Urothelial Cancer Associated 1 Research Articles

The Role of Urothelial Cancer-Associated 1 in Gynecological Cancers.

Gynecological cancers (GC) represent some of the most frequently diagnosed malignancies in women worldwide. Long-non-coding RNAs (lncRNAs) are regulatory RNAs increasingly being recognized for their role in tumor progression and metastasis in various cancers. Urothelial cancer-associated 1 (UCA1) is a lncRNA, first found deregulated in bladder cancer, and many studies have exposed its oncogenic effects in more tumors since. However, the role of UCA1 in gynecological malignancies is still unclear. This review aims to analyze and define the role of UCA1 in GC, in order to identify its potential use as a diagnostic, prognostic, or therapeutic biomarker of GC. By employing the search terms "UCA1", "breast cancer", "endometrial cancer", "ovarian cancer", "cervical cancer", "vaginal cancer", and "vulvar cancer" in the PubMed database for the literature review, we identified a total of sixty-three relevant research articles published between 2014 and 2024. Although there were some opposing results, UCA1 was predominantly found to be upregulated in most of the breast, endometrial, ovarian, cervical, and vulvar cancer cells, tissue samples, and mouse xenograft models. UCA1 overexpression mainly accounts for enhanced tumor proliferation and increased drug resistance, while also being associated with some clinicopathological features, such as a high histological grade or poor prognosis. Nonetheless, no reviews were identified about the involvement of UCA1 in vaginal carcinogenesis. Therefore, further clinical trials are required to explore the role of UCA1 in these malignancies and, additionally, examine its possible application as a target for upcoming treatments, or as a novel biomarker for GC diagnosis and prognosis.

Endogenous retrovirus HERVH-derived lncRNA UCA1 controls human trophoblast development

Endogenous retroviruses (ERVs) are frequently reactivated in mammalian placenta. It has been proposed that ERVs contribute to shaping the gene regulatory network of mammalian trophoblasts, dominantly acting as species- and placental-specific enhancers. However, whether and how ERVs control human trophoblast development through alternative pathways remains poorly understood. Besides the well-recognized function of human endogenous retrovirus-H (HERVH) in maintaining pluripotency of early human epiblast, here we present a unique role of HERVH on trophoblast lineage development. We found that the LTR7C/HERVH subfamily exhibits an accessible chromatin state in the human trophoblast lineage. Particularly, the LTR7C/HERVH-derived Urothelial Cancer Associated 1 (UCA1), a primate-specific long non-coding RNA (lncRNA), is transcribed in human trophoblasts and promotes the proliferation of human trophoblast stem cells (hTSCs), whereas its ectopic expression compromises human trophoblast syncytialization coinciding with increased interferon signaling pathway. Importantly, UCA1 upregulation is detectable in placental samples from early-onset preeclampsia (EO-PE) patients and the transcriptome of EO-PE placenta exhibits considerable similarities to that of the syncytiotrophoblasts differentiated from UCA1-overexpressing hTSCs, supporting up-regulated UCA1 as a potential biomarker of this disease. Altogether, our data shed light on the versatile regulatory role of HERVH in early human development and provide a unique mechanism whereby ERVs exert a function in human placentation and placental syndromes.

LncRNA urothelial cancer associated 1 promotes the osteogenic differentiation of human periodontal ligament stem cells by regulating the miR-96–5p/Osx axis

ObjectivesTo investigate the expression of long non-coding RNA (lncRNA) urothelial cancer associated 1 (UCA1) in human periodontal ligament stem cells (hPDLSCs), its effect on osteogenic differentiation of hPDLSCs and its mechanism. DesignThe expression of osteogenic genes Osx, Runx2, Ocn and Opn was explored by qPCR. Protein expression in hPDLSCs was estimated by Western blot. The osteogenic differentiation of hPDLSCs was detected by Alizarin red staining assays. The interaction between UCA1 and miR-96–5p was explored by RNA pulldown assay and dual luciferase assay. The interaction between miR-96–5p and Osx 3′-UTR was measured by dual luciferase assay. ResultsThe expression of UCA1 and miR-96–5p was negatively correlated in hPDLSCs. During the osteogenic differentiation of hPDLSCs, the expression of UCA1 was increased, while the expression of miR-96–5p was decreased. Knockdown of UCA1 in hPDLSCs inhibited osteogenic differentiation but induced upregulation of miR-96–5p expression, and vice versa. In addition, miR-96–5p partially reversed the positive effect of UCA1 on osteogenic differentiation of hPDLSCs. Notably, UCA1 was identified as a miR-96–5p sponge, and miR-96–5p targeted Osx. ConclusionsOur results demonstrated that the novel UCA1/miR-96–5p/Osx pathway regulates osteogenic differentiation of hPDLSCs and sheds new insights and targets for periodontitis therapeutics.

