lncRNA Colorectal Neoplasia Differentially Expressed Research Articles

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.

LncRNA CRNDE promotes hepatocellular carcinoma progression by upregulating SIX1 through modulating miR-337-3p.

Long noncoding RNA (lncRNA) is emerging as a vital regulator in various cancers. Recently, it was found that lncRNA colorectal neoplasia differentially expressed (CRNDE) plays an oncogenic role, promoting cell proliferation and migration in hepatocellular carcinoma (HCC). However, the underlying regulatory mechanism of lncRNA CRNDE remains unclear. The expression levels of lncRNA CRNDE and miR-337-3p were analyzed by real-time polymerase chain reaction, and sineoculis homeobox homolog 1 (SIX1) expression was determined by Western blot analysis. RNA pull-down, luciferase and Western blot analysis assays were used to examine the target relationship between lncRNA CRNDE and miR-337-3p as well as between miR-337-3p and SIX1. The functional effects of lncRNA CRNDE and miR-337-3p were examined in vitro by using cell viability, colony formation, wound scratch, transwell assays, and in vivo in a xenograft tumor mouse model. LncRNA CRNDE was overexpressed in tumor tissues of patients with HCC. LncRNA CRNDE downregulation significantly suppressed cell proliferation and migration. Mechanistic investigations demonstrated that lncRNA CRNDE interacted with miR-337-3p and decreased its expression, thereby increasing the protein expression of miR-337-3p's target, SIX1. In addition, in vivo experiments using a xenograft tumor mouse model revealed that lncRNA CRNDE served as an oncogene, partly through sponging miR-337-3p and upregulating SIX1 in HCC. In this study, we report a newly identified regulatory mechanism lncRNA CRNDE/miR-337-3p/SIX1 axis, suggesting a promising therapeutic target in HCC.

The role of lncRNA CRNDE in obesity‐associated endothelial dysfunction

Obesity, a risk factor for type II diabetes and cardiovascular disease affects 35.7% of the U.S. population and costs approximately $210 billion per year. Endothelial cells (ECs) are cells that line blood vessels, are important in signaling and transport and have been implicated in obesity. The role of long non‐coding RNA (lncRNA) in regulating endothelial function, especially under disease conditions, has been understudied. In an RNA‐seq analysis of palmitate‐treated ECs, the lncRNA Colorectal Neoplasia Differentially Expressed (CRNDE) was determined to be upregulated. The purpose of this study is to determine the role of CRNDE in EC function through examination of various signaling pathways and exploring how obesogenic stimuli induce CRNDE expression. To determine the effect of obesogenic stimuli on CRNDE expression, HUVECs were treated with albumin‐conjugated palmitic acids or albumin for 4 or 12 hours and CRNDE expression was determined by qPCR. CRNDE is significantly increased in palmitic acid‐treated HUVECs after 12 hours compared with cells treated with albumin. Bioinformatic analysis of CRNDE and its promoter reveals the presence of p53 binding sites at CRNDE promoter which mediate the induction of CRNDE. To test this, CRNDE expression will be examined in HUVECs in the presence or absence of p53 knockdown. Chromatin immunoprecipitation will be used to determine the binding of p53 at the CRNDE promoter in HUVECs. Loss‐of‐ and gain‐of‐function studies will be performed to examine the signaling pathway that is regulated by CRNDE. Understanding the role of CRNDE in obesity‐associated endothelial dysfunction could provide new targets to reduce vascular complications associated with obesity and diabetes.Support or Funding InformationUCAREThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

LncRNA CRNDE promotes hepatocellular carcinoma cell proliferation, invasion, and migration through regulating miR-203/ BCAT1 axis.

