lncRNA Colorectal Neoplasia Differentially Expressed Research Articles

Long term exercise-derived exosomal LncRNA CRNDE mitigates myocardial infarction injury through miR-489-3p/Nrf2 signaling axis

Myocardial infarction (MI) is a cardiovascular disease and troubles patients all over the world. Exosomes produced after long-term exercise training were discovered to mediate intercellular communication and alleviate MI-induced heart injury. However, the detailed roles of long-term exercise-derived exosomal long noncoding RNAs (LncRNAs) in MI remain uncovered. In this study, we collected and identified long-term exercise-derived exosomes, and established MI or hypoxia/reoxygenation (H/R) model after LncRNA colorectal neoplasia differentially expressed (CRNDE) depletion. This work proved that LncRNA CRNDE was highly expressed in long-term exercise-derived exosomes (p = 0.0078). CRNDE knockdown increased cardiomyocytes apoptosis and oxidative stress (p = 0.0036), and suppressed MI progress (p = 0.0005). CRNDE served as the sponge of miR-489-3p to affect Nrf2 expression (p = 0.0001). MiR-489-3p inhibition effectively reversed the effects of CRNDE depletion on hypoxia cardiomyocytes (p = 0.0002). These findings offered a promising therapeutic option for the treatment of MI.

The long non-coding RNA CRNDE promotes osteosarcoma proliferation and migration by sponging miR-136-5p/MRP9 axis.

BackgroundThe long-noncoding RNA colorectal neoplasia differentially expressed (CRNDE) gene has been found to be upregulated in several solid tumors. Whether CRNDE affects osteosarcoma (OS) and its underling mechanism remains unknown.MethodsTumor tissues and corresponding normal tissues were collected from 45 patients with OS. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was applied to determine lncRNA CRNDE level in the tissues. Participants were divided into a high CRNDE group and a low CRNDE group according to the median value of lncRNA CRNDE expression detected by in situ hybridization (ISH). The differences between high and low expression of lncRNA CRNDE in patients were compared clinically by chi-square test. Kaplan-Meier survival analysis was applied to analyze the relationship between lncRNA CRNDE expression and patient survival. Subsequently, silencing or overexpression of lncRNA CRNDE were performed in MG63 and 143B cell lines, qRT-PCR was applied to verify the expression of lncRNA CRNDE, miR-136-5p, and MRP9; dual-luciferase reporter assay was used to evaluate the targeting relationship between miR-136-5p, lncRNA CRNDE, and Cell Counting Kit-8 (CCK8), wound-healing, and Transwell assays were used to analyze for cell proliferation, migration, and invasion, respectively, and western blot was used to detect expression in cells.ResultsThe expression of CRNDE in OS tissues was higher than that in normal tissues. High lncRNA CRNDE expression was significantly associated with clinical stage, lung metastasis, and poor prognosis in OS patients. Additionally, overexpression of lncRNA CRNDE promoted proliferation and migration of OS cells. Bioinformatics analysis showed that lncRNA CRNDE competitively inhibited miR-136-5p through acting as a competitive endogenous RNA (ceRNA). It was also revealed that miR-136-5p is a binding target gene of lncRNA CRNDE and that MRP9 is involved in this process as a downstream target gene of miR-136-5p.ConclusionsThe lncRNA CRNDE promotes the proliferation and migration of OS cells by regulating the miR-136-5p/MRP9 pathway, and lncRNA CRNDE can be a significant marker of OS prognosis.

LncRNA CRNDE hinders the progression of osteoarthritis by epigenetic regulation of DACT1.

Osteoarthritis (OA) is mainly characterized by articular cartilage degeneration, synovial fibrosis, and inflammation. LncRNA CRNDE (colorectal neoplasia differentially expressed) has been reported to be down-regulated in age-related OA, but its role in injury-induced OA needs to be further explored. In this study, an OA rat model was established using anterior cruciate ligament transection, and the adenovirus-mediated CRNDE overexpression (Ad-CRNDE) or DACT1 (dapper antagonist of catenin-1) interference (sh-DACT1) vectors were administered by intraarticular injection. Moreover, chondrocyte‑like ATDC5 cells were treated with IL-1β (10ng/mL) to simulate OA conditions in vitro. We found that overexpression of CRNDE alleviated cartilage damage and synovitis in OA rats, and suppressed IL-1β-induced apoptosis, inflammation, and extracellular matrix (ECM) degradation in chondrocyte‑like ATDC5 cells, while silencing DACT1 effectively antagonized the protective effect of CRNDE both in vivo and in vitro. Mechanism studies revealed that DACT1 could act as a downstream target of CRNDE. By recruiting p300, CRNDE promoted the enrichment of H3K27ac in the DACT1 promoter, thus promoting DACT1 transcription. In addition, CRNDE hindered the activation of the Wnt/β-catenin pathway in IL-1β-stimulated cells by inducing DACT1 expression. In conclusion, CRNDE promoted DACT1 expression through epigenetic modification and restrained the activation of Wnt/β-catenin signaling to impede the progression of OA.

