Complementary Sites Research Articles

Long Noncoding RNA FGD5-AS1 Knockdown Decrease Viability, Migration, and Invasion of Non-Small Cell Lung Cancer (NSCLC) Cells by Regulating the MicroRNA-944/MACC1 Axis.

Objective:Long noncoding RNA FGD5 antisense RNA 1 (FGD5-AS1) participates in the regulation of non-small cell lung cancer (NSCLC) progression, but the underlying mechanisms are not fully revealed. This study aimed to determine the regulatory mechanism of FGD5-AS1 on the viability, migration, and invasion of NSCLC cells.Methods:QRT-PCR was performed to measure the expression of FGD5-AS1, microRNA-944 (miR-944), and MACC1 in NSCLC. The correlation between FGD5-AS1 and clinicopathological features of NSCLC patients was analyzed. The viability of NSCLC cells were detected using MTT assay, and the migration and invasion were measured by transwell assay. Additionally, dual-luciferase reporter assay was used to demonstrate the interactions among FGD5-AS1, miR-944, and MACC1. Furthermore, exosomes were isolated from NSCLC cells and identified by transmission electron microscopy (TEM) and western blot. Then, the macrophages treated with exosomes were co-cultured with NSCLC cells to assess the effect of exosomes containing lower FGD5-AS1 level on NSCLC.Results:The expression of FGD5-AS1 and MACC1 was increased in NSCLC, but miR-944 expression was decreased. FGD5-AS1 expression had significantly correlation with TNM stage and metastasis in NSCLC patients. FGD5-AS1 knockdown decreased the viability, migration, and invasion of NSCLC cells. Additionally, FGD5-AS1 and MACC1 were both targeted by miR-944 with the complementary binding sites at 3’ UTR. In the feedback experiments, miR-944 inhibition or MACC1 overexpression reversed the reduction effect of FGD5-AS1 knockdown on the tumorigenesis of NSCLC. Moreover, silencing of FGD5-AS1 suppressed macrophages M2 polarization, and eliminated the promoting effects of exosomes mediated macrophages on NSCLC cell migration and invasion.Conclusions:FGD5-AS1 knockdown attenuated viability, migration, and invasion of NSCLC cells by regulating the miR-944/MACC1 axis, providing a new therapeutic target for NSCLC.

MicroR-1294-5p inhibits glycolytic metabolism of non-small cell lung cancer cells via targeting TMPRSS11B

Non-small cell lung cancer (NSCLC) cells intake and consume glucose at high efficiency by aerobic glycolysis to maintain robust cell growth and resist cell death. MicroRNAs (miRNAs) have been known to play pivotal roles in NSCLC development partly through mediating glycolysis. However, only a few miRNAs have been experimentally confirmed as critical regulators of glycolysis in NSCLC. TCGA datasets were analyzed to screen for differentially expressed miRNAs between NSCLC and normal tissues. The function of miR-1294-5p was determined in NSCLC cells by cell proliferation, glucose uptake, lactate release, and Extracellular Acidification Rate (ECAR) assays. The target of miR- 1294-5p was predicted by TargetScan and miRDB, which was further validated by flow cytometry analysis, RT-qPCR, western blotting, a dual-luciferase reporter assay, and RNA immunoprecipitation (RIP) assay. In the present study, it was found that miR-1294-5p was a significantly downregulated miRNA in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). The overexpression of miR-1294-5p inhibited glycolysis, lactate export, ECAR, and cell proliferation in NSCLC cells. Analysis with bioinformatic tools, Western Blotting, RT-qPCR, flow cytometry analysis, dual-luciferase reporter assay, and RIP assay showed that miR-1294-5p directly bound to complementary sites in the 3’-Untranslated Region (UTR) of TMPRSS11B resulted in downregulation of TMPRSS11B expression. In addition, transfection of recombinant TMPRSS11B rescued the functions of miR-1294-5p on glycolysis and proliferation of NSCLC cells. The findings provided novel insights for understanding the regulation of glycolytic metabolism in NSCLC.