To investigate the impact of long noncodingRNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) on hepatocellular cancer (HCC) cell propagation, invasion, and migration by mediating miR-203/ BCAT1 axis. Microarray analysis was based on 25 pairs of HCC cancerous tissues and adjacent tissues. The expression levels of CRNDE, miR-203, and BCAT1 in HCC tissues were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The liver cell line L-02 and HCC cell lines HepG2 and Huh-7 were utilized to assess the regulatory effects of CRNDE and miR-203 on HCC progression in vitro. Western blot was used to qualify BCAT1 protein expression level. Cell proliferation and apoptosis were evaluated using CCK-8 and flow cytometry analysis, whereas cell invasion and migration assay were performed by the Transwell assay. The relationship among CRNDE, miR-203, and BCAT1 was validated by dual luciferase assay. Tumor Xenograft study was established to verify the pathological effect of CRNDE on HCC development in vivo. The expression levels of the CRNDE and BCAT1 were upregulated in HCC tissues and cells, whereas miR-203 was downregulated in HCC. Knockdown of CRNDE or miR-203 overexpression would inhibit HCC cell propagation and metastasis, and induced cell apoptosis. Moreover, miR-203 was negatively correlated with CRNDE, the same as miR-203 with BCAT1. Dual luciferase assay showed that miR-203 was an inhibitory target of CRNDE, and BCAT1 was directly targeted by miR-203 as well. LncRNA CRNDE could enhance HCC tumorgenesis by sponging miR-203 and mediating BCAT1. LncRNA CRNDE might facilitate HCC cell propagation, invasiveness, and migration through regulating miR-203/ BCAT1 axis.

LncRNA CRNDE promotes the epithelial-mesenchymal transition of hepatocellular carcinoma cells via enhancing the Wnt/β-catenin signaling pathway.

Colorectal neoplasia differentially expressed (CRNDE) is a significantly upregulated long noncoding RNA in hepatocellular carcinoma (HCC). CRNDE could promote cell proliferation, migration, and invasion, while its molecular mechanisms were still largely unclear. In this study, we investigated the expression and function of CRNDE. CRNDE was significantly upregulated in tumor tissues compared with adjacent normal tissues. In vitro, we revealed that knockdown of CRNDE inhibited cell proliferation, migration, and cell invasion capacities in HCC. Animal studies indicated that CRNDE knockdown represses both growth and metastasis of HCC tumors in vivo. Moreover, knockdown of CRNDE suppressed the cell epithelial‐mesenchymal transition (EMT) process by increasing the expression of E‐cadherin and ZO‐1, whereas, decreasing the expression of N‐cadherin, slug, twist, and vimentin in HCC cells. We also revealed that knockdown of CRNDE suppressed the Wnt/β‐catenin signaling in HCC. Thus, CRNDE could modulate EMT of HCC cells and knockdown of CRNDE impaired the mesenchymal properties. CRNDE increased invasion of HCC cells might be through activating the Wnt/β‐catenin signaling pathway.

The long non-coding RNA CRNDE competed endogenously with miR-205 to promote proliferation and metastasis of melanoma cells by targeting CCL18

ABSTRACTMelanoma was the most malignant skin neoplasm with an increasing morbidity around the world. Although new immunotherapies and targeted therapies have emerged recently, the long-term survival of melanoma patients still remains low. To reveal effective diagnostic methods and therapeutic strategies, the potential mechanism of melanoma is urgently needed to be studied. Long non-coding RNAs (lncRNAs) have become an important regulatory factor in the occurrence and development of cancer, and it can be used as a new prognostic and diagnostic marker. In this study, we aimed to inspect the effects of lncRNA colorectal neoplasia differentially expressed (CRNDE) on the melanoma cell viability, invasion and migration. After microarray analysis, 106 dysregulated lncRNAs and 1187 abnormally expressed mRNAs were screened out. Further, the lncRNA CRNDE and CCL18 expression in melanoma tissues and cell lines were examined. It was determined that they were both overexpressed in melanoma tissues and cell lines. The down-regulation of lncRNA CRNDE and CCL18 induced melanoma cell apoptosis and inhibited cell viability. Then, miR-205 which had binding site with lncRNA CRNDE and CCL18 was involved in the next experiment, and it was down-regulated in melanoma that negatively correlated with lncRNA CRNDE expression. In addition, overexpression of miR-205 results in the restore of cell viability and aggressiveness. In conclusion, LncRNA CRNDE promotes the migration and invasion of melanoma by sponging miR-205 and releasing CCL18.