The lncRNA CRNDE is regulated by E2F6 and sensitizes gastric cancer cells to chemotherapy by inhibiting autophagy.

Chemotherapy is an important treatment for gastric cancer (GC), but the primary and secondary drug resistance of tumours to chemotherapy seriously affects its curative effect. In recent years, the relationship between long noncoding RNAs (lncRNAs) and malignant tumours has received increasing attention. Based on accumulating evidence, lncRNAs are involved in the chemoresistance of GC, but the underlying mechanisms remain unclear. In this study, we identified the lncRNA colorectal neoplasia differentially expressed (CRNDE) as an important regulator of autophagy-associated chemoresistance in GC. Mechanistically, overexpression of CRNDE inhibits autophagy and induces apoptosis, thereby sensitizing GC cells to chemotherapy drugs. Moreover, E2F6, a classical transcriptional inhibitor, is confirmed to be upregulated in GC and represses the expression of CRNDE. The E2F6-CRNDE axis is clinically related to chemoresistant GC and poor outcomes in patients with advanced GC. Our findings suggest that the E2F6-CRNDE axis is a viable therapeutic target to protect against chemoresistance in GC.

LncRNA CRNDE Exacerbates IgA Nephropathy Progression by Promoting NLRP3 Inflammasome Activation in Macrophages

ABSTRACT Background Activation of NLRP3 inflammasome in macrophages contributes greatly to IgA nephropathy (IgAN) progression. This study intended to investigate the underlying mechanism of NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome activation in the development of IgAN. Methods We examined the expression levels of colorectal neoplasia differentially expressed (CRNDE), NLRP3 inflammasome-related proteins in peripheral blood mononuclear cells (PBMCs) and J774A.1 cells and detected inflammatory cytokine levels in the serum of IgAN patients and cell supernatants of in vitro IgAN model. RNA pull-down and RNA immunoprecipitation (RIP) experiments were conducted to evaluate the interaction between CRNDE and NLRP3. Then, the ubiquitin level of NLRP3 and its binding ability to TRIM family member 31 (TRIM31) were determined. Results Compared with the control group, the expressions of CRNDE and NLRP3 inflammasome-related proteins in PBMCs and J774A.1 cells and levels of IL-1β, TNF-α and IL-12 in serum of IgAN patients and cell supernatants of IgA-IC-induced J774A.1 cells were all increased. CRNDE silencing down-regulated NLRP3 inflammasome-related proteins and the levels of IL-1β, TNF-α and IL-12 in cell supernatants, while NLRP3 overexpression reversed these effects. Additionally, CRNDE could interact with NLRP3 and promote NLRP3 expression. Furthermore, inhibition of CRNDE reduced NLRP3 protein level and promoted TRIM31-mediated NLRP3 ubiquitination and degradation. Conclusion CRNDE exacerbates IgA nephropathy progression through restraining ubiquitination and degradation of NLRP3 and facilitating NLRP3 inflammasome activation in macrophages.

Exosomal transfer of activated neutrophil-derived lncRNA CRNDE promotes proliferation and migration of airway smooth muscle cells in asthma.