LncRNA UCA1 promotes pancreatic cancer cell migration by regulating mitochondrial dynamics via the MAPK pathway

PurposeLong non-coding RNA urothelial cancer associated 1 (UCA1) serves as an oncogene in various cancers. However, the mechanism underlying the role of UCA1 in pancreatic cancer remains unclear. This study aimed to explore the role of UCA1 in pancreatic cancer. MethodsThe expression and prognosis of UCA1 were analyzed using The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases. The results were validated by immunohistochemistry (IHC) and qRT-PCR. The biofunctions of UCA1 were analyzed using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA). The migration abilities and mitochondrial dynamics of PC cells were examined using the Transwell assay, mitochondrial membrane potential (MMP), and fluorescence. The mitochondrial-related protein and MAPK/ERK pathway markers were evaluated using western blotting. ResultsUCA1 expression was significantly higher in pancreatic cancer tissues than in normal tissues. High UCA1 expression indicated poor clinical outcomes and was associated with clinical features in patients with pancreatic cancer. Additionally, high UCA1 expression is a potential independent marker for poor prognosis. Subsequently, we demonstrated that UCA1 enhanced the migration capability, increased MMP, enhanced mitochondrial fusion, and inhibited mitochondrial autophagy in pancreatic cancer cells via the MAPK/ERK pathway. ConclusionUCA1 promotes the migration by regulating the mitochondrial dynamics of pancreatic cancer cells via the MAPK/ERK pathway. Our findings suggest that UCA1 may serve as a potential biomarker in pancreatic cancer prognosis.

Abstract LB039: Ovarian cancer cell-derived exosomal UCA1 reprograms glucose metabolism in stromal fibroblasts

Abstract Tumor progression is stringently regulated by the co-ordinated signaling between the tumor cells and the tumor microenvironment (TME) through diverse autocrine, paracrine and exocrine signaling mechanisms. Recent studies have shown that the exosomes that play a crucial role in the transfer of oncogenic macromolecules and oncometabolites to different cellular components of the TME can be targeted for therapy. Therefore, with the focus on identifying the exosomal long non-coding RNA (lncRNA) that can be therapeutically targeted, we investigated the profile of ovarian cancer cell derived lncRNAs and their functional role in ovarian cancer pathophysiology. Analysis of exosomes derived from a panel of high grade serous ovarian cancer cell lines indicated that UCA1 (Urothelial Cancer Associated 1) is the most abundantly packaged lncRNA in the exosomes. Similarly, exosomes from patient-derived ovarian cancer cells exhibited higher levels of UCA1. Ascites from ovarian cancer patients also indicated high levels of UCA1 in the ascites-derived exosomes. Results using PKH67-labelled ovarian cancer cell-derived exosomes and MRC5 fibroblast cell line indicated the exosomal transfer of UCA1 to the fibroblasts. Since paracrine signaling has been shown to induce metabolic reprogramming of peritumoral fibroblasts, we interrogated whether exosomal transfer of UCA1 could reprogram the glucose metabolism in the fibroblasts. Results from the Agilent Seahorse glycolytic stress assay indicate that the exosomal UCA1 promotes reprogramming of glucose metabolism in fibroblast cells while depleting of UCA1 in the exosomes failed to induce such metabolic reprogramming. Results from the analysis of the key glycolytic enzymes in the stromal fibroblasts, post-exosomal UCA1 uptake also corroborate this conclusion. Thus our results provide primary evidence that the exosomal-UCA1 induces pro-tumorigenic metabolic reprogramming in peri-tumoral fibroblasts. Together with the known oncogenic role of UCA1 in many different cancer cells, our findings indicate that targeting UCA1 could form a productive precision cancer strategy in ovarian cancer, targeting both the tumor cells and tumor stromal cells. This research was supported by the Department of Defense Ovarian Cancer Research Program Award W81XWH-18-1-0066, W81XWH-22-1-0415, the National Institute of General Medical Sciences grant P20 GM103639 and The National Cancer Institute of the National Institutes of Health grant P30 CA225520. Citation Format: Revathy Nadhan, Ji Hee Ha, Muralidharan Jayaraman, Srishti Kashyap, Danny N. Dhanasekaran. Ovarian cancer cell-derived exosomal UCA1 reprograms glucose metabolism in stromal fibroblasts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB039.

