BMC Molecular andCell Biology(2023) 24:15
Jiang et al. BMC Molecular and Cell Biology
https://doi.org/10.1186/s12860-023-00477-2

Open Access

RESEARCH

SCAT8/miR‑125b‑5p axis triggers malignant
progression of nasopharyngeal carcinoma
through SCARB1
Chunmao Jiang1, Dandan Feng2, Yu Zhang3, Kun Yang4, Xiaotong Hu1 and Qian Xie1*

Abstract
Nasopharyngeal carcinoma is a tumor with high malignancy and poor prognosis, which severely affects the health ofthe patients. LncRNAs and microRNAs are crucial for the occurrence and development of nasopharyngeal carcinoma,
which regulate the progression of nasopharyngeal carcinoma through the ceRNA network. SCARB1 plays an essential
role in nasopharyngeal carcinoma. However, the mechanism underlying the regulation of SCARB1 in nasopharyngeal
carcinoma through non-coding RNAs remains unclear. Our findings indicated that the SCAT8/miR-125b-5p axis promoted the malignant progression of nasopharyngeal carcinoma by driving the expression of SCARB1. Mechanistically,
the expression of SCARB1 could be regulated by the lncRNA, SCAT8 and the microRNA, miR-125b-5p. Moreover, as aceRNA of miR-125b-5p, SCAT8 can not only regulate the expression of SCARB1, but also regulate the malignant progression of nasopharyngeal carcinoma. Notably, our results reveal a novel ceRNA regulatory network in nasopharyngeal carcinoma, which could serve as a potential target for the diagnosis and treatment of nasopharyngeal carcinoma.
Keywords Nasopharyngeal carcinoma, ceRNA, SCARB1, Cell proliferation, Cell migration
Introduction
Nasopharyngeal carcinoma occurs in the nasopharynx,
nasopharyngeal cavity, and upper throat, and is common
across Southeast Asia and Africa [28]. The position of thenasopharynx in the center of the head, close to the baseof the skull, with significant blood vessels and nerves
passing nearby, makes it highly susceptible to cervicallymph node metastases [34]. Early-stage patients have*Correspondence:
Qian Xie
x477634153@163.com
1
Department of Health Management, Daping Hospital, Army Medical
University, Chongqing 400010, China
2
Department of Otolaryngology Head and Neck Surgery, Daping
Hospital, Army Medical University, Chongqing 400010, China
3
Department of Otolaryngology Head and Neck Surgery, the First
Affiliated Hospital of Chongqing Medical University, Chongqing 400010,
China
4
Department of Health Management Center, the Second AffiliatedHospital of Chongqing Medical University, Chongqing 400010, China

a cure rate of more than 80%, but nasopharyngeal carcinoma in the early stage is difficult to detect due to thelocation of the nasopharynx [30]. More than 70% of thepatients see a doctor regularly for common symptoms
like neck lumps and nasal congestion, which are similar to those of nasopharyngeal cancer [33]. In the earlystages, radiotherapy alone shows excellent results but
patients with an advanced or recurrent disease may need
a combination of chemotherapy and surgery [14]. After
regular treatment, the average five-year survival rateis nearly 60% and more than 30% for advanced patients[37]. Scientists worldwide have been searching for effective treatment strategies to reduce the recurrence rate ofnasopharyngeal carcinoma and improve patient survival.
Non-coding RNA (ncRNA) refers to RNA without
protein-coding ability, and mainly includes circular RNA(circRNA), long non-coding RNA (lncRNA), and micro
RNA (miRNA), among other types [12, 32]. Because ncRNAs are closely related to human diseases, their study© The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which
permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to theoriginal author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images orother third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line
to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory
regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of thislicence, visit http://​creat​iveco​mmons.​org/​licen​ses/​by/4.​0/. The Creative Commons Public Domain Dedication waiver (http://​creat​iveco​
mmons.​org/​publi​cdoma​in/​zero/1.​0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Jiang et al. BMC Molecular and Cell Biology(2023) 24:15