The proposal of a GSSP for the Berriasian Stage (Cretaceous System): Part 2

In part 1 of this work we discussed the possibilities for the selection of a GSSP for the Berriasian Stage of the Cretaceous System, based on prevailing practical methods for correlation in that J/K interval, traditional usage and the consensus over the best boundary markers that had developed in the last forty years. This consensus has developed further, based on the results of multidisciplinary studies on numerous sites over the last decade. Here in Part 2 we give an account of the application of those results by the Berriasian Working Group (ISCS), and present the stratigraphic evidence that justifies the selection of the locality of Tré Maroua (Hautes-Alpes, SE France) as the proposed GSSP. We describe a 45 m-thick section in the Calcaires Blancs vocontiens – that part of the formation covering the calpio­nellid Chitinoidella, Remanei. Intermedia, Colomi, Alpina, Ferasini, Elliptica and Simplex biozones. The stratigraphic data collected here has been compiled as part of a wider comparative study of complementary Vocontian Basin sites (with localities at Charens, St Bertrand, Belvedere and Le Chouet). Evidence from Tré Maroua thus sits in this substantial regional biostratigraphic and magnetostratigraphic context. For the purposes of the GSSP definition, here we particularly concentrate on the unbroken sequence and biotic markers in the interval immediately below the boundary, the Colomi Subzone (covering circa 675,000 years), and immediately above, the Alpina Subzone (covering circa 725,000 years). Particularly significant fossil datums identified in the Tré Maroua profile are the primary basal Berriasian marker, the base of the Alpina Subzone (a widespread event marked by dominance of small Calpionella alpina, with rare Crassicollaria parvula and Tintinopsella carpathica): the base of the Berriasian Stage is placed at the base of bed 14, which coincides with the base of the Alpina Subzone. Secondary markers bracketing the base of the Calpionella Zone are the FOs of the calcareous nannofossil species Nannoconus wintereri, close below the boundary, and the FO of Nannoconus steinmannii minor, close above. The Tithonian/Berriasian boundary level occurs within M19n.2n, in common with many documented sites, and is just below the distinctive reversed magnetic subzone M19n.1r (the so-called Brodno reversal). We present data which is congruent with magnetostratigraphic and biostratigraphic data from other key localities in France and in wider regions (Le Chouet, Saint Bertrand, Puerto Escaño, Rio Argos, Bosso, Brodno, Kurovice, Theodosia…), and thus the characteristics and datums identified at Tré Maroua are key for correlation and, in general, they typify the J/K boundary interval in Tethys and connected seas.

Regioselective Functionalization of Ester-, Amide-, Carbonate-, and Carbamate-Substituted 2-Phenyl-2-oxazolines with Mixed Lithium-Magnesium Amides.

We have prepared novel highly functionalized benzene derivatives by regioselective metalation of ester-, amide-, carbamate-, and carbonate-substituted 2-phenyl-2-oxazolines with mixed lithium-magnesium amides followed by reaction with different electrophiles. While a complementary metalation site can be accessed by using different bases, steric and electronic effects promoted by the aromatic ring substituents also play an important role in reaction regioselectivity. Computational calculations of the aromatic hydrogen pKa values have helped us to rationalize the metalation preference by the complex-induced proximity effect concept.

MiR-409-3p inhibits the proliferation and migration of human ovarian cancer cells by targeting Rab10

Ovarian cancer is a leading cause of gynecological cancer-related mortality. It has been reported that miR-409-3p is involved in the proliferation and migration of cancer cells. However, the role of miR-409-3p in ovarian cancer has not been well studied. The present study aimed to investigate the functional role of miR-409-3p in the pathogenesis of ovarian cancer, and its potential mechanism. It was found that the expression levels of miR-409-3p in 6 ovarian cancer tissues were upregulated. Through proliferation, migration and colony formation assays, it was revealed that overexpression of miR-409-3p inhibited the proliferation and migration of ovarian cancer cells. It was predicted from bioinformatics assays that the complementary binding sites were within miR-409-3p and Rab10. It was also demonstrated that the downregulation of the expression of Rab10 reversed the miR-409-3p downregulation-induced abnormal proliferation of ovarian cancer cells. These results suggest that miR-409-3p expression can be used as a predictive marker for the prognosis of ovarian cancer. Thus, the miR-409-3p/Rab10 axis may be a novel therapeutic target for ovarian cancer.

