Dataset Information

Long Non-coding RNA PVT1 Competitively Binds MicroRNA-424-5p to Regulate CARM1 in Radiosensitivity of Non-Small-Cell Lung Cancer.

ABSTRACT: Accumulating evidence revealed that dysregulated long non-coding RNAs (lncRNAs) were involved in tumorigenesis and progression. This study is supposed to reveal the effects of lncRNA PVT1 on the radiosensitivity of non-small-cell lung cancer (NSCLC) via the microRNA (miR)-424-5p/lncRNA PVT1/CARM1 signaling pathway. Differentially expressed lncRNA was filtrated. The co-expressed gene of lncRNA was predicted, and gene ontology analysis was performed to find out the genes associated with NSCLC radiosensitivity. The miR that was combined with lncRNA and mRNA was filtrated. Two cell lines with the highest expressed PVT1 were selected, followed by transfection with a series of different mimic, inhibitor, or siRNA. RIP assay was employed for the interaction between PVT1 and CARM1. The regulatory effect of miR-424-5p on cell proliferation, migration, invasion, cycle, and apoptosis was investigated. PVT1 was the most remarkable lncRNA that upregulated in NSCLC. CARM1 co-expressed with lncRNA PVT1 and associated with NSCLC radiosensitivity. Both lncRNA PVT1 and CARM1 can combine with miR-424-5p. Increased PVT1, CARM1, MMP-2, MMP-9, and Bcl-2 and decreased miR-424-5p and Bax were found in NSCLC tissues. PVT1 was targeted by miR-424-5p. After silencing of PVT1 or overexpressed miR-424-5p, decreased PVT1, CARM1, MMP-2, MMP-9, and Bcl-2 inhibited cell proliferation, migration, and invasion but promoted miR-424-5p, Bax, and cell apoptosis. The present study confirms the radiosensitivity of NSCLC radiotherapy can be increased by siRNA-PVT1 and overexpressed miR-424-5p.

SUBMITTER: Wang D

PROVIDER: S-EPMC6411630 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Long Non-coding RNA PVT1 Competitively Binds MicroRNA-424-5p to Regulate CARM1 in Radiosensitivity of Non-Small-Cell Lung Cancer.

Wang Dong D Hu Yi Y

Molecular therapy. Nucleic acids 20181215

Accumulating evidence revealed that dysregulated long non-coding RNAs (lncRNAs) were involved in tumorigenesis and progression. This study is supposed to reveal the effects of lncRNA PVT1 on the radiosensitivity of non-small-cell lung cancer (NSCLC) via the microRNA (miR)-424-5p/lncRNA PVT1/CARM1 signaling pathway. Differentially expressed lncRNA was filtrated. The co-expressed gene of lncRNA was predicted, and gene ontology analysis was performed to find out the genes associated with NSCLC radi ...[more]

PMID: 30861415

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Project description:Purpose:Long intergenic non-coding RNA 922 (LINC00922) plays a critical role in the progression of lung cancer. In this study, we aimed to quantify LINC00922 expression in breast cancer and determine its influence on the malignant behavior of breast cancer cells in vitro and in vivo. We also investigated the mechanism by which LINC00922 affects the progression of breast cancer. Materials and Methods:Reverse transcription-quantitative polymerase chain reaction was performed to quantify LINC00922 expression in breast cancer tissues and cell lines. The cell counting kit-8 assay, flow cytometry, Transwell migration and invasion assays, and tumor model assays were performed to determine the effects of LINC00922 deficiency on breast cancer cell proliferation, apoptosis, migration and invasion in vitro, and tumor growth in vivo, respectively. Furthermore, bioinformatics analysis was performed to predict the potential target microRNA of LINC00922. The prediction was further evaluated using luciferase reporter and RNA immunoprecipitation assays. Results:LINC00922 was clearly overexpressed in breast cancer tissues and cell lines. LINC00922 depletion restricted breast cancer cell proliferation, migration, and invasion but induced cell apoptosis in vitro. Additionally, its knockdown evidently repressed tumor growth of breast cancer cells in vivo. Mechanistically, LINC00922 was demonstrated to serve as a molecular sponge for miR-424-5p in breast cancer cells. Furthermore, brain-derived neurotrophic factor (BDNF) was verified as a direct target of miR-424-5p in breast cancer cells, and BDNF expression was found to be positively regulated by LINC00922 through sponging miR-425-5p. Rescue experiments further revealed that the influences on breast cancer cell proliferation, apoptosis, migration, and invasion induced by LINC00922 silencing were abrogated by increasing the output of the miR-424-5p/BDNF axis. Conclusion:The LINC00922/miR-424-5p/BDNF pathway is implicated in the acceleration of the malignant behavior of breast cancer cells. These findings suggest that this pathway is a promising novel molecular target in breast cancer therapy.