Activated neutrophil-derived exosomes reportedly contribute to the proliferation of airway smooth muscle cells (ASMCs), thereby aggravating the airway wall remodeling during asthma; however, the specific mechanism remains unclear. Lipopolysaccharide (LPS)-EXO and si-CRNDE-EXO were extracted from the media of human neutrophils treated with LPS and LPS + si-CRNDE (a siRNA targets long non-coding RNA CRNDE), respectively. Human ASMCs were co-cultured with LPS-EXO or si-CRNDE-EXO, and cell viability, proliferation and migration were measured. The interplay of colorectal neoplasia differentially expressed (CRNDE), inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) and nuclear receptor subfamily 2 group C member 2 (TAK1) was explored using RNA immunoprecipitation (RIP) and Co-IP assays. A mouse model of asthma was induced using ovalbumin. CRNDE was upregulated in LPS-EXO and successfully transferred from LPS-treated neutrophils to ASMCs through exosome. Mechanically, CRNDE loaded in LPS-EXO reinforced TAK1-mediated IKKβ phosphorylation, thereby activating the nuclear factor kappa B (NF-κB) pathway. Functionally, silencing CRNDE in LPS-EXO, an IKKβ inhibitor, and an NF-κB inhibitor all removed the upregulation of cell viability, proliferation and migration induced by LPS-EXO in ASMCs. In the end, the in vivo experiment demonstrated that CRNDE knockdown in neutrophils effectively reduced the thickness of bronchial smooth muscle in a mouse model for asthma. Activated neutrophils-derived CRNDE was transferred to ASMCs through exosomes and activated the NF-κB pathway by enhancing IKKβ phosphorylation. The latter promoted the proliferation and migration of ASMCs and then contributed to airway remodeling in asthma.

Long non-coding RNA colorectal neoplasia differentially expressed correlates negatively with miR-33a and miR-495 and positively with inflammatory cytokines in asthmatic children.

ObjectivesIt is reported that long non‐coding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) targets microRNA (miR)‐33a, miR‐181a and miR‐495 to regulate inflammation process, while few studies report their clinical application for paediatric asthma management. Therefore, this study aimed to explore the interaction of lncRNA CRNDE with miR‐33a, miR‐181a and miR‐495, as well as their correlation with inflammation, exacerbation risk and severity in paediatric patients with asthma.MethodsAsthmatic exacerbation children (N = 65), asthmatic controlled children (N = 65) and controls (N = 65) were recruited. LncRNA CRNDE, miR‐33a, miR‐181a and miR‐495 expressions in peripheral blood mononuclear cells were detected by RT‐qPCR. Besides, serum inflammatory cytokines (including TNF‐α, IL‐1β, IL‐6 and IL‐17) were determined by enzyme‐linked immunosorbent assay (ELISA).ResultsLncRNA CRNDE, miR‐33a and miR‐495 expressions were different, while miR‐181a expression was similar among asthmatic exacerbation children, asthmatic controlled children and controls. Moreover, lncRNA CRNDE negatively correlated with miR‐33a and miR‐495 in asthmatic exacerbation children and asthmatic controlled children, but not in controls. Further analyses showed that lncRNA CRNDE positively correlated with TNF‐α, IL‐1β, IL‐17 and exacerbation severity, while it negatively correlated with FEV1/FVC in asthmatic exacerbation children. Meanwhile, miR‐33a, miR‐181a and miR‐495 all negatively correlated with some individual inflammatory cytokines, while only miR‐33a negatively correlated with exacerbation severity in asthmatic exacerbation children.ConclusionLncRNA CRNDE correlates negatively with miR‐33a and miR‐495 and positively with inflammatory cytokines in asthmatic children.

Implication of the Long Non-Coding RNA Crnde in Multiple Myeloma

Multiple myeloma (MM) is a malignancy of antibody-secreting plasma cells which remains incurable, despite significant improvements in treatment and patient care. MM is characterized by a wide clinical and prognostic spectrum, even within groups bearing the same primary initiating cytogenetic event, for which the molecular mechanisms responsible remain poorly understood. Long non-coding RNAs (lncRNAs; broadly defined as non-coding RNAs of > 200 nt) have recently emerged as an important class of regulatory molecules, exercising diverse functions in normal cells and tissues, and are increasingly implicated in tumorigenesis and cancer progression. However at present the contribution of lncRNAs to the progression and clinical variability of MM is largely unknown.In order to indentify lncRNAs potentially implicated in the etiology of MM, we have taken a data mining approach to compare the expression profile of lncRNAs between CD138+ plasmocytes from healthy individuals with those from MM patients. Patients were stratified according to the presence of the t(4;14) or t(11;14) translocations and a third group bearing neither of these cytogenetics events. This analysis revealed 110 lncRNAs that are differentially expressed in CD138+ plasma cells from newly diagnosed MM patients compared to control subjects. An interesting candidate to emerge from this analysis is Colorectal Neoplasia Differentially Expressed (CRNDE), a lncRNA that has previously been implicated in other hematological malignancies and diverse solid tumors, by promoting growth and inhibiting apoptosis potentially via a physical association with epigenetic modifying complexes (PRC2 and CoREST). Consistent with these observations in other malignancies, our findings suggest that CRNDE is also likely to have a role in the progression of MM. Expression of CRNDE increases with disease progression from the pre-malignant MGUS stage, through the stage of smoldering myeloma and reaches highest levels at progression to overt symptomatic myeloma. Further, survival analysis indicates that high expression of CRNDE is associated with poor overall survival, in particular in high risk t(4;14)+ myeloma patients.To directly test the role of CRNDE in MM and to investigate downstream oncogenic pathways, we are using CRISPR/Cas9 to delete the major nuclear and cytoplasmic isoforms of CRNDE in MM cell lines. Our first results indicate that the deletion of CRNDE exerts a dose-dependent repressive effect on myeloma cell growth, supporting the idea that CRNDE is an important determinant of disease progression in MM. CRISPER/Cas9 deletion is being carried out in both a t(4;14) and non t(4;14) context. We have further investigated the effect of CRNDE deletions on the sensitivity of MM cells to therapeutic agents and are studying the interaction of CRNDE with Polycomb group proteins.In summary, our results using CRISPR/Cas9-mediated deletion of the lncRNA CRNDE in MM cell lines together with the clinical data on patient survival strongly suggest that CRNDE plays an important role in MM disease progression and outcome. This work has the potential to translate into clinical benefit in a number of ways, notably by providing a useful biomarker of MM. Further, the development of molecular inhibitors of lncRNAs is a rapidly expanding field opening the way to the development of novel therapeutic strategies based on lncRNAs. DisclosuresNo relevant conflicts of interest to declare.