LncRNA LINC01535 upregulates BMP2 expression levels to promote osteogenic differentiation via sponging miR‑3619‑5p.

Osteoporosis is a debilitating skeletal disease that causes bones to collapse and is accompanied by a high risk of bone fracture. It was previously demonstrated that the osteogenic differentiation of human bone marrow‑derived mesenchymal stem cells (hBMSCs) serves an important role in the process of human bone formation. Accumulating research has indicated that long non‑coding RNAs (lncRNAs) participate in hBMSC osteogenic differentiation. For example, LINC01535 was reported to serve as a carcinogenic factor in cervical cancer; however, its latent function and molecular mechanism in the osteogenesis of hBMSCs remain to be investigated. The present study showed that the expression levels of LINC01535 were upregulated upon increasing osteogenic differentiation time. In addition, the inhibition of LINC01535 inhibited hBMSC proliferation and osteogenic differentiation and promoted cell apoptosis. Using bioinformatics analysis, LINC01535 was discovered to have complementary binding sites for microRNA (miR)‑3619‑5p, and further experiments demonstrated that LINC01535 functioned as a sponge of miR‑3619‑5p. Additionally, bone morphogenetic protein2 (BMP2) was confirmed to be a target of miR‑3619‑5p. The results revealed that LINC01535 regulated the expression levels of BMP2 via sponging miR‑3619‑5p. In conclusion, the findings of the present study suggested that LINC01535 may accelerate the osteogenic process of hBMSCs via targeting the miR‑3619‑5p/BMP2 axis, which may offer an innovative therapeutic method for osteoporosis.

Down-regulation of Exosomal miR-214-3p Targeting CCN2 Contributes to Endometriosis Fibrosis and the Role of Exosomes in the Horizontal Transfer of miR-214-3p.

Endometriosis (EMs) is defined as the presence of tissue which somewhat resembles endometrial glands and stroma outside the uterus, and elicits fibrosis. Fibrosis is the main factor resulting in pain and infertility, while the aetiology of endometrial fibrosis is unknown. There is strong evidence from numerous experiments showing that connective tissue growth factor (CCN2) plays a central role in fibrogenesis. Exosomal miR-214-3p can regulate the expression of CCN2 through binding to complementary sites in the 3' untranslated region. This study aimed to explore the role of exosomal miR-214-3p in endometriosis fibrosis and the relationship between CCN2 and miR-214-3p in endometriosis fibrosis. Our results demonstrated that miR-214-3p was significantly down-regulated and CCN2 was up-regulated in EMs ectopic lesion and stromal cells compared with EMs eutopic and endometrium of patients without endometriosis. Exosomal miR-214-3p can inhibit fibrosis in EMs through targeting CCN2. The results were explored and verified in vitro and in vivo, respectively. Cell co-culture was used to explore the contributions of exosomes to intercellular information transmission of miR-214-3p. The results showed that exosomes play a pivotal role in the transportation of miR-214-3p between cells. Furthermore, level of exosomal miR-214-3p in endometriosis patients' serum was lower than that in patients without endometriosis. In conclusion, exosomal miR-214-3p can inhibit fibrosis in EMs by targeting CCN2. MiR-214-3p may be considered as a bio-marker and has a potential therapeutic effect in EMs.

Why forget us? Developing the Three-Town enslaved relics as alternative sites of commemoration in southeastern Ghana

ABSTRACT This paper examines preliminary research findings emanating from investigations carried out on the marginalised landscapes and relics of slavery in the Three-Town area of southeastern Ghana. It discusses the politics of exclusion being practised in the choice of histories and sites of enslavement for annual commemorative events in Ghana. The popular knowledge that the trade in humans occurred mostly at Cape Coast and Elmina Castles in the Central Region of Ghana has skewed development and commemorative activities such as Emancipation, PANAFEST and Year of Return programmes toward these exclusive sites of reverence. Lack of attention to the Three-Town sites and other sites of enslavement and their relics by government institutions has exacerbated their exclusion from public discourse and heritage development. This paper questions this indifference by highlighting the tangible and intangible imprints of trans-Atlantic slavery and enslavement on the histories, landscapes and memories of the Somey-Ewe people of Three-Town. It underscores the need — and the ongoing quest by archaeologists — to develop the Three-Town enslaved relics as important enslaved heritage resources and complementary sites of reverence in southeastern Ghana.

