Association of Serum lncRNA CASC11 with Injury Severity and Inflammation in Spinal Cord Injury

Summary

In spinal cord injury (SCI) patients, CASC11 upregulation is associated with injury severity. In vitro, CASC11 downregulation alleviated apoptosis and inflammation, suggesting an association with the miR-130b-5p/SPP1 axis. These results suggest CASC11’s potential role as a biomarker candidate in SCI pathogenesis.

Abstract

Spinal cord injury (SCI) is a severe central nervous system trauma. This single-center, observational and in vitro study investigated the diagnostic potential of CASC11 and its possible regulatory mechanism in SCI using clinical serum samples and lipopolysaccharide-stimulated cell models. CASC11, miR-130b-5p, and SPP1 levels were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Cellular functions and targeting relationships were assessed via cell counting kit-8 (CCK-8), flow cytometry, western blot, enzyme-linked immunosorbent assay (ELISA), dual-luciferase reporter, and RNA immunoprecipitation (RIP) assays. CASC11 was highly expressed in SCI and associated with inflammation as a potential diagnostic biomarker. In lipopolysaccharide (LPS)-treated PC-12 cells, silencing CASC11 alleviated suppressed cell activity, apoptosis, and inflammation, which was reversed by miR-130b-5p inhibitor. These findings suggest that CASC11 is associated with SCI severity, while in vitro data indicate its involvement in inflammatory and apoptotic responses through the CASC11/miR-130b-5p/SPP1 axis. This study is limited by its single-center design and lack of in vivo validation.

Introduction

Spinal cord injury (SCI) denotes a temporary or permanent systemic disorder arising from spinal cord damage caused by external forces or diseases, resulting in impairment of motor, sensory, reflex, and autonomic nervous system functions^1,2. Patients frequently encounter lifelong physical functional deficits and complications, imposing considerable suffering and burdens on individuals and families. The etiology of SCI is broadly classified into traumatic and non-traumatic origins³. Traumatic injuries, predominantly resulting from spinal fractures due to external forces such as traffic ac....

Protocol

Enrollment of subjects

Patients with acute spinal trauma (n = 300) treated in the hospital from January 2023 to May 2025 were selected and classified into the complete spinal cord injury (CSCI) group (grade A, n = 98), incomplete spinal cord injury (ISCI) group (grades B, C, and D; n = 95), and normal neurological function (NNF) group (grade E, n = 107) according to the American Spinal Injury Association Impairment Scale (AIS classification). For patients in the NNF group, radiological evidence of SCI, such as edema, contusion, or compression, was confirmed by magnetic resonance imaging (MRI), but no sensory or motor defici....

Results

Basic data of the subjects

The baseline characteristics of patients in the Control, NNF, ISCI, and CSCI groups are presented in Table 1. Statistical analysis revealed no significant inter-group differences in variables, such as sex, age, body mass index (BMI), etiology of injury, and time of injury (P > 0.05). Significant differences among the four groups were observed for length of stay, ASIA grade, and the concentrations of the pro-inflammatory c.......

Discussion

SCI is a devastating condition of the central nervous system that frequently results in significant deficits in sensation, motor control, and autonomic function, thus substantially diminishing a patient’s quality of life and long-term prognosis¹⁸. Patients may present with functional impairment and sensory loss following spinal trauma, and imaging studies (such as MRI) can be employed to assess the occurrence of SCI¹⁹. The onset of SCI triggers a deleterious secondary.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
BCA protein assay kit	Yeasen, China	P0010
CCK-8 solution	Sigma-Aldrich, USA	96992
Dual-luciferase reporter vector	Promega, USA	E133A	pmirGLO vector
Dulbecco’s Modified Eagle Medium	ATCC, Manassas, VA, USA	30-2002
ELISA kit for IL-1β	R&D Systems, USA	DY401
ELISA kit for IL-6	R&D Systems, USA	HS600B
ELISA kit for TNF-α	R&D Systems, USA	DTA00C
Fetal bovine serum	Gibco, Grand Island, NY, USA	10099141
Flow cytometer	BD FACSCalibur, USA	FACS101
Image analysis software	NIH, Bethesda, MD, USA	version 1.44p	ImageJ software
Isopropanol	Beijing Chemical Factory, China	32064
Lipid-based transfection reagent	Invitrogen, Carlsbad, CA, USA	L3000008	Lipofectamine 3000
Lipopolysaccharide	Sigma-Aldrich, USA	L2880
Lysis buffer	Beyotime, China	P0013D	RIPA lysis buffer
Microplate reader	Reagen, China	RNE90002
PC-12 and BV-2 cells	ATCC, Manassas, USA
Phenol-chloroform reagent	Thermo, USA	15596026	Trizol reagent
Reverse transcription kit	Takara, Japan	RR047A	PrimeScript RT Kit
RNA-binding protein immunoprecipitation kit	Millipore, USA	17-700	Magna RIP Kit
RNase-free microtubes	Axygen, Corning, NY, USA	MCT-150-C-S
Serum separator tubes	Terumo, Tokyo, Japan	VP-AS109K60
Statistical analysis software	San Diego, USA	version 9.0	GraphPad Prism
SYBR Green premix	Takara, Japan	RR420A	SYBR Premix Ex Taq