Long noncoding RNA CRNDE functions as a diagnostic and prognostic biomarker in osteosarcoma, as well aspromotes itsprogression via inhibition of miR-335-3p.

This study was performed to evaluate the diagnostic and prognostic value, as well as the role of long-chain noncoding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) in osteosarcoma (OS). A quantitative real-time polymerase chain reaction assay was to determine lncRNA CRNDE and microRNA-335-3p(miR-335-3p) expressions. The Kaplan-Meier analysis was to analyze the relationship between lncRNA CRNDE expression and survival in patients with OS. Receiver operating characteristic curves were to evaluate the diagnostic value of lncRNA CRNDE in OS. Bioinformatics analysis and luciferase reporter assays were used to predict and confirm the relationship between lncRNA CRNDE and miR-335-3p. Cell counting Kit-8 and transwell migration assays assessed the role of lncRNA CRNDE and miR-335-3p in OS cells. lncRNA CRNDE expression was upregulated and miR-355-3p expression was downregulated in OS. In patients with OS, low lncRNA CRNDE expression demonstrated higher overall survival, whereas high lncRNA CRNDE expression was an independent poor prognostic factor. Furthermore, increased lncRNA CRNDE expression was associated with distant metastasis and the tumor-node-metastasisstage in patients with OS, which can be considered as an independent diagnostic biomarker in OS. We revealed that miR-335-3p was the target of lncRNA CRNDE. It also demonstrated that knockdown of lncRNA CRNDE inhibited OS cell proliferation, migration, and invasion,and inhibition of miR-355-3p promoted this effect. Finally, miR-335-3p partially mediated the stimulatory effects of lncRNA CRNDE in OS. We demonstrated that lncRNA CRNDE is a potential diagnostic and prognostic biomarker for OS, and the lncRNA CRNDE/miR-335-3p axis participates in OS progression.

Protective effect of lncRNA CRNDE on myocardial cell apoptosis in heart failure by regulating HMGB1 cytoplasm translocation through PARP-1.

Long non-coding RNAs (lncRNAs) are bound up with the regulation of variousdiseases. Here, we probed into the effect of lncRNA colorectal neoplasia differentially expressed (CRNDE) on heart failure (HF). The pathological alterations and cell apoptosis of hearttissues were observed by hematoxylin-eosin and TUNEL staining. The viability or apoptosis of mouse myocardial cells HL-1 was tested by XTT or flow cytometry. The interaction between lncRNA CRNDE and poly-ADP-ribose polymerase 1 (PARP-1) was verified by RNA immunoprecipitation and RNA pull-down. The stability of the PARP-1 protein and the acetylation level of high mobility group box-1 (HMGB1) were determined by cycloheximide-chase and immunoprecipitation, respectively. LncRNA CRNDE expressionwas decreased in HF mice tissues and doxorubicin (Dox)-treated HL-1 cells, whereas PARP-1 and HMGB1 wereincreased. The overexpression of lncRNA CRNDE restrained HL-1 cell apoptosis induced by Dox. Moreover, the interaction between CRNDE and PARP-1 was corroborated, CRNDE negatively regulated PARP-1 expression, and the overexpression of CRNDE reducedPARP-1 protein stability. In HL-1 cells, PARP-1 positively regulated the acetylation level and cytoplasm translocation of HMGB1. CRNDE restrained Dox-induced apoptosis in mouse myocardial cells via the PARP-1/HMGB1 pathway.

