Long non-coding RNA LINC00174 Acts as a miR-3713 Sponge to Deregulate Oncogenic Targets, Promoting Nasopharyngeal Carcinoma Xenograft Progression via Modulating Tumor Growth, Apoptosis, and Histopathology

Description

Database Description: LINC00174-miR-3713 Axis in NPC This database curates experimental data validating lncRNA LINC00174’s oncogenic role in nasopharyngeal carcinoma (NPC) via sponging miR-3713. It integrates molecular interaction, in vitro/in vivo functional assays, and histopathology data to support NPC biomarker discovery and targeted therapy research. Core Data Modules Molecular Interaction: LINC00174 siRNA sequence (5’-GGAUUCUAGACUUCUACUATT-3’); L-KD reduces LINC00174 by 76% and increases miR-3713 by 3.8-fold (p<0.001). In Vitro Assays: C666-1 cell experiments (4 groups: Vector NC, siRNA NC, L-KD, L-KD+I). L-KD downregulates Ki-67 (84%), PCNA (90%), MMP-2 (88%), and upregulates E-cadherin (7.3-fold; p<0.001). In Vivo Tumorigenesis: BALB/c-nu/nu mice xenografts. L-KD reduces tumor volume (63%) and weight (63%) on Day28; miR-3713 inhibition partially rescues these effects (p<0.001). Histopathology: L-KD reduces nuclear pleomorphism (H&E) and increases TUNEL⁺ apoptotic cells (3.6-fold; p<0.001), reversed by miR-3713 inhibition. Utility Supports lncRNA-miRNA network analysis, NPC biomarker identification, and LINC00174-targeted therapy development. Keywords: LINC00174; miR-3713; NPC; ceRNA; therapeutic target.

Steps to reproduce

Data Generation & Reproducibility Outline This outline details standardized methods, reagents, and workflows for replicating the LINC00174-miR-3713 axis study in nasopharyngeal carcinoma (NPC): 1. Cell Culture & Transfection Cell Line: C666-1 NPC cells (STR-authenticated). Culture: RPMI-1640 (Gibco) +10% FBS (HyClone) +1% penicillin-streptomycin (Gibco), 37°C/5% CO₂. Reagents: Lipofectamine 3000 (Invitrogen); LINC00174 siRNA (5’-GGAUUCUAGACUUCUACUATT-3’, GenePharma); siRNA NC (GenePharma); miR-3713 inhibitor (50nM, RiboBio); empty vector NC (GenePharma). Groups: Vector NC, siRNA NC, L-KD (LINC00174 knockdown), L-KD+I (L-KD + miR-3713 inhibitor). Validation: qRT-PCR (48h post-transfection) with SYBR Green (Takara) to confirm LINC00174/miR-3713 expression changes. 2. In Vivo Tumorigenesis Animals: 6-week-old male BALB/c-nu/nu mice (n=3/group, Beijing Vital River), SPF housing. Xenograft: 1×10⁶ transfected cells (PBS+Matrigel 1:1, Corning) injected subcutaneously into right flank. Measurements: Tumor length/width (digital calipers, blinded) every 3 days; volume = [length×width²/2]. Sample Processing: Day28 euthanasia (cervical dislocation); tumors fixed in 4% paraformaldehyde (histology) or stored at -80°C. 3. Histopathology & Functional Assays H&E Staining: Paraffin sections (4μm) stained with hematoxylin/eosin (Sigma); scored for nuclear pleomorphism/architecture disruption (blinded pathologists). TUNEL Assay: Roche In Situ Cell Death Kit; TUNEL⁺ cells quantified via ImageJ (5 fields/section). IHC: Primary antibodies (Ki-67: Abcam ab15580; PCNA: Santa Cruz sc-56; MMP-2: Abcam ab92536; E-cadherin: CST 3195) + HRP secondary (Dako); IOD quantified via Image-Pro Plus. 4. Data Analysis Software: GraphPad Prism 8 (stats), ImageJ (cell counting), Image-Pro Plus (IOD). Stats: Student’s t-test/one-way ANOVA; significance: p<0.001. Reagents/Instruments: CO₂ incubator (Thermo Fisher), qRT-PCR system (Applied Biosystems), Olympus BX53 microscope. This outline ensures full reproducibility of the study’s findings.

Institutions

• Shanghai University

  Shanghai, Shanghai

Categories

Funders

• Medical Science Research Project of Health Commission of Hebei Province

  Grant ID: 20251470

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