can provide new ideas and theoretical foundations for theoccurrence, development, and treatment of several disorders [3]. miRNA is a kind of endogenous, short-sequence,
non-coding single-stranded sRNA, generally consistingof 18–25 nucleotides [21]. The mechanism of miRNAaction is mainly divided into the two following categories:
one, whereby miRNA closely binds to the ORF regionof the mRNA, forming a double-stranded structure forits degradation; the other, whereby the miRNA loosely
binds to the 3’UTR region of the mRNA, thus affecting
its post-transcriptional translation [20, 31, 43]. miRNA
mainly acts on the target gene by cutting off the target
gene mRNA molecule, preventing the translation, andbinding to and inhibiting the target gene, thereby playing a role in cellular differentiation, apoptosis, proliferation, metabolism, homeostasis, signaling transduction,
auto-immunity, inflammatory responses, tumor intervention, and various other physiological and pathologicalprocesses [8, 9,16]. miRNA inhibits tumor cell growth,is downregulated in most tumor tissues, and affects
tumor cell proliferation and invasion through the regulation of target proteins [44]. miRNA plays a critical rolein tumor cell metastasis, which provides new ideas forthe diagnosis and treatment of these patients [2, 4]. Specific miRNAs can reflect the tissue origin of tumors andmolecular typing can be conducted to predict the prognosis and estimate the treatment sensitivity of patients,and even directly participate in the treatment [19, 48].
microRNA-125b is differentially expressed across tumor
types and can regulate cell proliferation and apoptosis.
miR-125b-5p shows an inhibitory effect on the invasion and migration of ovarian cancer cells [26, 42]. As apromising new plasma biomarker, hsa-miR-125b-5p canbe employed as one of the biomarkers of colorectal cancer because its expression is closely associated with colorectal cancer [10, 18]. Therefore, the hsa-miR-125b-5p
probe is commonly used for detecting colorectal cancer.
However, in nasopharyngeal carcinoma, the research andapplication of hsa-miR-125b-5p are elusive.
LncRNA is an RNA molecule with a transcript length
exceeding 200 nucleotides [41]. Although lncRNAs
do not encode a protein, they often function as RNA
to regulate protein coding at different levels including transcriptional, post-transcriptional, and epigenetic regulation [5]. LncRNAs can interact with DNA,mRNA, and proteins [39]. These include direct interaction with the promoter region of the coding genes,
mediation of histone modification, chromatin remodeling, regulation of variable mRNA precursor splicing, orformation of endogenous siRNA molecules [6]. Abnormal expression of lncRNAs in various cancer samples
including nasopharyngeal carcinoma is relevant for
regulating the proliferation, invasion, migration, andPage 2 of 12

apoptosis of tumor cells [11, 40]. However, studies on
lncRNAs in nasopharyngeal carcinoma are scarce, andthe existing reports mainly focus on a single lncRNA.
Thereby, an in-depth study of the mechanism oflncRNA action in nasopharyngeal carcinoma is of greatsignificance for its occurrence and development, whichis conducive to early diagnosis, targeted therapy aswell as improved prognosis of these patients. Recently
S-phase cancer-associated transcript 8 (SCAT8), a key
tumor marker of S-phase lncRNA, has gained researchtraction [1]. However, studies focused primarily on themechanism of SCAT8’s action in tumors are lacking.
Scavenger Receptor Class B type I (SR-BI), encodedby Scavenger Receptor Class B Member 1 (SCARB1), is
the plasma membrane receptor of high-density lipoprotein (HDL) cholesterol, mediating the transfer between
cholesterol and HDL [24]. The proliferation and invasion of tumor cells, as well as the enhanced ability toadapt to the tumor microenvironment, are often closely
related to the abnormally active cholesterol metabolism[15, 45]. Therapeutic strategies targeting cholesterol synthesis and its related proteins may shed new insight into
the treatment of tumors [46]. Studies have confirmedthat SR-BI causes proliferation, migration, and enhancement of tumor cell growth in xenograft tumor models ofbreast cancer [17]. In some rare studies on SR-BI in nasopharyngeal carcinoma, increased expression of SR-BI
was found to be related to enhanced cell migration ability [47]. CircRNA-scavenger receptor class B member 1(Circ-SCARB1) is high in 30 pairs of renal cell carcinoma
tissue cell lines [27]. Down-regulated expression of circSCARB1 can restrain cell proliferation, migration, andinvasion, and induce cellular apoptosis [25]. However,studies of SCARB1 in lncRNA are scarce, and the regulatory mechanism underlying its receipt of ncRNA in nasopharyngeal carcinoma remains elusive. Therefore, theregulation of SCARB1 in nasopharyngeal carcinoma wasstudied herein.
In this study, we found that SCAT8 served as a ceRNA
for miR-125b-5p to control SCARB1 expression in nasopharyngeal carcinoma. First, we identified the regulatory functions of SCAT8, miR-125b-5p, and SCARB1 in
the malignant progression of nasopharyngeal carcinoma.
Next, we predicted and verified SCAT8’s binding to miR125b-5p and molecular regulation of SCARB1. Finally, we
found that the SCAT8/miR-125b-5p axis could regulate
the proliferation and migration of nasopharyngeal carcinoma cells through SCARB1. This study lays a theoretical foundation for the development of targeted therapy
and improvement in the prognosis of patients with nasopharyngeal carcinoma.