Long noncoding RNA LINC00858 promotes the proliferation, migration and invasion of gastric cancer cells via the miR-363-3p/FOXP4 axis.

Long non-coding RNA (lncRNA) LINC00858 has been found to exert oncogenic activity in several types of cancers, except gastric cancer (GC). In the present study, we aimed to explore the potential role of LINC00858 in GC and the underlying molecular mechanism. The expression patterns of LINC00858 were determined using qRT-PCR in GC samples and cell lines. Cell proliferation was examined utilizing CCK-8 assay. Cell migration and invasion were evaluated using transwell assays. We used the bioinformatics software StarBase and TargetScan to predict lncRNA-miRNA and miRNA-mRNA interactions. Our findings revealed that LINC00858 expression was markedly upregulated in GC tissues and cell lines. Loss-of-function experiments demonstrated that LINC00858 silencing inhibited the proliferation, migration and invasion of GC cells. Bioinformatics analysis showed that there were several complementary binding sites between LINC00858 and microRNA (miR)-363-3p, and further Luciferase reporter assay confirmed the interaction between LINC00858 and miR-363-3p. In addition, forkhead box P4 protein (FOXP4) was found to be a target gene of miR-363-3p in GC cells. FOXP4 overexpression reversed the inhibitory effects of miR-363-3p mimics on cell proliferation, migration and invasion of BGC-823 cells. Collectively, LINC00858 acted as an oncogene in GC via regulating miR-363-3p/FOXP4 axis, which indicated that LINC00858 might be a novel therapeutic target for the treatment of GC.

Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction.

MicroRNAs (miRNAs) are single stranded, non-coding RNA molecules that typically regulate gene expression at the post-transcriptional level by binding to partially complementary target sites in the 3' untranslated region (UTR) of messenger RNA (mRNA), which reduces the mRNA's translation and stability. The miRNA expression profiles in various organs and tissues of mice have been investigated, but standard methods for the purification and quantification of miRNA in mouse kidney have not been available. We have established an effective and reliable method for extracting and evaluating miRNA expression in mouse kidney with renal interstitial fibrosis by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The protocol required five steps: (1) creation of sham and unilateral ureteral obstruction (UUO) mice; (2) extraction of kidney samples from the UUO mice; (3) extraction of total RNA, which includes miRNA, from the kidney samples; (4) complementary DNA (cDNA) synthesis with reverse transcription from miRNA; and (5) qRT-PCR using the cDNA. Using this protocol, we successfully confirmed that compared to the controls, the expression of miRNA-3070-3p was significantly increased and those of miRNA-7218-5p and miRNA-7219-5p were significantly decreased in the kidneys of a mouse model of renal interstitial fibrosis. This protocol can be used to determine the miRNA expression in the kidneys of mice with UUO.

CRISPR GUARD protects off-target sites from Cas9 nuclease activity using short guide RNAs

Precise genome editing using CRISPR-Cas9 is a promising therapeutic avenue for genetic diseases, although off-target editing remains a significant safety concern. Guide RNAs shorter than 16 nucleotides in length effectively recruit Cas9 to complementary sites in the genome but do not permit Cas9 nuclease activity. Here we describe CRISPR Guide RNA Assisted Reduction of Damage (CRISPR GUARD) as a method for protecting off-targets sites by co-delivery of short guide RNAs directed against off-target loci by competition with the on-target guide RNA. CRISPR GUARD reduces off-target mutagenesis while retaining on-target editing efficiencies with Cas9 and base editor. However, we discover that short guide RNAs can also support base editing if they contain cytosines within the deaminase activity window. We explore design rules and the universality of this method through in vitro studies and high-throughput screening, revealing CRISPR GUARD as a rapidly implementable strategy to improve the specificity of genome editing for most genomic loci. Finally, we create an online tool for CRISPR GUARD design.