LncRNA CRNDE is a biomarker for clinical progression and poor prognosis in clear cell renal cell carcinoma.

Colorectal neoplasia differentially expressed (CRNDE) served as an oncogenic long noncoding RNA (lncRNA) to be involved in the initialization and development of human cancers. However, the clinical significance and biological function of CRNDE in clear cell renal cell carcinoma (ccRCC) was not fully understood. In our study, we found CRNDE levels were increased in ccRCC tissue specimens and cell lines, and corrected with advanced clinical stage, large tumor size, lymph node metastasis, distant metastasis, and poor pathological grade in patients with ccRCC. Furthermore, levels of CRNDE were negatively correlated with overall survival of patients with ccRCC, and high-expression of CRNDE was an independent poor prognostic factor for patients with ccRCC. Moreover, loss-of-function and gain-of-function approaches showed CRNDE-enhanced ccRCC cell migration and invasion through modulating EMT-associated genes. In conclusion, CRNDE acts as an oncogenic lncRNA in ccRCC.

LncRNA CRNDE promotes cell proliferation, invasion and migration by competitively binding miR-384 in papillary thyroid cancer.

Thyroid cancer is one of the most prevalent endocrine neoplasm. The present study examined the effects of Colorectal Neoplasia Differentially Expressed (CRNDE) on the progression of papillary thyroid cancer (PTC), and explored the underlying molecular mechanisms. Quantitative real-time PCR was used to detect CRNDE, miR-384 and pleiotrophin (PTN) mRNA expression. Western blot was used to measure PTN protein levels. Cell proliferation, cell growth, cell invasion and migration of PTC cells were determined by CCK-8, colony formation, transwell invasion and migration assays, respectively. CRNDE was up-regulated in PTC tissues and cell lines. Overexpression of CRNDE promoted BCPAP cell proliferation, invasion and migration, while knock-down of CRNDE suppressed K1 cell proliferation, invasion and migration. CRNDE negatively regulated the expression of miR-384 in PTC cells, which was further confirmed by luciferase reporter assay. MiR-384 was down-regulated and inversely correlated with CRNDE expression in PTC tissues. MiR-384 suppressed cell proliferation, invasion and migration in PTC cells, and enforced expression of miR-384 attenuated the oncogenic effects of CRNDE in PTC cells. PTN was predicted as a downstream target of miR-384, which was confirmed by luciferase reporter assay, and PTN was up-regulated in PTC tissues, and was negatively correlated with miR-384 expression and positively correlated with CRNDE expression in PTC tissues. In summary, our results suggested that the CRNDE/miR-384/PTN axis may play an important role in the regulation of PTC progression, which provides us with new insights into understanding the PTC.

Long Noncoding RNA CRNDE Promotes Multiple Myeloma Cell Growth by Suppressing miR-451.

It has been determined that long noncoding RNAs (lncRNAs) are identified as a potential regulatory factor in multiple tumors as well as multiple myeloma (MM). However, the role of colorectal neoplasia differentially expressed (CRNDE) in the pathogenesis of MM remains unclear. In this study, we found that the CRNDE expression level, in MM samples and cell lines, is higher than that in the control detected by real-time qPCR, which is also closely related to tumor progression and poor survival in MM patients. Knockdown of CRNDE significantly inhibits the proliferative vitality of MM cells (U266 and RPMI-8226), induces cell cycle arrest in the G0/G1 phase, and promotes apoptosis. After being transfected with siRNA, miR-451 expression observably increases. Bioinformatics analysis and luciferase assay reveal the interaction by complementary bonding between CRNDE and miR-451. Pearson’s correlation shows that CRNDE is negatively correlated to miR-451 expression in human MM samples. Subsequently, miR-451 inhibitor rescues the inhibited tumorigenesis induced by CRNDE knockdown. Our study illustrates that lncRNA CRNDE induces the proliferation and antiapoptosis capability of MM by acting as a ceRNA or molecular sponge via negatively targeting miR-451, which could act as a novel diagnostic marker and therapeutic target for MM.