The role of long non-coding RNA UCA1 and MALAT1 in bladder cancer patients

BackgroundBladder cancer (BC) is considered the most prevalent tumor of the urinary tract, and incidence rates are rising globally. Long noncoding RNAs (lncRNAs) can serve as biomarkers to help the diagnosis and anticipation of recurrence in cancer patients as they are implicated in carcinogenesis and tumor progression. This study targeted to investigate the expression of lncRNA as urothelial cancer associated 1 (UCA1) as well as metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in the diagnosis of BC. Subjects and methodsThe study included 70 BC patients, 70 benign urinary tract diseases and 70 controls. The UCA1 and MALAT1 expressions were studied utilizing quantitative real-time polymerase chain reaction. ResultsThe UCA1 and MALAT1 expression were significantly increased in patients with BC than patients with chronic cystitis and controls (p < 0.001). The sensitivity and specificity for the diagnosis of BC for UCA1(87.14%, 97.14% respectively) and MALAT1 (92.86%, 90% respectively). Multivariate logistic regression analysis showed that UCA1 and MALAT1 were risk factors for the development of BC where the odds ratio (OR) for UCA1 was 1.262 (CI 1.004–1.587) and MALAT1 was 2.201(CI 1.036–4.676). ConclusionUCA1 and MALAT1 could be potential diagnostic biomarkers with good specificity and sensitivity in bladder cancer.

Over-expression of Long Non-coding RNA Urothelial Cancer-associated 1 as a Predictive Marker for Prostate Cancer.

To determine the expression of long non-coding RNA urothelial cancer-associated 1 (UCA1) by performing array-based quantitative polymerase chain reaction (PCR) and to identify the clinicopathological significance of UCA1 expression in prostate cancer using in situ hybridization (ISH) of surgically resected specimens. Array-based quantitative PCR was performed using 10 pairs of fresh malignant (prostate cancer) and normal tissue samples to determine UCA1 expression. Single-color RNA ISH of surgically resected prostate cancer specimens was performed using 70 formalin-fixed, paraffin-embedded tissue specimens to examine the clinicopathological significance of UCA1. Prostate cancer tissues exhibited higher levels of UCA1 expression than paired benign tissues. Furthermore, a correlation between high UCA1 expression and unfavourable clinicopathological characteristics, including advanced pathologic T stage, extraprostatic extension, presence of Gleason pattern 5, and involvement of the resection margins was observed. Notably, increased UCA1 expression significantly correlated with high- or very-high-risk patients, as defined by the 2023 National Comprehensive Cancer Network guidelines. UCA1 could be used as a novel diagnostic and prognostic biomarker for establishing an effective treatment protocol for prostate cancer.