Long-Chain Non-Coding RNAs Metastasis-Associated Lung Adenocarcinoma Transcript-1 Is Involved in Growth of Nasopharyngeal Carcinoma Cells by Targeting miR-205-5p

The incidence of nasopharyngeal carcinoma (NC) has been rising. The prognosis of NC remains unsatisfactory; therefore, it is necessary to find new ways to improve it. LncRNA MALAT1 (MALAT1) is a well-studied gene, but its status in NC remains unclear. More experimental analyses are needed to uncover the role of MALAT1 in NC for a reliable and accurate theoretical basis for NC’s diagnosis and treatment. First, NC patients admitted to our hospital from February 2018 to March 2020 and healthy controls who underwent physical examinations during the same period were enrolled in this study. MALAT1 and miR-205-5p in the patients’ peripheral blood and tissues were detected. The expression of MALAT1 was high, and the expression of miR-205-5p was low in the NC patients. Both genes were effective in predicting the occurrence of NC, and their expression was negatively correlated. According to in vitro experiments, inhibiting MALAT1 and increasing miR-205-5p could reduce CNE-2Z cells’ proliferation and increase their apoptotic rate. However, increasing MALAT1 and inhibiting miR-205-5p had opposite effects. Through the online website ENCORI, complementary sites were found in MALAT1 and miR-205-5p. According to dual-luciferase reporter gene assay, MALAT1-WT’s luciferase activity was inhibited by the co-transfection of miR-205-5p-min, while MALAT1-MUT’s luciferase activity was enhanced by the co-transfection of miR-205-5p-inh. Lastly, through rescue experiments, we found no difference in the biological behavior between the cells in the MALAT1 si-RNA+miR-205-5p-inh group and the MALAT1+NC-RNA group. Therefore, MALAT1 promotes NC cell proliferation, inhibits apoptosis, and participates in NC’s development via specifically regulating miR-205-5p.

Suppression of circular RNA circDHCR24 alleviates aortic smooth muscle cell proliferation and migration by targeting miR-149-5p/MMP9 axis

Circular RNAs (circRNAs) are involved in many courses of atherosclerosis and coronary artery disease (CHD). However, the role and effect of circRNAs in vascular restenosis after PCI remains unclear. Human aortic vascular smooth muscle cell (HA-VSMC) was cultured and stimulated with PDGF-BB. The expression profile of circRNAs in HA-VSMCs was screened using microarray analysis. A total 257 aberrantly expressed circRNAs were screened with 2 fold change. Has_circ_0113656 (also called circDHCR24) was validated by qRT-PCR to be significantly up-regulated in PDGF-BB induced HA-VSMCs. CircDHCR24 silencing obviously inhibited the proliferation, migration and phenotypic switch. Moreover, bioinformatics analysis predicted that miR-149-5p had complementary binding sites in 3′-UTR of circDHCR24. Luciferase reporter assay and RIP assay further verified the circDHCR24 acts as a spong for miR-149-5p in HA-VSMCs. Besides, bioinformatics analysis, luciferase reporter assay and RIP assay proved MMP9 was a directly target of miR-149-5p. Finally, cells were transfected with si-circDHCR24 with or without miR-149 inhibitor, and the results showed that co-transfection si-circDHCR24 and miR-149 inhibitor reversed the effect of si-circDHCR24 on cell proliferation, migration and phenotypic switch. Taken together, our study suggested for the first time that the knockdown of circDHCR24 alleviates HA-VSMCs proliferation, migration and phenotypic switching, thereby preventing vascular restenosis.

LEMD1-AS1 Suppresses Ovarian Cancer Progression Through Regulating miR-183-5p/TP53 Axis.