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Methods

Table 1 The primers in RT-qPCR

Data collection

SCAT8

The mRNA, lncRNA, and miRNA sequencing data of
58 nasopharyngeal carcinoma patients and their corresponding clinical information were downloaded fromThe Cancer Genome Atlas (TCGA) database for cluster
and Cox regression analyses.

Forward:

5’-TGG​TTG​GGC​TTG​AAA​GTG​TA-3’

Reverse:5’-CCA​GCT​GGT​T TT​CAG​TCT​CC-3’

miR-125b-5p Univer- 5’- TCC​C TG​AGA​CCC​TAA​C TT​GTGA-3’
sal R-primer:
SCARB1

Forward:

5’-TGC​AGT​GTT​TCA​CCT​TGC​AT-3’

Reverse:5’-GCC​TCG​GAA​AAC​AAC​T TC​TG-3’

Univariate Cox analysis

Univariate Cox analysis was performed to evaluate theoverall survival rate and status of nasopharyngeal carcinoma patients in TCGA database. mRNA, lncRNA,and miRNA data of nasopharyngeal carcinoma patientswere assessed based on a p-value < 0.05.
Differential gene expression analysis

The data for differential expression analysis was normalized using the limma package. By comparing thedifferentially expressed genes between nasopharyngeal
carcinoma tumor samples and paracancerous samples,
mRNAs, lncRNAs, and miRNAs were analyzed basedon the conditions, logFC ≥ 1 and p-value < 0.05.
Gene enrichment analysis

The biological regulatory effects of differentiallyexpressed genes for mRNAs were evaluated by geneenrichment and pathway analyses. The overlapping
genes enriched in different signaling pathways wereanalyzed using the R package, clusterProfiler, following differential gene expression analysis. If thep-value < 0.05, the enrichment of genes in the corresponding pathways was regarded as significant.

The primer sequences used for RT-qPCR are listed inTable 1.
Dual‑luciferase reporter assay

PGL4.15-control luciferase was inserted into the vector and the wild-type (WT) and mutant (Mut) SCAT8 3’
UTR regions were amplified by PCR. C666-1 cells wereseeded into the 24-well plate. After cell adhesion, WT
and Mut SCAT8 were transfected into C666-1 cells with
Lipofectamine 3000 (Invitrogen). After transfection, cells
were collected and the luciferase activity was detectedon the Dual-Luciferase reporter assay system (Promega).
The relative firefly luciferase activity was normalizedagainst the renilla luciferase activity.
Cell proliferation assay

The CCK-8 reagent was used to detect cell proliferation.
Briefly, C666-1 cells were inoculated into 96-well plates at
a density of 1000 cells per well. After 48 h of cell growth,
10 μL of the CCK8 reagent was added per well. Next, thecells were cultured at 37 ℃. After 3 h, the absorbance values of wells were detected at 450 nm using a microplate
reader.

Cell culture

The nasopharyngeal cancer cell line, C666-1, was purchased from ATCC and grown in RPMI-1640 supplemented with 10% FBS, 100 μg/mL penicillin, and100 μg/mL streptomycin. The cells were maintained ina humidified incubator with 5% ­CO2 at 37 ℃.
Real‑time quantitative
polymerase chain reaction (RT‑qPCR)

Total RNA was isolated from the C666-1 cell line following the kit protocol. Subsequently, the cDNA wassynthesized by reverse transcription. RT-qPCR was
performed following the manufacturer’s instructions. The levels of expression of 5 lncRNAs related
to mRNA were determined by calculating the ratio of
the transcripts in the samples. GAPDH was used as theendogenous control to calculate the relative expression.