LncRNA CRNDE is Upregulated in Glioblastoma Multiforme and Facilitates Cancer Progression Through Targeting miR-337-3p and ELMOD2 Axis.

IntroductionColorectal neoplasia differentially expressed (CRNDE) was reported to promote carcinogenesis in several cancers. However, the role of CRNDE in glioblastoma multiforme (GBM) needs to be further explored.MethodsCRNDE expression levels in GBM tissues and cells were explored using real-time quantitative PCR at first. Effects of CRNDE on GBM cell behaviors were detected by conducting in vitro experiments. Interactions of CRNDE, microRNA-337-3p (miR-337-3p), and ELMO domain containing 2 (ELMOD2) were verified by bioinformatics analysis tools and dual-luciferase reporter assay. Expression correlations of CRNDE and ELMOD2 in GBM tissues were analyzed at GEPIA website.ResultsCRNDE expression was upregulated in GBM tissues and cells compared with normal counterparts. CRDNE knockdown inhibits proliferation and migration, but promotes apoptosis in GBM cell, while CRNDE overexpression caused opposite effects. Mechanisms exploration indicated CRNDE serves as sponge of miR-337-3p to upregulate ELMOD2 expression. Furthermore, we showed CRNDE and ELMOD2 were positively correlated in GBM tissues.DiscussionIn conclusion, our study highlighted the importance of CRNDE/miR-337-3p/ELMOD2 axis in GBM progression and offered novel strategies for GBM treatment.

LncRNA CRNDE facilitates epigenetic suppression of CELF2 and LATS2 to promote proliferation, migration and chemoresistance in hepatocellular carcinoma

Our study aimed to investigate the expression, functional significance, and related mechanism of long noncoding RNA CRNDE (colorectal neoplasia differentially expressed) in hepatocellular carcinoma (HCC) pathogenesis. The resulted revealed that CRNDE was significantly overexpressed in HCC tissues and cell lines, and was statistically correlated with poor clinical outcome. CRNDE knockdown markedly decreased HCC cell proliferation, migration, and chemoresistance. In addition, in vivo experiments confirmed the suppressive effect of CRNDE knockdown on HCC progression. Mechanically, CRNDE directly bound to EZH2 (enhancer of zeste homolog), SUZ12 (suppressor of zeste 12), SUV39H1, and mediated their inhibition of tumor suppressor genes, including CUGBP Elav-like family member 2 (CELF2) and large tumor suppressor 2 (LATS2). CELF2 exerted tumor suppressive effect in HCC and was involved in CRNDE-mediated oncogenic effect. In addition, the oncogenic effects of CRNDE on HCC proliferation, migration and tumorigenesis, as well as its inhibition of Hippo pathway were abolished by LATS2 overexpression. Together, our work demonstrated the importance of CRNDE in HCC progression and elucidated the underlying molecular mechanisms. These findings provided new insights into HCC pathogenesis and chemoresistance mediated by CRNDE.

Knockdown of LncRNA CRNDE suppresses proliferation and P-glycoprotein-mediated multidrug resistance in acute myelocytic leukemia through the Wnt/β-catenin pathway.

Mechanisms involved in non-coding RNAs have been implicated in multidrug resistance (MDR) of acute myeloid leukemia (AML). Long non-coding RNA (lncRNAs) colorectal neoplasia differentially expressed (CRNDE) is reported to be involved in the malignant progression in AML. The purpose of the present study is to explore the roles and potential molecular mechanism of CRNDE in the MDR in AML. In our study, we confirmed that the expression of CRNDE was significantly up-regulated in patients with AML, especially in AML patients after adriamycin (ADR)-based chemotherapy. Spearman correlation analysis showed a positive correlation between the levels of CRNDE and MDR1 in AML patients after ADR-based chemotherapy. Moreover, CRNDE was up-regulated in AML cells, especially in ADR-resistant AML cells. Multidrug resistance protein 1 (MDR1)/p-glycoprotein (P-gp) levels were significantly increased in ADR-resistant AML cells, compared with parental AML cells. CRNDE down-regulation inhibited cell proliferation, promoted apoptosis, reduced Ki67 expression and enhanced cleaved caspase-3 expression in AML and ADR-resistant AML cells. In addition, CRNDE knockdown led to down-regulation of P-gp/MDR1, β-catenin, c-Myc and cyclinD1 expression, and enhanced the drug sensitivity to ADR in ADR-resistant AML cells. In conclusion, knockdown of CRNDE suppresses proliferation and P-gp-mediated MDR in ADR-resistant AML cells via inhibiting the Wnt/β-catenin pathway, suggesting that repression of CRNDE might be a therapeutic target to reverse MDR of ADR-resistant AML cells.