Background/AimsLong noncoding RNAs (lncRNAs) play a critical role in tumorigenesis and progression of ovarian cancer (OC). This study focused on the function and potential mechanism toward LEMD1-AS1 (LEMD1 antisense RNA 1) in the progression of ovarian cancer.Materials and MethodsThe expression of LEMD1-AS1 in OC tissues was evaluated in TCGA and Gene Expression Omnibus datasets (GSE119056) and confirmed in OC cell lines via qRT-PCR (quantitative real-time polymerase chain reaction). Then, the location of LEMD1-AS1 in the cytoplasmic and nuclear RNAs extracted from OV cells was detected by qRT-PCR. Cell Counting Kit-8 (CCK-8), colony formation, wound-healing and transwell assays were applied to examine cell viability, proliferation, migration and invasion, respectively. Further, the effect of LEMD1-AS1 on OC tumor growth was determined via subcutaneous xenotransplanted tumor model. The potential target for LEMD1-AS1 was validated via dual-luciferase activity assay, RNA pull-down and RNA immunoprecipitation.ResultsThe expression of LEMD1-AS1 was decreased in OC tissues and cell lines. Forced overexpression of LEMD1-AS1 inhibited the proliferation, migration and invasion of ovarian cancer cells and transplanted tumor growth in nude mice. We found that LEMD1-AS1 was mainly located in the cytoplasm of OC cells and contained complementary sites of miR-183-5p. Mechanistically, our results showed that LEMD1-AS1 could directly interact with miR-183-5p and tumor protein p53 (TP53). The anti-tumor role of LEMD1-AS1 on OC progression depended on miR-183-5p-mediated TP53 expression.ConclusionLEMD1-AS1 suppresses OC progression through sponging miR-183-5p and regulation of TP53, suggesting a novel biomarker and target for OC.

A Review of MicroRNA in Uveal Melanoma.

Uveal melanoma (UM) is the most common and aggressive primary intraocular tumor in adults. UM is classified as a malignant tumor with a strong tendency of metastasis, which always leads to poor outcomes. At present, the pathogenesis of UM remains unclear and lacks effective therapies. Recent studies have shown that microRNAs (miRNAs), defined as a group of 21–23 nucleotides single-stranded noncoding RNAs, play a significant role in UM. By binding to the complementary sites within the 3ʹ untranslated region (3ʹUTR) of message RNAs (mRNAs), miRNAs regulate genes by decaying mRNAs or inhibiting their translation. Thus, miRNAs can modulate various biological behaviors of tumors, including cell proliferation, invasion and metastasis. Furthermore, miRNAs have shown clinical applications by serving as biomarkers for diagnosis and prognosis, regulating immune response, and functioning as epigenetic regulators. It is reasonable to believe that miRNAs have wide application prospects in the early diagnosis and therapy of UM.

Encoding Multiple Reactivity Modes within a Single Synthetic Replicator.

Establishing programmable and self-sustaining replication networks in pools of chemical reagents is a key challenge in systems chemistry. Self-replicating templates are formed from two constituent components with complementary recognition and reactive sites via a slow bimolecular pathway and a fast template-directed pathway. Here, we re-engineer one of the components of a synthetic replicator to encode an additional recognition function, permitting the assembly of a binary complex between the components that mediates replicator formation through a template-independent pathway, which achieves maximum rate acceleration at early time points in the replication process. The complementarity between recognition sites creates a key conformational equilibrium between the catalytically inert product, formed via the template-independent pathway, and the catalytically active replicator that mediates the template-directed pathway. Consequently, the rapid formation of the catalytically inert isomer kick-starts replication through the template-directed pathway. Through kinetic analyses, we demonstrate that the presence of the two recognition-mediated reactivity modes results in enhanced template formation in comparison to that of systems capable of exploiting only a single recognition-mediated pathway. Finally, kinetic simulations reveal that the conformational equilibrium and both the relative and absolute efficiencies of the recognition-mediated pathways affect the extent to which self-replicating systems can benefit from this additional template-independent reactivity mode. These results allow us to formulate the rules that govern the coupling of replication processes to alternative recognition-mediated reactivity modes. The interplay between template-directed and template-independent pathways for replicator formation has significant relevance to ongoing efforts to design programmable and adaptable replicator networks.