Pearson’s correlation analysis

The value of the Pearson correlation coefficient varies from—1 to 1 and it is used to detect the correlation
between two variables. When the Pearson correlation
coefficient is 1, it means that all data points of the variable are on a straight line and the variable Y increases with
the increase in variable X. When the Pearson correlation
coefficient is -1, it implies that all data points fall on astraight line. When the Pearson coefficient is 0, it indicates that the two variables have no linear relationship.
Plotting the receiver operating characteristic curve (ROC)

The area under the curve (AUC) indicated the sensitivity of lncRNA for survival. The diagnostic value of theceRNA axis in nasopharyngeal carcinoma was evaluatedusing the ROC.

Jiang et al. BMC Molecular and Cell Biology(2023) 24:15

Transfection

Inhibitors and mimics of miRNAs were bought from
Biomics (Nantong, China). The ASOs of lncRNA SCAT8
were purchased from RiboBio (Guangzhou, China).
Inhibitors, mimics, and ASOs of miRNA were transfected into C666-1 cells using Lipofectamine 3000 (Invitrogen, US) following the manufacturer’s instructions.
shRNA of SCARB1 was inserted in the PLKO.1 plasmid.
The shRNA sequence was shSCARB1 5’-AUA​AUC​CGA​
ACU​UGU​CCU​UGA​AGG​G-3’.
Transwell migration assay

Cell migration ability was detected by the Transwellmigration assay. The Transwell chamber was placed into
a 24-well plate and the starved cells were resuspended ina serum-free medium. Subsequently, 200 μL cell suspension per well was added into the chamber. The complete
medium was added to the bottom of the chamber. After
12–24 h of cell culture, the inner and outer layers of theculture medium were discarded and cells were washedwith PBS. Polyformaldehyde was used to fix cells. After
15 min of fixation, 0.2% crystal violet dye was used tostain the cells and PBS was used to wash the residual dye.
A cotton swab was used to wipe-off non-migrated cells
and images of migrated cells were captured.
Construction of the lncRNA‑miRNA‑mRNA ceRNA network

The ceRNA network of lncRNA-miRNA-mRNA wasconstructed by predicting the correlations among
lncRNAs, miRNAs, and mRNAs based on the ceRNA
theory. miRNA was a key component of the ceRNA network, playing a central role in connecting lncRNAs andmRNAs. The network was visualized using the Cytoscape
software.
Statistical analysis

The experimental data are presented as mean ± standard
deviation of three independent experiments. LSD-t test
was used for statistical analysis between two groups and
ANOVA was performed for comparison among multiple
groups. When the p-value was less than 0.05, the results
were considered statistically significant. GraphPad Prism
software was used for the analysis and production of statistical charts and the Image J software was used for thegrayscale analysis of protein blots. The Kaplan–Meiermethod was used for plotting the survival curve.

Results
Screening of lncRNAs, microRNAs, and mRNAs related
to the progression of nasopharyngeal carcinoma

During tumor development, the expression of tumor
markers changes significantly with tumorigenesis. Several markers are oncogenes or tumor suppressor genes,

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regulating the malignant progression of tumors. However, the role of the ceRNA axis comprising these genes
in nasopharyngeal carcinoma has not been clarified. To
assess its underlying mechanism, differential expression
analysis, and univariate Cox analysis were performed
based on the mRNAs, lncRNAs, and miRNAs in nasopharyngeal carcinoma tumor samples and paracancerous samples (Fig. 1A, B and Fig. S1A). The upregulated
oncogenes and downregulated suppressor genes in nasopharyngeal carcinoma were extracted after obtaining theoverlapping differentially expressed genes and survivalrelated genes, including 406 protein-coding mRNAs and
165 lncRNAs (Fig. 1C and Fig. S1B).
Enrichment of tumor progression‑related genes
in nasopharyngeal carcinoma

In order to further assess the importance of these genes
in nasopharyngeal carcinoma tumors, GSEA was performed for evaluating mRNA enrichment. The enrichment of the C2 pathway indicated that the genes were
chiefly overrepresented in cell cycle-related pathways(Fig. 2A and Fig. S2A). C5 GO enrichment indicated thatthe genes were also involved in cell cycle, cell proliferation, and cell migration (Fig. 2B and Fig. S2B).
SCAT8/miR‑125b‑5p/SCARB1 may be an important ceRNA
axis regulating nasopharyngeal carcinoma