LncRNA CRNDE is activated by SP1 and promotes osteosarcoma proliferation, invasion, and epithelial-mesenchymal transition via Wnt/β-catenin signaling pathway.

Long noncoding RNAs (lncRNAs) were identified as a vital part in the development and progression of cancer in recent years. Colorectal neoplasia differentially expressed (CRNDE), a lncRNA, functions as an oncogene in some malignant neoplasias, but its role in the progression of osteosarcoma (OS) is still poorly understood. To dissect the difference in the expression of CRNDE, quantitative real-time polymerase chain reaction was utilized to evaluate it in OS tissues and cell lines (U2OS, MG63, and MNNG/HOS) compared with that in the adjacent normal tissues/osteoblast cells (hFOB1.19). The role of CRNDE in OS lines was assessed using Cell Counting Kit-8, colony formation, 5-ethynyl-2'-deoxyuridine staining, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling staining, flow cytometry, Transwell assays, and Western blot, respectively. The results demonstrated that the expression of CRNDE was high in OS tissues and cell lines, and partly induced by SP1. CRNDE knockdown attenuated OS cell proliferation and invasion and induced apoptosis and G0/G1 arrest. Moreover, the expression of mesenchymal markers N-cadherin, Vimentin and Snail were downregulated, while the expression of epithelial markers E-cadherin and ZO-1 were conversely upregulated due to CRNDE knockdown. The mechanistic investigations showed that CRNDE promoted glycogen synthase kinase-3β phosphorylation to activate the Wnt/β-catenin pathway. The results suggested that lncRNA CRNDE indeed contributed to OS proliferation, invasion, and epithelial-mesenchymal transition, working as an oncogene, demonstrating that lncRNA CRNDE may be a valid therapeutic target for the OS.

Effect of lncRNA CRNDE targeting miR-384 on radiosensitivity of colorectal cancer cells

Objective To study the effect of LncRNA CRNDE on radiosensitivity of colorectal cells and underlying mechanism. Methods Colorectal cancer HT-29 cells were transfected with CRNDE shRNA and the interference efficiency was determined by Real time PCR. HT-29 cells transfected with CRNDE shRNA or co-transfected with CRNDE shRNA and miR-384 inhibitor were irradiated at 8 Gy dose, then cell proliferation and apoptosis were detected by MTT and flow cytometry assay, respectively, and cell radiosensitivity was evaluated by cloning assay. It was predicted by a bioinformatics software that CRNDE and miR-384 have complementary binding sites, and this was identified by a luciferase reporting system. Results CRNDE shRNA reduced the expression of CRNDE in HT-29 cells(1.00±0.08 vs. 0.42±0.06, t=10.051, P<0.05). Both CRNDE shRNA and radiation inhibited the proliferation and induced apoptosis of HT-29 cells, and their combination treatment had synergistic effect in apoptosis induction [Apoptosis rates: (2.27±0.13)%, (23.58±2.35)%, (26.91±2.81)%, (36.84±3.24)%, F=24.660, P<0.05; A values: 0.45±0.06, 0.30±0.02, 0.28±0.03, 0.20±0.02, F=106.207, P<0.05]. Transfection of CRNDE shRNA increased the radiosensitivity of HT-29 cells with a radiosensitization ratio of 1.374. CRNDE negatively regulated the expression of its target miR-384. The miR-384 inhibitor antagonized the effect of CRNDE shRNA on proliferation inhibition and apoptosis promotion of radiation-treated colorectal cancer cells. Conclusions Down-regulation of LncRNA expression enhances the radiosensitivity of colorectal cancer cells by regulating miR-384 expression. Key words: Colorectal cancer; Colorectal neoplasia differentially expressed(CRNDE); miR-384; Radiosensitivity

Down-regulation of LncRNA CRNDE aggravates kidney injury via increasing MiR-181a-5p in sepsis