Long noncoding RNA CRNDE promotes colorectal cancer cell proliferation via epigenetically silencing DUSP5/CDKN1A expression

Evidence indicates that long non-coding RNAs (lncRNAs) play a critical role in the regulation of tumor cellular processes, such as proliferation, apoptosis, and metastasis. LncRNA CRNDE (Colorectal Neoplasia Differentially Expressed) is located at human chromosome 16 and has been found overexpressed in a variety of cancers including colorectal cancer (CRC). In this paper, we report that lncRNA CRNDE expression was remarkably upregulated in CRC tissues and that lncRNA CRNDE overexpression was positively correlated with advanced pathological stages and larger tumor sizes. In addition, the knockdown of CRNDE significantly suppressed proliferation and caused apoptosis of CRC cells both in vitro and in vivo. Furthermore, RNA immunoprecipitation and chromatin immunoprecipitation assays demonstrated that lncRNA CRNDE could epigenetically suppress the expressions of dual-specificity phosphatase 5 (DUSP5) and CDKN1A by binding to EZH2 (the key components of Polycomb repressive complex 2 (PRC2)), thus promoting CRC development. In conclusion, our data suggest that the lncRNA CRNDE promotes the progression of CRC and is a potential therapeutic target for CRC intervention.

Analysis of long non-coding RNA expression profiles in clear cell renal cell carcinoma.

To investigate the expression patterns of long non-coding RNAs (lncRNAs) in clear cell renal cell carcinoma (ccRCC) and in metastatic renal cell carcinoma (RCC), the present study downloaded three human exon arrays available from the public Gene Expression Omnibus. The probes of the human exon arrays were re-annotated and the probes uniquely mapping to lncRNAs were retained at the gene level. Following the analysis of GSE53757 and GSE46699, which contained paired ccRCC cancer and normal adjacent tissue samples, 32 differentially expressed lncRNAs (adjusted P<0.01) in ccRCC were identified. Various lncRNAs, including ENSG00000177133, NR_024418, T-cell leukemia/lymphoma 6 (TCL6), growth arrest-specific transcript 5, deleted in lymphocytic leukemia 2, colorectal neoplasia differentially expressed (CRNDE) and MIR155HG, have been reported to be abnormally expressed in cancers. Of these genes, NR_24418 and TCL6 have been reported to be associated with ccRCC. Following analysis of GSE47352, which contained 4 primary metastatic and 5 non-metastatic tumor samples, the 50 top differentially expressed lncRNAs were identified in metastatic ccRCC (Mann-Whitney U test, P<0.05). Comparison with the ccRCC associated lncRNAs revealed that the lncRNA CRNDE demonstrated an increased expression in ccRCC and metastatic ccRCC samples, which suggested that CRNDE is important in the progression of ccRCC. The lncRNA ENSG00000244020 was decreased in ccRCC and metastatic ccRCC, suggesting that silencing of ENSG00000244020 may be important in ccRCC development. Overall, a set of lncRNAs was identified as differentially expressed in ccRCC and metastatic ccRCC, providing potential candidates for the discovery of novel cancer biomarkers and therapeutic targets to improve diagnosis and therapy in RCC.