Project description:Previous studies have demonstrated that long non‑coding RNAs (lncRNAs) serve a key role in the development and progression of several types of cancer, including glioma. The lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) contributes to cancer growth through its effects on cell proliferation, migration, invasion and drug resistance. However, the exact regulatory mechanisms via which NEAT1 acts in glioma are unclear. In the present study, the expression levels and function of NEAT1 in glioma tissues and cell lines were examined in vitro and in vivo. By reverse transcription‑quantitative PCR and fluorescence in situ hybridization analysis, NEAT1 expression was upregulated in glioma tissues compared with in adjacent normal brain tissues, and elevated NEAT1 levels were associated with poor prognosis. Cell Counting Kit‑8, colony formation, ethynyldeoxyuridine, flow cytometry and western blotting assays were performed to detect the effects of NEAT1 on cell biological behavior. Knockdown of NEAT1 in glioma cell lines was associated with cell cycle arrest at the G0/G1 phase, decreased proliferation and elevated apoptosis in vitro, and resulted in reduced tumor growth and increased survival in a mouse xenograft model of glioma. Using bioinformatics analysis, RNA immunoprecipitation experiments and luciferase reporter assays, it was demonstrated that NEAT1 may competitively bind to microRNA (miR)‑324‑5p, thus blocking its interaction with target mRNAs. Potassium channel tetramerization protein domain containing 20 (KCTD20) was identified as a specific miR‑324‑5p target. Accordingly, the inhibition of NEAT1 resulted in the downregulation of KCTD20 through competitive binding with miR‑324‑5p, decreased cell proliferation and increased apoptosis. Concomitant NEAT1 knockdown and inhibition of miR‑324‑5p partially reversed the effects of NEAT1 knockdown on cell proliferation and apoptosis, and further regulated KCTD20 expression. Collectively, the present findings demonstrated that NEAT1 acted as a competing endogenous RNA for miR‑324‑5p, and identified the NEAT1/miR‑324‑5p/KCTD20 axis as a novel regulatory axis and a potential therapeutic target for human glioma.