Long non-coding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) is reported to be linked to inflammation and cell apoptosis. However its role in sepsis induced kidney injury remains unclear. This study aims to explore the possible mechanism of CRNDE in kidney injury induced by sepsis. In vivo urine-derived sepsis (US) rat model and in vitro LPS-induced HK-2 and HEK293 cells were established. Kidney function was measured in rats from different groups. Relative levels of tumor necrosis factor-α (TNF-α) and interleukin-1β(IL-1β) in kidney tissue were detected via Enzyme-linked immune sorbent assay (ELISA). Then we up- or down-regulated CRNDE and miRNA-181a-5p expression in the cells. The biological influence of CRNDE and miR-181a-5p on cells was studied using CCK-8 assay and Annexin V assay. Interaction between CRNDE and miR-181a-5p was determined by bioinformatics analysis, RT-PCR, and dual luciferase reporter assay. Peroxisome proliferator-activated receptor-α (PPARα) and cell apoptosis related molecules were detected by western blot. We demonstrated that CRNDE was markedly down-regulated while miR-181a-5p was significantly up-regulated in sepsis models. CRNDE interacted with miR-181a-5p, and negatively regulated its expression level. CRNDE knockdown in rats increased the urea nitrogen and serum creatinine in plasma. Knockdown of CRNDE or transfection of miR-181a-5p significantly inhibited proliferation and promoted apoptosis of HK-2 and HEK293 cells, while overexpression of CRNDE and transfection of miR-181a-5p inhibitors had opposite effects. For mechanism, miR-181a-5p directly targeted the 3′ untranslated region of PPARα, and depressed its protein level, and PPARα was regulated indirectly by CRNDE. We concluded that CRNDE protected renal cell from sepsis-induced injury via miR-181a-5p/PPARα pathway.

Transcriptome analysis-identified long noncoding RNA CRNDE in maintaining endothelial cell proliferation, migration, and tube formation

Obesity is a leading risk factor for type-2 diabetes. Diabetes often leads to the dysregulation of angiogenesis, although the mechanism is not fully understood. Previously, long noncoding RNAs (lncRNAs) have been found to modulate angiogenesis. In this study, we asked how the expression levels of lncRNAs change in endothelial cells in response to excessive palmitic acid treatment, an obesity-like condition. Bioinformatics analysis revealed that 305 protein-coding transcripts were upregulated and 70 were downregulated, while 64 lncRNAs were upregulated and 46 were downregulated. Gene ontology and pathway analysis identified endoplasmic reticulum stress, HIF-1 signaling, and Toll-like receptor signaling as enriched after palmitic acid treatment. Moreover, we newly report enrichment of AGE-RAGE signaling pathway in diabetic complications, IL-17 signaling, and cysteine and methionine metabolism by palmitic acid. One lncRNA, Colorectal Neoplasia Differentially Expressed (CRNDE), was selected for further investigation. Palmitic acid induces CRNDE expression by 1.9-fold. We observed that CRNDE knockdown decreases endothelial cell proliferation, migration, and capillary tube formation. These decreases are synergistic under palmitic acid stress. These data demonstrated that lncRNA CRNDE is a regulator of endothelial cell proliferation, migration, and tube formation in response to palmitic acid, and a potential target for therapies treating the complications of obesity-induced diabetes.

LncRNA CRNDE acts as an oncogene in cervical cancer through sponging miR-183 to regulate CCNB1 expression.

Studies have identified a series of lncRNAs that contributed to various tumors, although the underlying mechanisms remain largely unclear. We proposed a ceRNA network and investigate relations among lncRNA/miRNA/mRNA in cervical cancer (CC). The genes of differential expression and lncRNA/miRNA/mRNA network were identified by combining TCGA, miRcode, starBase, miRTarBase, miRDB, TargetScan and STRING databases. Meanwhile, the function enrichment was recognized with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. Quantitative real time-PCR (qRT-PCR) was performed to determine colorectal neoplasia differentially expressed (CRNDE) expression in CC tissues and cell lines. The effects of CRNDE on the CC biological functions and cyclin B1 (CCNB1) expression were detected by conducting in vitro and in vivo experiments. Quantitative real time-PCR, western blot and dual-luciferase reporter assay were used to predict the target of miR-183. Furthermore, rescue experiments were conducted to further confirm the regulation of CCNB1 by CRNDE. Systematic analyses of bioinformatics from several databases predicted that CRNDE, miR-183 and CCNB1 were in the same network path. Their expressions were up-regulated in CC tissues and cells. Silencing CRNDE-inhibited cell proliferation, migration and invasion, restricted solid tumor growth and promoted cell apoptosis. Moreover, our results suggested that miR-183 targeted the CCNB1 3'UTR and regulated its expression. Additionally, miR-183 mimic could inverse the antitumor function of CRNDE inhibition and partially eliminated the attenuated expression of CCNB1 induced by silencing CRNDE, indicating that CRNDE could positively regulate CCNB1 expression by sponging miR-183. Our study highlighted a role for the CRNDE/miR-183/CCNB1-axis in CC and offered a promising diagnostic strategy for CC treatment.