Trophoblast Exosomal UCA1 Induces Endothelial Injury through the PFN1-RhoA/ROCK Pathway in Preeclampsia: A Human-Specific Adaptive Pathogenic Mechanism

Preeclampsia is regarded as an evolution-related disease that has only been observed in humans and our closest relatives, and the important factor contributing to its pathogenesis is endothelial dysregulation secondary to a stressed placenta. Hypoxia-inducible factor 1 subunit alpha (HIF1α), a highly conserved molecule in virtually all mammals, is regarded as a crucial regulator of the hypoxia adaptation and evolution. Persistent high expression of HIF1α in the placenta is one of the pathogenic mechanisms of preeclampsia. Therefore, human-specific molecules should link increased HIF1α to preeclampsia. We reported that urothelial cancer associated 1 (UCA1) is a potential mediator because it is a human-specific long noncoding RNA (lncRNA) that is upregulated in placental tissues and maternal serum from women with preeclampsia and is regulated by HIF1α. The cellular HIF1α-UCA1 pathway promoted the adaptation of trophoblasts to hypoxia by inducing vascular endothelial growth factor (VEGF) secretion and changes in the levels of key enzymes in glycolysis. On the other hand, circulating exosomal UCA1 secreted from stressed trophoblasts induced vascular endothelial dysfunction, especially excess ROS production, as measured by exosome extraction and a coculture system. At the molecular level, UCA1 physically bound to ubiquitin-specific peptidase 14 (USP14), which is a deubiquitinating enzyme, and UCA1 functioned as a scaffold to recruit USP14 to profilin 1 (PFN1), an actin-binding protein contributing to endothelial abnormalities and vascular diseases. This ternary complex inhibited the ubiquitination-dependent degradation of PFN1 and prolonged its half-life, further activating the RhoA/Rho-kinase (ROCK) pathway to induce ROS production in endothelial cells. Taken together, these observations suggest a role for the evolution-related UCA1 in the HIF1α-induced adaptive pathogenic mechanism of preeclampsia, promoting the survival of hypoxic trophoblasts and injuring maternal endothelial cells.

LncRNA UCA1/miR-182-5p/MGMT axis modulates glioma cell sensitivity to temozolomide through MGMT-related DNA damage pathways

Glioblastoma (GBM) is the most malignant subtype of gliomas. GBM resistance to temozolomide (TMZ) remains a huge challenge. O6-methylguanine-DNA methyltransferase (MGMT) is mainly responsible for repairing DNA alkylation damage caused by alkylating drugs such as TMZ; therefore, it has been regarded as the major cause of the resistance to TMZ. Hematoxylin and eosin (H&E) and immunohistochemical (IHC) staining were performed in tissue sections. LncRNA urothelial cancer-associated 1 (UCA1) knockdown was conducted via the transfection of the plasmid containing small interfering RNA (siRNA) targeting lncRNA UCA1. Cell viability and apoptosis were examined using MTT assay and flow cytometry. Nude mouse tumorigenicity assay was performed to detect tumor formation in vivo. MGMT expression and lncRNA UCA1 expression were increased in high-grade glioma tissues and cells. UCA1 knockdown in glioma cells enhanced TMZ efficacies in affecting glioma cell viability, cell apoptosis, MGMT protein level, and DNA damage markers invitro, as well as tumorigenesis invivo. Moreover, miR-182-5p targeted UCA1 and MGMT; miR-182-5p inhibited MGMT expression. Similar to UCA1 knockdown, miR-182-5p overexpression also promoted TMZ effects on glioma cell phenotype, MGMT expression level, and the levels of DNA damage markers. Under TMZ treatment, the efficacies of UCA1 knockdown in MGMT expression level and glioma cell sensitivity to TMZ were notably reversed after miR-182-5p overexpression. Taken together, we demonstrate the lncRNA UCA1/miR-182-5p/MGMT axis modulates glioma cell sensitivity to TMZ via MGMT-related DNA damage pathways.

Comment on 'Long noncoding RNA UCA1 promotes glutamine-driven anaplerosis of bladder cancer by interacting with hnRNP I/L to upregulate GPT2 expression' by Chen et al.'”

Bladder cancer is prevalent cancer worldwide with poor outcomes for patients with high-grade disease. Emerging evidence shows that alteration of metabolic status drives tumorigenesis in bladder cancer. As long noncoding RNA urothelial cancer associated 1 (UCA1) is known to play an essential role in cancer metabolisms, such as glycolysis and glutaminolysis. Chen et al. report the novel function of UCA1 in glutamine metabolism through interacting with heterogeneous nuclear ribonucleoproteins (hnRNPs) I and L (hnRNP I/L). This study reveals that UCA1 promotes glutamic pyruvate transaminase 2 (GPT2) expression at the transcription level in mechanistic studies. Inhibition of either UCA1, hnRNPI/L, or GPT2 significantly reduces bladder cancer tumor growth in the mice model. This work explores a new mechanism for glutamine metabolism and the novel therapeutic target of the UCA1-hnRNPI/L-GPT2 axis across malignancies.