Project description:BackgroundAcquired drug-resistance is the major risk factor for poor prognosis and short-term survival in patients with osteosarcoma (OS). Accumulating evidence has revealed that long noncoding RNAs (lncRNAs), including plasmacytoma variant translocation 1 (PVT1), play potential regulatory roles in the malignant development of OS. Considering the subcellular distribution of PVT1 as both nuclear and cytoplasmic lncRNA, a thorough exploration of its extensive mechanisms becomes essential.MethodsThe GEO database was utilized for the acquisition of gene expression data, which was subsequently analyzed to fulfill the research objectives. The subcellular localization of PVT1 in OS cells was determined using fluorescence in situ hybridization (FISH) and quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, the sensitivity of OS cells to doxorubicin was comprehensively evaluated through measurements of cell viability, site-specific proliferation capacity, and the relative expression abundance of multidrug resistance-related proteins (MRPs). In order to investigate the differential response of OS cells with varying levels of PVT1 expression to doxorubicin, pulmonary metastasis mice models were established for in vivo studies. Molecular interactions were further examined using the dual-luciferase assay and RNA immunoprecipitation assay (RIP) to analyze the binding sites of miR-15a-5p and miR-15b-5p on PVT1 and G1/S-specific cyclinD1 (CCND1) mRNA. Furthermore, the chromatin immunoprecipitation (ChIP) and dual-luciferase assay were employed to assess the transcriptional activation of the proto-oncogene c-myc (MYC) on the CCND1 promoter and identify the corresponding binding sites.ResultsIn doxorubicin resistant OS cells, transcription levels of PVT1, MYC and CCND1 were significantly higher than those in original cells. In vivo experiments demonstrated that OS cells rich in PVT1 expression exhibited enhanced tumorigenicity and resistance to doxorubicin. In vitro experiments, it has been observed that overexpression of PVT1 in OS cells is accompanied by an upregulation of CCND1, thereby facilitating resistance to doxorubicin. Nonetheless, this PVT1-induced resistance can be effectively attenuated by the knockdown of CCND1. Mechanistically, PVT1 functions as a competitive endogenous RNA (ceRNA) that influences the expression of CCND1 by inhibiting the degradation function of miR-15a-5p and miR-15b-5p on CCND1 mRNA. Additionally, as a neighboring gene of MYC, PVT1 plays a role in maintaining MYC protein stability, which further enhances MYC-dependent CCND1 transcriptional activity.ConclusionThe resistance of OS cells to doxorubicin is facilitated by PVT1, which enhances the expression of CCND1 through a dual mechanism. This findings offer a novel perspective for comprehending the intricate regulatory mechanisms of long non-coding RNA in influencing the expression of coding genes.

Project description:Background:A long non-coding RNA termed as long intergenic non-protein coding RNA 491 (LINC00491) has been validated as an oncogene to promote cancer progression in colon adenocarcinoma. The goal of this study was to determine the expression and carcinogenic functions of LINC00491 in non-small-cell lung cancer (NSCLC). Besides, it was aimed to understand how LINC00491 affects the malignant processes of NSCLC cells. Methods:The expression of LINC00491 in NSCLC was investigated by bioinformatic analysis and reverse transcription-quantitative PCR. After LINC00491 knockdown, cell counting kit-8 assay, flow cytometry, migration and invasion detection assays as well as nude mice xenograft assay were conducted to test the roles of LINC00491 in NSCLC cells. Two online databases, StarBase 3.0 and miRDB, were utilized to determine the putative target miRNA of LINC00491, and the prediction was subsequently confirmed by luciferase reporter assay, RNA immunoprecipitation assay, reverse transcription-quantitative PCR, Western blotting, and rescue assays. Results:LINC00491 was overexpressed in both NSCLC tissues and cell lines. Functional investigation revealed that depleted LINC00491 facilitated cell apoptosis and decreased cell proliferation, migration, and invasion in vitro. Additionally, the downregulation of LINC00491 impaired NSCLC cell tumor growth in vivo. Mechanistically, LINC00491 functioned as a competing endogenous RNA by sponging microRNA-324-5p (miR-324-5p) in NSCLC cells. miR-324-5p was weakly expressed in NSCLC and exerted tumor-suppressing actions during cancer progression. Furthermore, specificity protein 1 (SP1) was validated as the direct target of miR-324-5p in NSCLC and was under the regulation of LINC00491 via sponging miR-324-5p. Rescue experiments reconfirmed that miR-324-5p inhibition and SP1 overexpression both abrogated the suppressive roles of LINC00491 deficiency in NSCLC cells. Conclusion:LINC00491 promoted the oncogenicity of NSCLC via serving as a miR-324-5p sponge, which further upregulated the expression of SP1. The LINC00491/miR-324-5p/SP1 pathway disclosed a new mechanism of NSCLC pathogenesis and may provide effective targets for better NSCLC treatment.

Dataset Information

Long Non-coding RNA PVT1 Competitively Binds MicroRNA-424-5p to Regulate CARM1 in Radiosensitivity of Non-Small-Cell Lung Cancer.

Publications

Long Non-coding RNA PVT1 Competitively Binds MicroRNA-424-5p to Regulate CARM1 in Radiosensitivity of Non-Small-Cell Lung Cancer.

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