Effect of long non-coding RNA colorectal neoplasia differentially expressed targeting microRNA-384 on invasion and migration of lung cancer SPC-A1 cells

Objective To investigate the effect of long non-coding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) targeting microRNA (miRNA, miR)-384 on invasion and migration of lung cancer SPC-A1 cells. Methods Small interfering RNA (siRNA) CRNDE was transfected into lung cancer SPC-A1 cells, and the interference effect was determined by real-time quantitative polymerase chain reaction (Real-time PCR). Cell proliferation was measured by methyl thiazol tetrazolium (MTT) assay, cell invasion and migration were measured by Transwell chamber, and Western blotting was used to detect the expression of matrix metalloproteinase (MMP)-9, MMP-2, E-cadherin and Vimentin in cells. The targeting relationship prediction software predicts that CRNDE binds to miR-384. Luciferase reporting system was used to identify the targeting relationship. In SPC-A1 cells, siRNA CRNDE and miR-384 inhibitors were co-transfected, and the above methods were used to determine the reverse effect of miR-384 inhibitors on the invasion and migration of lung cancer SPC-A1 cells transfected with siRNA CRNDE.Comparison two groups was performed using t test, and comparison three groups was performed with one-way analysis of variance (ANOVA). Results The expression of CRNDE in lung cancer SPC-A1 cells transfected with siRNA CRNDE decreased (1.01±0.12 vs. 0.43±0.06), cell proliferation decreased (0.37±0.05 vs. 0.24±0.02, F=10.467, P<0.05), the number of invasive cells (141.25±15.23 vs. 101.02±9.84, F=9.420, P<0.05) and migrating cells (188.20±16.84 vs. 142.58±11.22, F=8.345, P<0.05) decreased, the expression levels of MMP-9, MMP-2 and Vimentin proteins in cells decreased, and E-cadherin protein level increased. There were complementary binding sites between CRNDE and miR-384. CRNDE targeted and negatively regulated the expression of miR-384. The miR-384 inhibitors reversed the effects of siRNA CRNDE on proliferation (0.26±0.03 vs. 0.35±0.04, t=3.118, P<0.05), migration (139.84±10.57 vs. 176.51±12.45, t=5.276, P<0.05), invasion (99.84±10.02 vs. 134.25±11.25, t=3.890, P<0.05) and expression of MMP-9, MMP-2, E-cadherin and Vimentin in lung cancer cells. Conclusion Down-regulation of lncRNA CRNDE targeting miR-384 inhibits invasion and migration of lung cancer SPC-A1 cells. Key words: Lung cancer; MicroRNA-384; Colorectal neoplasia differentially expressed targeting; Migration; Invasion

Long non-coding RNA CRNDE enhances cervical cancer progression by suppressing PUMA expression

Cervical cancer is the second most common gynecological malignancy, and it remains a leading cause of tumor-related death among female in the world. Long non-coding RNAs (lncRNAs) have been indicated to play essential roles in tumorigenesis, and the lncRNA colorectal neoplasia differentially expressed (CRNDE) is increased in several tumors. Nevertheless, little is known about the effects of lncRNA CRNDE on human cervical cancer. The aim of the study was to explore the clinical significance of lncRNA CRNDE expression in human cervical cancer. Our results indicated that CRNDE expression was increased in cervical cancer tissues and several cervical cancer cell lines. Through loss-of-function and gain-of-function approaches, we found that CRNDE knockdown markedly reduced cervical cancer cell proliferation, while CRNDE overexpression significantly promoted cervical cancer cell growth. Consistently, CRNDE decreasing obviously inhibited tumorigenicity of cervical cancer cells in vivo, whereas CRNDE increasing markedly promoted cervical cancer progression. Mechanistically, we verified that CRNDE bond to p53 upregulated modulator of apoptosis (PUMA), and PUMA was required for CRNDE to enhance cervical cancer cell growth. Our study demonstrated that CRNDE, combined with PUMA, could be utilized as factor for the clinical diagnosis and prognosis of cervical cancer, and might be potential target for developing effective therapeutic strategy to prevent cervical cancer progression.