Long noncoding RNA UCA1 promotes glutamine-driven anaplerosis of bladder cancer by interacting with hnRNP I/L to upregulate GPT2 expression

Long noncoding RNA urothelial cancer associated 1 (UCA1), initially identified in bladder cancer, is associated with multiple cellular processes, including metabolic reprogramming. However, its characteristics in the anaplerosis context of bladder cancer (BLCA) remain elusive. We identified UCA1 as a binding partner of heterogeneous nuclear ribonucleoproteins (hnRNPs) I and L, RNA-binding proteins (RBPs) with no previously known role in metabolic reprogramming. UCA1 and hnRNP I/L profoundly affected glycolysis, TCA cycle, glutaminolysis, and proliferation of BLCA. Importantly, UCA1 specifically bound to and facilitated the combination of hnRNP I/L to the promoter of glutamic pyruvate transaminase 2 (GPT2), an enzyme transferring glutamate to α-ketoglutarate, resulting in upregulated expression of GPT2 and enhanced glutamine-derived carbons in the TCA cycle. We also systematically confirmed the influence of UCA1 and hnRNP I/L on metabolism and proliferation via glutamine-driven anaplerosis in BLCA. Our study revealed the critical role of UCA1-mediated mechanisms involved in glutamine-driven anaplerosis and provided novel evidence that lncRNA regulates metabolic reprogramming in tumor cells.

The DNMT1-associated lncRNA UCA1 was upregulated in TK6 cells transformed by long-term exposure to hydroquinone and benzene-exposed workers via DNA hypomethylation.

Exposure to benzene or its metabolite hydroquinone (HQ) is a risk factor for a series of myeloid malignancies, and long noncoding RNAs play an important role in the process of pathogenesis. Urothelial cancer-associated 1 (UCA1) functions as an oncogene in the development of acute myeloid leukemia. However, the association between DNMT1 and UCA1 with benzene or HQ exposure has not been explored. We characterized UCA1 expression in cells briefly exposed to HQ (HQ-ST cells) and HQ-induced malignantly transformed (TK6-HT cells) treated with 5-aza-2'-deoxycytidine (5-AzaC) or trichostatin A (TSA). Compared to that in control cells, UCA1 expression was increased, whereas DNMT1 was decreased in HQ-ST cells and TK6-HT cells treated with 5-AzaC or TSA. Moreover, UCA1 expression was also upregulated and positively correlated with benzene exposure time in benzene-exposed workers. Furthermore, the expression of UCA1 was negatively associated with the DNA methylation level of its promoter in benzene-exposed workers. DNMT1 rather than DNMT3b knockout in TK6-HT cells activated the expression of UCA1 by inducing its promoter hypomethylation. These results suggest that benzene or HQ exposure leads to UCA1 upregulation via DNA hypomethylation in the UCA1 promoter, which is mediated by DNMT1.

Long non-coding RNA UCA1 enhances cervical cancer cell proliferation and invasion by regulating microRNA-299-3p expression.

The long non-coding RNA, urothelial cancer-associated 1 (UCA1) is an important regulator in several tumors. However, to the best of our knowledge, the clinical roles of UCA1 in cervical cancer remain unclear. Thus, the present study aimed to investigate the function and mechanism of UCA1 in cervical cancer. Reverse transcription-quantitative PCR analysis was performed to detect UCA1 and microRNA (miR)-299-3p expression in cervical cancer tissues and cell lines. The Cell Counting Kit-8 and Transwell assays were performed to assess cell proliferation and invasion, respectively. Furthermore, the dual-luciferase reporter assay was performed to confirm the association between UCA1 and miR-299-3p. Rescue experiments were performed to determine the mechanism of the UCA1/miR-299-3p axis. The results demonstrated that UCA1 expression was upregulated in cervical cancer tissues and cell lines. Furthermore, overexpression of UCA1 enhanced the proliferation and invasion of cervical cancer cells, the effects of which were reversed following UCA1 knockdown. Notably, UCA1 interacted with miR-299-3p and negatively regulated miR-299-3p expression. In addition, miR-299-3p expression was downregulated in cervical cancer tissues and cell lines. Overexpression of miR-299-3p suppressed the proliferation and invasion of cervical cancer cells, reversing the effects of UCA1 knockdown on cervical cancer cell proliferation. Taken together, the results of the present study suggest that UCA1 promotes cell proliferation and invasion by regulating miR-299-3p expression in cervical cancer.