According to the above results, the ceRNA network
was constructed. It was found to regulate the malignant progression of nasopharyngeal carcinoma basedon the overlapping mRNAs and lncRNAs. The miRNAs
were obtained from differential expression analysis. The
network consisted of five lncRNAs, two miRNAs, six
mRNAs, and 18 groups of interaction axes (Fig. 3A andFig. S3A). Among mRNAs, SCARB1 played a significant
role in regulating the progression of nasopharyngeal carcinoma. SCARB1 is closely related to the proliferationand invasion of tumor cells, which can enhance tumor
adaptability to the microenvironment and inhibit cell
apoptosis. So we focused on the ceRNA axis that regulated SCARB1. The results showed that AC010595.1/
miR-125b-5p, AC025244.1/miR-125b-5p, LINC01179/
miR-125b-5p, LINC02584/miR-125b-5p, and SCAT8/
miR-125b-5p axes likely regulated SCARB1 (Fig. 3B).
SCAT8 may regulate the expression of SCARB1

To further assess the main lncRNA regulating SCARB1,
differential expression analysis and survival analysis were performed for AC010595.1, AC025244.1,
LINC01179, LINC02584, and SCAT8. The results ofdifferential expression analysis suggested that five lncRNAs were significantly overexpressed in nasopharyngeal carcinoma (Fig. 4A). Therefore, these lncRNAs

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Fig. 1 The results of univariate cox and differential expression analysis of mRNAs, lncRNAs and miRNAs from the TCGA-NPC database. Volcano plotsof differential expression analysis of mRNAs, lncRNAs and miRNAs (A). The top ten results of between univariate cox and differential expression
analysis in mRNAs, lncRNAs and miRNAs (B). Heatmaps of mRNAs, lncRNAs and miRNAs (C)could be the reason for the high expression of SCARB1
in nasopharyngeal carcinoma. According to the resultsof survival analysis and correlation analysis, thelncRNA SCAT8 showed a high correlation with worse
survival and the expression SCARB1 (Fig. 4B, Fig. S4AE). Moreover, the AUC curve showed that the survival
sensitivity of SCAT8 was significant (Fig. 4C). To further screen these results, ASO was used to knock down
AC010595.1, AC025244.1, LINC01179, LINC02584,
and SCAT8 (Fig. 4D). By detecting the changes in the
nucleic acid levels of SCARB1, SCAT8 was found to
significantly regulate the expression of SCARB1. However, no statistically significant difference was observed
when other lncRNAs were knocked down. Thus,
SCAT8 may be the main lncRNA regulating SCARB1.

SCAT8, miR‑125b‑5p, and SCARB1 regulate the malignant
progression of nasopharyngeal carcinoma

According to the above predictions, SCAT8/miR125b-5p/SCARB1 may be the ceRNA axis regulating theoccurrence and development of nasopharyngeal carcinoma. However, the effect of this ceRNA axis in themalignant progression of nasopharyngeal carcinoma has
not been clarified. Therefore, nasopharyngeal carcinoma
samples were divided into the SCAT8 high expression
group and SCAT8 low expression group for differential expression analysis and enrichment analysis (miR125b-5p and SCARB1 for the same analysis). The results
indicated that the regulatory genes were mainly enrichedin cell proliferation and cell migration among other pathways (Fig. 5A and Fig. S5A-C). To further verify the effectof the three genes on the proliferation and migration of

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Fig. 2 C5 GO fuctional and C2 pathway enrichment analysis of the diferentally exressed mRNAs. Bubble diagram of C2 pathway analysis of mRNAs(A). Bubble diagram of C5 GO analysis of mRNAs (B)Fig. 3 Construction of nasopharyngeal carcinoma related ceRNA network by integrated analysis. LncRNA-miRNA-mRNA regulatory axes extracted
from this ceRNA network (A). The LncRNA-miRNA-mRNA pattern diagram (B)nasopharyngeal carcinoma cells, CCK8 and Transwellmigration assays were conducted after inhibiting thethree genes. These results were consistent with those ofenrichment analysis (Fig. 5B, C). The above results indicated that the three genes could separately regulate themalignant progression of nasopharyngeal carcinoma.
LncRNA SCAT8 and miR‑125b‑5p can regulate the SCARB1
protein in nasopharyngeal carcinoma

