revealed functional conservation of hsrx in several Drosophila species. However, sequence analysis in three species showed poor conservation for ORFx, tandem repeat and other regions while the 16 nt at 50 and 60 nt at 30 splice junctions of the omega intron, respectively, were found to be ultra-conserved. The present bioinformatic study using the splice-junction landmarks in D. melanogaster hsrx identified orthologues in publicly available 34 Drosophila species genomes. Each orthologue carries a short ORFx, ultra-conserved splice junctions of omega intron, repeat region, conserved 30 -end located at mir-4951, and syntenic neighbours. Multiple copies of conserved nonamer motifs are seen in the tandem repeat region, despite a high variability in the repeat sequences. Intriguingly, only the omega intron sequences in different species show evolutionary relationships matching the general phylogenetic history in the genus. Search in other known insect genomes did not reveal sequence homology although a locus with similar functional properties is suggested in Chironomus and Ceratitis genera. Amidst the high sequence divergence, the conserved organization of exons, ORFx and omega intron in this gene’s proximal part and tandem repeats in distal part across the Drosophila genus is remarkable and possibly reflects functional importance of higher order structure of hsrx lncRNAs and the small omega peptide.

Prognostic value of long non-coding RNA TP73-AS1 expression in different types of cancer: a systematic review and meta-analysis

Background: TP73 antisense RNA 1 (TP73-AS1), a newly discovered long non-coding RNA (lncRNA), has been reported to be upregulated in various kinds of tumors, and shows a variable influence on living quality and prognosis of patients. Thus, we conducted a meta-analysis to evaluate the overall prognostic value of the lncRNA TP73-AS1 in cancer patients. Results: A systematic literature retrieval was carried out using the PubMed, Cochrane Library, EMBASE, and Web of Science databases. We calculated the pooled hazard ratio (HR) and odds ratio (OR) with 95% confidence intervals (CIs) to evaluate the association of TP73-AS1 expression with prognostic and clinicopathological parameters. A total of 15 studies including 1057 cancer patients were finally selected for the meta-analysis. The results demonstrated that high TP73-AS1 expression was significantly associated with shorter overall survival (OS) (HR = 1.97, 95% CI: 1.68­2.31, P b 0.001). According to a fixed-effects or random-effects model, elevated TP73-AS1 expression markedly predicted advanced clinical stage (OR = 3.30, 95% CI: 2.35­4.64, P b 0.001), larger tumor size (OR = 2.37, 95% CI: 1.75­3.22, P b 0.001), earlier lymph node metastasis (OR = 3.28, 95% CI: 1.59­6.76, P = 0.001), and distant metastasis (OR = 4.94, 95% CI: 2.61­9.37, P b 0.001). Conclusions: High lncRNA TP73-AS1 expression appears to be predictive of a worse OS and clinicopathologic features for patients with various types of malignant tumors. These results provide a basis for utilizing TP73- AS1 expression as an unfavorable indicator to predict survival outcomes.