LncRNA UCA1 Antagonizes Arsenic-Induced Cell Cycle Arrest through Destabilizing EZH2 and Facilitating NFATc2 Expression

Arsenic (As) is a widespread metalloid contaminant, and its internal exposure is demonstrated to cause serious detrimental health problems. Albeit considerable studies are performed to interrogate the molecular mechanisms responsible for As-induced toxicities, the exact mechanisms are not fully understood yet, especially at the epigenetic regulation level. In the present study, it is identified that long non-coding RNA (lncRNA) urothelial cancer associated 1 (UCA1) alleviates As-induced G2/M phase arrest in human liver cells. Intensive mechanistic investigations illustrate that UCA1 interacts with enhancer of zeste homolog 2 (EZH2) and accelerates the latter's protein turnover rate under normal and As-exposure conditions. The phosphorylation of EZH2 at the Thr-487 site by cyclin dependent kinase 1 (CDK1) is responsible for As-induced EZH2 protein degradation, and UCA1 enhances this process through increasing the interaction between CDK1 and EZH2. As a consequence, the cell cycle regulator nuclear factor of activated T cells 2 (NFATc2), a downstream target of EZH2, is upregulated to resist As-blocked cell cycle progress and cytotoxicity. In conclusion, the findings decipher a novel prosurvival signaling pathway underlying As toxicity from the perspective of epigenetic regulation: UCA1 facilitates the ubiquitination of EZH2 to upregulate NFATc2 and further antagonizes As-induced cell cycle arrest.

LncRNA UCA1 elevates the resistance of human leukemia cells to daunorubicin by the PI3K/AKT pathway via sponging miR-613.

Acute leukemia is a hematological malignant tumor. Long non-coding RNA urothelial cancer-associated 1 (UCA1) is involved in the chemo-resistance of diverse cancers, but it is unclear whether UCA1 is associated with the sensitivity of acute leukemia cells to daunorubicin (DNR). DNR (100 nM) was selected for functional analysis. The viability, cell cycle progression, apoptosis, and invasion of treated acute leukemia cells (HL-60 and U-937) were evaluated by cell counting kit-8 (CCK-8) assay, flow cytometry assay, or transwell assay. Protein levels were detected with Western blot analysis. Expression patterns of UCA1 and miR-613 were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). The relationship between UCA1 and microRNA-613 (miR-613) was verified by dual-luciferase reporter assay. We observed that UCA1 expression was elevated in HL-60 and U-937cells. DNR constrained viability, cell cycle progression, invasion, and facilitated apoptosis of HL-60 and U-937 cells in a dose-dependent manner, but these impacts mediated by DNR were reverted after UCA1 overexpression. MiR-613 was down-regulated in HL-60 and U-937 cells, and UCA1 was verified as a miR-613 sponge. MiR-613 inhibitor reversed DNR treatment-mediated effects on viability, cell cycle progression, apoptosis, and invasion of HL-60 and U-937 cells, but these impacts mediated by miR-613 inhibitor were counteracted after UCA1 inhibition. Notably, the inactivation of the PI3K/AKT pathway caused by DNR treatment was reversed after miR-613 inhibitor introduction, but this influence mediated by miR-613 inhibitor was offset after UCA1 knockdown. In conclusion, UCA1 up-regulation facilitated the resistance of acute leukemia cells to DNR via the PI3K/AKT pathway by sponging miR-613.