To further assess the relationship among SCAT8, miR125b-5p, and SCARB1, ASO was used to knock down
SCAT8. The changes in the levels of miR-125b-5p and
SCARB1 expression were detected. The results showedthat when SCAT8 was knocked down, the expressionof SCARB1 was downregulated, while the expressionof miR-125b-5p was upregulated. Next, we detected
the changes in SCAT8 and SCARB1-related pathways
after overexpression and inhibition of miR-125b-5p
with corresponding mimic and inhibitor, respectively.
The results showed that SCAT8 and SCARB1-related
pathways were negatively regulated by miR-125b-5p(Fig. 6A). In order to explore whether SCAT8 was
the ceRNA of miR-125b-5p, the predicted bindingsites of SCAT8 and miR-125b-5p were complementarily mutated (Fig. 6B). The reporter gene experiment
was used to detect the effect of miR-125b-5p mutation on luciferase activity. Our results suggested thatthe luciferase activity of wild SCAT8 was regulated
by miR-125b-5p, while that of the mutant SCAT8 was

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Fig. 4 SCAT8 may regulate the mRNA level of SCARB1. The expression of pontential lncRNAs between tumor and normal tissues in nasopharyngeal
carcinoma patients. The red box indicates the tumor tissue and the black box indicates the normal tissue (A). The survival curves of pontential
lncRNAs in nasopharyngeal carcinoma patients. Black lines indicate low expression of lncRNA and red lines indicate high expression of lncRNA (B).
The ROC curves of the 5 lncRNAs in nasopharyngeal carcinoma (C). SCARB1 mRNA levels in C666-1 cells with 5 potential lncRNAs knockdown by
ASO (D)not (Fig. 6C). All the above results indicated that thelncRNA SCAT8 and the microRNA miR-125b-5p could
regulate the expression of the mRNA SCARB1 in nasopharyngeal carcinoma. These results were consistent
with the trend of the ceRNA axis.

As the ceRNA of miR‑125b‑5p, SCAT8 regulates SCARB1
to affect the malignant progression of nasopharyngeal
carcinoma

Next, we predicted the enrichment of two or more genes
regulated by the SCAT8/miR-125b-5p/SCARB1 axis in
the malignant progression of nasopharyngeal carcinoma.

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Fig. 5 SCAT8, miR-125b-5p and SCARB1 proteins regulate nasopharyngeal carcinoma malignant progression. The C5 GO functional enrichment
analysis of the related biological processes of differential expression genes in low expression group and high expression group among SCARB1,
SCAT8 and miR-125b-5p (A). The CCK8 results in C666-1 cells with SCARB1, SCAT8 and miR-125b-5p knockdown or inhibited (n = 3) (B). The
Transwell results in C666-1 cells with SCARB1, SCAT8 and miR-125b-5p knockdown or inhibited (C)The gene enrichment results showed that the proliferation and migration pathways were particularly significantly overrepresented (Fig. 7A, B and Fig. S6A-D). To
verify this finding, SCAT8 was knocked down in thenasopharyngeal carcinoma cell line, C666-1, in control
and experimental groups. The experimental group wastreated with a miR-125b-5p inhibitor. The proliferationand migration of C666-1 cells were detected by CCK-8
and Transwell migration assays, respectively. The results
showed that knocking down SCAT8 inhibited the proliferation and migration of nasopharyngeal carcinoma
cells in the control group but this result was no longer

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Fig. 6 SCAT8 regulates SCARB1 protein and affects the malignant progression of nasopharyngeal carcinoma tumors. RT-qPCR assays were used
to detect the mRNA levels of miR-125b-5p, SCARB1 and SCAT8 in C666-1 cell treated with ASO, mimic or inhibitor (n = 3) (A). Predicted bindingsite between miR-125b-5p and SCAT8 (B). Luciferase assays were performed to test the effect of miR-125b-5p on wild-type or mutant SCAT8 after
treating with miR-125b-5p mimic (n = 3) (C)observed after using the mimic (Fig. 7C, D). The above
results indicated that as the ceRNA of miR-125b-5p,
SCAT8 regulated SCARB1 and affected the malignant
progression of nasopharyngeal carcinoma.