Long noncoding RNA CRNDE stabilized by hnRNPUL2 accelerates cell proliferation and migration in colorectal carcinoma via activating Ras/MAPK signaling pathways

Recent studies have furthered our understanding of the function of long noncoding RNAs (lncRNAs) in numerous biological processes, including cancer. This study investigated the expression of a novel lncRNA, colorectal neoplasia differentially expressed (CRNDE), in colorectal carcinoma (CRC) tissues and cells by real-time RT-PCR and in situ hybridization, and its biological function using a series of in vitro and in vivo experiments to determine its potential as a prognostic marker and therapeutic target. CRNDE was found to be upregulated in primary CRC tissues and cells (P<0.05), and the upregulation of CRNDE expression is a powerful predictor of advanced TNM stage (P<0.05) and poor prognosis for CRC patients (P=0.002). The promoting effects of CRNDE on the cell proliferation, cell cycling and metastasis of CRC cells were confirmed both in vitro and in vivo by gain-of-function and loss-of-function experiments. Mechanistically, it was demonstrated that CRNDE could form a functional complex with heterogeneous nuclear ribonucleoprotein U-like 2 protein (hnRNPUL2) and direct the transport of hnRNPUL2 between the nucleus and cytoplasm. hnRNPUL2 that was accumulated in the cytoplasm could interact with CRNDE both physically and functionally, increasing the stability of CRNDE RNA. Moreover, gene expression profile data showed that CRNDE depletion in cells downregulated a series of genes involved in the Ras/mitogen-activated protein kinase signaling pathways. Collectively, these findings provide novel insights into the function and mechanism of lncRNA CRNDE in the pathogenesis of CRC and highlight its potential as a therapeutic target for CRC intervention.

LncRNA CRNDE triggers inflammation through the TLR3-NF-κB-Cytokine signaling pathway.

Colorectal neoplasia differentially expressed (CRNDE), an oncogene, is highly expressed in many tumor cells and affects cellular proliferation, migration, invasion, and apoptosis. Its function and mechanism of action is a research hotspot. In this study, microarray analysis was performed to discover the differentially expressed genes in CRNDE over-expression cells. RT² Profiler PCR Array was used to study the expression of genes related to the toll-like receptor (TLR) pathway. We found that over-expression of CRNDE in astrocytes increased the expression of key factors in the toll-like receptor signaling pathway, especially toll-like receptor-3-mediated MyD88-independent pathway. Furthermore, it up-regulated expression levels of downstream transcription factor such as nuclear factor kappa B and numerous cytokines. In contrast, CRNDE knockdown in glioma U87MG cell line showed an opposite trend in the expression of the above-mentioned genes. We speculated that CRNDE might trigger inflammation to regulate tumorigenesis and tumor development through the toll-like receptor pathway.

Highly expressed lncRNA CRNDE promotes cell proliferation through Wnt/β-catenin signaling in renal cell carcinoma.

Recently, numerous studies revealed that long non-coding RNAs (lncRNAs) play complex roles in the field of tumor biology, while the functions of lncRNA in renal cell carcinoma (RCC) remain largely unknown. In the current study, we retrieved Oncomine database and found a lncRNA colorectal neoplasia differentially expressed (CRNDE) which is highly expressed in different cohorts of RCC patients; this clue reminds us that CRNDE might exert its functions in RCC tumorigenesis. We then detected the level of CRNDE in fresh RCC tissues and found that CRNDE is significantly up-regulated compared with adjacent tissues. Furthermore, both loss and gain function assays revealed that CRNDE promotes RCC cell proliferation and growth both in vitro and in vivo.In addition, we found that CRNDE regulates the cell cycle transition from G0/G1 stage to S stage and modulates the expression of CCND1 and CCNE1. Moreover, we further illustrated that CRNDE activates Wnt/β-catenin signaling in RCC cell lines. Taken together, in the current study, we found that lncRNA CRNDE is highly expressed in RCC malignant tissues and the heightened CRNDE robustly promotes RCC cell proliferation through activating Wnt/β-catenin signaling; our findings enlarge our knowledge in the molecular pathology of RCC tumorigenesis.