Exosomal lncRNA UCA1 modulates cervical cancer stem cell self-renewal and differentiation through microRNA-122-5p/SOX2 axis

BackgroundThere is growing evidence discussing the role of long non-coding RNAs (lncRNAs) in cervical cancer (CC). We performed this study to explore the impact of exosomal lncRNA urothelial cancer-associated 1 (UCA1) in CC stem cells by sponging microRNA-122-5p (miR-122-5p) and regulating SOX2 expression.MethodsCC stem cells (CD133+CaSki) and exosomes were extracted and identified. The synthesized UCA1- and miR-122-5p-related sequences were transfected into CaSki cells, CaSki cells-derived exosomes were extracted and then co-cultured with CD133+CaSki cells. The functional roles of UCA1 and miR-122-5p in self-renewal and differentiation ability of CC stem cells were determined using ectopic expression, knockdown/depletion and reporter assay experiments. An in vivo experiment was performed to verify the in vitro results.ResultsUp-regulated UCA1 and SOX2 and down-regulated miR-122-5p were found in CaSki-Exo. Exosomes promoted invasion, migration, proliferation and restrained apoptosis of CD133+CaSki cells. Silencing UCA1 or up-regulating miR-122-5p degraded SOX2 expression, and reduced invasion, migration and proliferation of CD133+CaSki cells while advanced apoptosis and suppressed the tumor volume and weight in nude mice.ConclusionOur study provides evidence that CaSki-Exo can promote the self-renewal and differentiation ability of CC stem cells while silencing UCA1 or up-regulating miR-122-5p restrains self-renewal and differentiation of CC stem cells.

Urothelial Cancer Associated 1 (UCA1) and miR-193 Are Two Non-coding RNAs Involved in Trophoblast Fusion and Placental Diseases.

A bioinformatics screen for non-coding genes was performed from microarrays analyzing on the one hand trophoblast fusion in the BeWo cell model, and on the other hand, placental diseases (preeclampsia and Intra-Uterine Growth Restriction). Intersecting the deregulated genes allowed to identify two miRNA (mir193b and miR365a) and one long non-coding RNA (UCA1) that are pivotal for trophoblast fusion, and deregulated in placental diseases. We show that miR-193b is a hub for the down-regulation of 135 cell targets mainly involved in cell cycle progression and energy usage/nutrient transport. UCA1 was explored by siRNA knock-down in the BeWo cell model. We show that its down-regulation is associated with the deregulation of important trophoblast physiology genes, involved in differentiation, proliferation, oxidative stress, vacuolization, membrane repair and endocrine production. Overall, UCA1 knockdown leads to an incomplete gene expression profile modification of trophoblast cells when they are induced to fuse into syncytiotrophoblast. Then we performed the same type of analysis in cells overexpressing one of the two major isoforms of the STOX1 transcription factor, STOX1A and STOX1B (associated previously to impaired trophoblast fusion). We could show that when STOX1B is abundant, the effects of UCA1 down-regulation on forskolin response are alleviated.

LncRNA UCA1 induced by SP1 and SP3 forms a positive feedback loop to facilitate malignant phenotypes of colorectal cancer via targeting miR-495

AimsLong noncoding RNA (LncRNA) urothelial cancer associated 1 (UCA1) was dysregulated in colorectal cancers (CRC) and promoted tumor progression of CRC. The aims of this study are to further investigate the underlying mechanism. Main methodsShort hairpin RNAs (shRNAs) were applied for gene knockdown. microRNA mimic and pcDNA-UCA1 plasmids were transfected for miR-495 and UCA1 overexpression, respectively. MTT was applied to determine cell viability and sensitivity of 5-fluorouracil (FU). Transwell assays were performed to evaluate cell migration/invasion. Angiogenesis was evaluated by tube formation. Western blotting and quantitative PCR were utilized for protein and mRNA detection, respectively. The interaction of UCA1, miR-495 and SP1/SP3 were explored by dual-luciferase assay. RNA pulldown was adopted to determine the UCA1/miR-495 interaction. Key findingsUCA1 was significantly upregulated in CRC tissues. UCA1 enhanced cell proliferation, migration/invasion, angiogenesis, epithelial-mesenchymal transition, and resistance to 5-FU in CRC cell lines. MiR-495 was inversely correlated to the expression of UCA1. The results indicated that UCA1 sponged miR-495, leading to the disinhibition of SP1/SP3 expression. SP1/SP3 induced the expression of DNA methyltransferases and, in turn, contributed to UCA1 mediated tumor-promoting actions. Reduction of SP1/SP3 exerted anti-cancer effects, which can be reversed by forced expression of UCA1. SignificanceUCA1-miR-495-SP1/SP3 axis is dysregulated in CRC and contributed to malignant phenotypes of CRC. UCA1-SP1/SP3 may form a positive feedback loop in CRC.