Influence of the variability of soil profile properties on weak and strong seismic response

Characterizing the potential effect of local site conditions on the amplification of ground motions is a critical aspect of seismic hazard and risk assessment. The aim of this study is to investigate the reliability and the limit of using the average shear wave velocity in the upper 30m of the soil profile vs,30, as single proxy, to characterize seismic site effects for weak and strong events.To this regard, a dataset of 300 one-dimensional soil profiles with a given vs,30 are generated through a Monte Carlo approach. Their seismic responses are computed for a set of 40 real accelerograms, with different seismic features. The vertical propagation from the bottom of the generated columns is modeled using a finite element spatial discretization, accounting for both linear and nonlinear soil behavior.The site dominant frequency f0 and the shear wave velocity gradient in the profile B30 are proposed as proxies to characterize seismic site effects and the variability of the response spectra for the numerical signals, at the free surface of the set of columns, is discussed. Correlations between site-specific amplification factors deduced using the numerical response spectra and the proposed site proxies are analyzed for different sub-ranges of periods. The obtained amplification factors are then compared to those proposed by different international and national design codes. The results, obtained under assumption of linear and nonlinear behavior of soil, emphasize the need to introduce complementary site parameters proxies, in addition to vs,30, to characterize the expected site effects in design response spectra.

E2F1-Induced Overexpression of Long Noncoding RNA SBF2-AS1 Promotes Non-Small-Cell Lung Cancer Metastasis Through Regulating miR-362-3p/GRB2 Axis.

Long noncoding RNA SBF2 antisense RNA 1 (lncRNA SBF2-AS1) has been reported to be involved in non-small-cell lung cancer (NSCLC) tumorigenesis. However, the biological role and regulatory mechanism of lncRNA SBF2-AS1 on NSCLC metastasis remain largely unknown. In this study, the expression level and functional role of SBF2-AS1 were investigated in both NSCLC tissues and cell lines. We found that SBF2-AS1 was upregulated in both NSCLC tissues and cell lines. Patients with high levels of SBF2-AS1 have larger tumors, higher malignancy, and poor prognosis. Knockdown of SBF2-AS1 significantly inhibited tumor growth in vivo and cell proliferation, migration, and invasion in vitro. Moreover, bioinformatics analysis, chromatin immunoprecipitation assay, and luciferase reporter assay proved that the upregulation of SBF2-AS1 was mediated by transcription factor E2F1. Further experiments demonstrated that miR-362-3p had complementary binding site with 3'-UTR of SBF2-AS1. Besides, luciferase reporter assay validated that GRB2 was the target protein of miR-362-3p. Rescue experiments showed that SBF2-AS1 silencing inhibited cell invasion and migration, while cotransfection si-SBF2-AS1 and miR-362-3p inhibitor rescued the effect of si-SBF2-AS1. These results demonstrate that E2F1-induced overexpression of SBF2-AS1 promotes the expression of GRB2 by targeting miR-362-3p to facilitate the metastasis of NSCLC.

Genetic Similarity of two different color Local Guinea Fowl (Numida Meleagris) By Using RAPD-PCR Technique

The aim of present study to determine DNA polymorphism, between and within two different color (white) and (black) guinea fowl lines. Blood samples were collected about 3 ml of 30 white guinea fowl and 30 black guinea fowl, in six different local farms. DNA was isolated, the purity of the DNA samples ranged from 1.8 to 1.9. RAPD – PCR was used to evaluate genetic similarity between the lines of guinea fowl. Using eighteen primers for amplification of RAPD, from Gen Script USA. A total of (14) primers out of (18) primers gave results to find Genomic DNA complementary sites. After electrophoresis the largest number of bands were appeared with OPA-03 primers, but the smallest was obtained when OPQ-15 primer used. Determined Genetic similarity as the mean for the primers used, between the black and white guinea fowl, was 0.76. The coefficient of genetic similarity within white guinea fowl line, was 0.78 and 0.73 within black guinea fowl line.