The common region of lncRNAs UCA1 and UCA1α contributes to the bladder cancer tumorigenesis.

Accumulating evidence suggests that the long noncoding RNAs (lncRNAs) urothelial cancer associated 1 (UCA1) and urothelial cancer associated 1α (UCA1α) play a critical role in the development and progression of bladder cancer. However, the detailed mechanism, in particular the role of the common region of UCA1 and UCA1α, termed UCA1 same sequence (USS, approximately 1265 bp), remains unknown. To address this, the full length of 1265 bp USS was obtained initially, and then produced pcDNA3.1(+)-based overexpression vectors and designed siRNAs for UCA1, UCA1α, and USS, and assessed their effects on the human bladder cancer cell lines UMUC3 (which does not express UCA1 and UCA1α) and 5637 (which has high expression of UCA1 and UCA1α), respectively. Overexpression of UCA1, UCA1α, and USS significantly increased bladder cancer cell proliferation and the number of S-phase cells, but significantly decreased cell apoptosis and the number of G0/G1 phase cells. Consistently, silencing UCA1 and USS significantly decreased bladder cancer cell proliferation and the number of S phase cells, but significantly increased apoptosis and the number of G0/G1 phase cells. These results suggest that USS, the common region of UCA1 and UCA1α, can promote bladder cancer tumorigenesis by increasing cell proliferation and decreasing cell apoptosis in a similar way to UCA1 and UCA1α lncRNAs. This not only transforms our understanding of the role of UCA1, UCA1α, and their common region in tumorigenesis, but may also provide a new target for future clinical applications.

Identification of Urothelial Cancer-Associated 1 (UCA1) as a diagnostic biomarker of pre-eclampsia via regulating microRNA-16 and its downstream signaling pathway

IntroductionIn this study, we studied the clinical value of using urothelial cancer-associated 1 (UCA1) and microRNA-16 (miR-16) as biomarkers for the diagnosis of pre-eclampsia (PE). Also, we compared the diagnostic values of miR-16, UCA1 and pregnancy-associated plasma protein-A (PAPP-A) in PE. Furthermore, we investigated the interaction between miR-16 and UCA1/PAPP-A.Material and methods128 PE patients and 172 healthy pregnant women were enrolled in this study. Receiver operating characteristic (ROC) analysis was carried out to predict the diagnostic values of UCA1, miR-16 and PAPP-A in PE. Enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (PCR), Western blot analysis, immunohistochemistry (IHC) assay, computational analysis, and luciferase assay were conducted to measure the differential expression of UCA1, miR-16, and PAPP-A while establishing a signaling pathway of UCA1/miR-16/PAPP-A.ResultsCompared with miR-16 and PAPP-A, UCA1 exhibited a better value in the diagnosis of PE. The expression of PAPP-A and UCA1 was down-regulated while the expression of miR-16 was up-regulated in patients with PE, especially in patients with HELLP pregnancies. Moreover, UCA1 was identified as a sponge of miR-16, while PAPP-A mRNA was identified as a virtual target gene of miR-16. Finally, a negative regulatory relationship was observed between the expression of miR-16 and UCA1 or PAPP-A, while the expression of UCA1 and PAPP-A were positively related.ConclusionsTaken together, UCA1 could be used as a more valuable biomarker in the diagnosis of PE. Meanwhile, the reduced expression of UCA1 could exert a positive effect by reducing the expression of PAPP-A in the pathogenesis of PE.