PD24-06 MOLECULAR ALTERATION OF GABAERGIC PAIN MODULATORY PATHWAY IN THE ROSTRAL VENTROMEDIAL MEDULLA FOLLOWING ZYMOSAN-INDUCED CYSTITIS IN RATS

You have accessJournal of UrologyBladder & Urethra: Anatomy, Physiology & Pharmacology II (PD24)1 Apr 2020PD24-06 MOLECULAR ALTERATION OF GABAERGIC PAIN MODULATORY PATHWAY IN THE ROSTRAL VENTROMEDIAL MEDULLA FOLLOWING ZYMOSAN-INDUCED CYSTITIS IN RATS Bidyut K. Medda*, Sankar Addya, Bhavana Talluri, Faith Hoelzel, Maia Terashvili, Patrick Sanvanson, Reza Shaker, Jyoti N. Sengupta, and Banani Banerjee Bidyut K. Medda*Bidyut K. Medda* More articles by this author , Sankar AddyaSankar Addya More articles by this author , Bhavana TalluriBhavana Talluri More articles by this author , Faith HoelzelFaith Hoelzel More articles by this author , Maia TerashviliMaia Terashvili More articles by this author , Patrick SanvansonPatrick Sanvanson More articles by this author , Reza ShakerReza Shaker More articles by this author , Jyoti N. SenguptaJyoti N. Sengupta More articles by this author , and Banani BanerjeeBanani Banerjee More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000881.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The rostral ventromedial medulla (RVM) comprises of several subsets of neurons, which send descending projections to the spinal cord to modulate pain. These neurons are the major targets of efferent projections from the periaqueductal gray (PAG), the site of action of opioid-mediated analgesia. The majority of RVM originating descending neurons are GABAergic. Recently, we have documented microRNA (miRNA)-mediated transcriptional deregulation of spinal GABAergic system and visceral hypersensitivity (colon and bladder) in rats with zymosan-induced cystitis. The objective was to study the effect of noxious bladder insult on molecular expression of GABAergic RVM neurons of rats with zymosan-induced cystitis. METHODS: Two groups of Sprague Dawley rats (n=8 /group) were used in this study. Experimental and control groups received transurethral zymosan (1% in saline, 300ul) and saline respectively for 3 days from postnatal (P) days P58 through P60. Rats in both groups were euthanized on P60 and RVM tissues were collected for analysis. Gene expression profiles of experimental and control groups were evaluated using Rat Clariton S transcriptome arrays. miRNA expression profiles were examine using Affymatrix Gene Chips miRNA arrays. Individual gene was further evaluated using immunohistochemistry (IHC) and in situ hybridization (ISH). RESULTS: Intravesical zymosan treatment resulted in 4-fold downregulation of GABAA receptor subunit alpha 6 (GABAAα6) and glycine transporter Gly 2 (p<0.05 vs controls). Fourteen miRNAs demonstrated differential expression in zymosan treated rats (p<0.05 vs controls). The target gene prediction identified two upregulated miRNAs; miR-129-5p and miR-300 with complementary binding sites for 3’UTRs of GABAAα6 gene indicating a possible post-transcriptional deregulation of the RVM GABAergic pathway in zymosan-treated cystitis rats. A significant downregulation of RVM vesicular GABA transporter VGAT expression was observed in cystitis group by ISH (p<0.05 vs controls). The expression profile of GABAAα6 in RVM was validated by IHC. CONCLUSIONS: Cystitis results in miRNA-mediated transcriptional deregulation of GABA receptor and transporter expression in RVM neurons. This finding strongly suggests that the suppression of the RVM inhibitory transmission may contribute towards the development of spinal neuron hyperexcitability and bladder pain. Source of Funding: NIH R01DK099201 © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e534-e534 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Bidyut K. Medda* More articles by this author Sankar Addya More articles by this author Bhavana Talluri More articles by this author Faith Hoelzel More articles by this author Maia Terashvili More articles by this author Patrick Sanvanson More articles by this author Reza Shaker More articles by this author Jyoti N. Sengupta More articles by this author Banani Banerjee More articles by this author Expand All Advertisement PDF downloadLoading ...