Discussion
Nasopharyngeal carcinoma is a highly malignant tumorof the nasopharynx. Due to the obvious early common symptoms, the mortality rate of nasopharyngeal
carcinoma remains high. Patients often undergo medical examinations only after they present with obvious symptoms. By this time, more than 70% of patients
have progressed to the advanced stage of the disease.
Nasopharyngeal carcinoma has the characteristics ofhigh invasiveness, rapid metastasis, poor prognosis, andcomplex complications [35]. The traditional treatmentof nasopharyngeal carcinoma includes surgery, chemotherapy, and radiotherapy but given the deep anatomicalsite and complex anatomical structure, the curative effectof surgery is limited. Radiotherapy and chemotherapy

often lead to decreased immunity and the occurrence of
secondary tumors. Therefore, it is significant to develop
new and effective treatment methods, enhance the treatment rate and prognosis of patients with nasopharyngeal
carcinoma, and effectively reduce their complications.
LncRNA is a kind of ncRNA, which exert diverse biological functions. LncRNAs may participate in the occurrence and development of nasopharyngeal carcinoma
by interacting with adjacent coding transcripts or proteins. LncRNA can serve as ceRNA and miRNA, thereby
regulating each other, participating in the expression oftarget genes, and playing a key role in tumor processes.
Both lncRNAs and miRNAs are involved in the inflammatory cascade of scavenger receptors and subsequent
regulation of the lipid metabolism pathways [36]. The
expression of SCARB1 in gastric cancer tissues is significantly down-regulated, and its mechanism of action
may be associated with the post-transcriptional regulation of SCARB1 mRNA by miRNA [7, 29]. For example,
during tumor occurrence, high levels of miR-199 can

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Fig. 7 SCAT8 regulates SCARB1 by miR-125b-5p to affect the malignant progression of nasopharyngeal carcinoma. Venn diagrams represent theoverlapped genes in low expression group and high expression group among SCAT8, miR-125b-5p, and SCARB1 (A). The bubble diagram of GO
enrichment analysis of overlapped genes by DAVID database (B). The CCK8 results of C666-1 cells with knockdown SCAT8 treated with NC mimic or
miR-125b-5p mimic (n = 3) (C). The Transwell results of C666-1 cells with knockdown SCAT8 treated with NC mimic or miR-125b-5p mimic (D)bind to the 3’ UTR of the SCARB1 mRNA, thus inhibiting the expression of SCARB1 [13, 38, 45, 46]. Reduced
expression of SCARB1 can induce gastric cancer, which
may be related to the regulation of the immune function of CDB + 7 lymphocytes through its expression in
the tumor microenvironment. However, the ceRNA network of SCARB1 has not been reported in nasopharyngeal carcinoma. Herein, we found that SCAT8 is a ceRNAof miR-125b-5p that governs the expression of SCARB1
in nasopharyngeal carcinoma. The establishment of aco-regulatory network of SCARB1 and ncRNA in nasopharyngeal carcinoma provides a potential theoreticalbasis for the diagnosis and treatment of these patients.

SR-BI encoded by the SCARB1 gene acts as a multifunctional HDL receptor that promotes the uptakeof cholesteryl esters by the liver. Recent studies haveshown that HDL has excellent properties as an anticancer drug carrier, and the SCARB1 gene has entered thefield of anticancer research [23, 47]. SR-BI is thought tobe critical for cholesterol and lipoprotein metabolism inbreast and prostate cancer [22, 48]. In all nasopharyngeal carcinoma cell lines and 75% of nasopharyngeal carcinoma biopsies, SR-B1 is overexpressed. Thus, SR-B1
is a potential biomarker for nasopharyngeal carcinoma[38]. The results of the enrichment analysis showed that
SCAT8, miR-125b-5p, and SCARB1 could all be involved

Jiang et al. BMC Molecular and Cell Biology(2023) 24:15

in cellular proliferation and migration-related pathways,
and their regulation by the SCAT8/miR-125b-5p axis wasdependent on SCARB1 to a certain extent. Therefore,
the SCAT8/ miR-125b-5p/SCARB1 axis may regulate
the malignant progression of nasopharyngeal carcinoma
through SCARB1.
In conclusion, the regulation of SCARB1 was by modulating the levels of SCAT8 and miR-125b-5p expression.
Mechanistically, SCAT8 is a molecular sponge of miR125b-5p which competitively regulate the expressionof an important oncogene, SCARB1, in nasopharyngeal
carcinoma. Taken together, the SCAT8/ miR-125b-5p/
SCARB1 axis may regulate the malignant progression ofnasopharyngeal carcinoma through SCARB1.

Supplementary Information
The online version contains supplementary material available at https://​doi.​
org/​10.​1186/​s12860-​023-​00477-2.
Additional file 1: Fig. S1. The results of univariate cox and differential expression analysis of mRNAs, lncRNAs and miRNAs. PCA diagrams of
mRNAs,lncRNAs and miRNAs (A). Venn diagram displaying the intersection genes of the COX analysis and the DEG analysis of mRNAs andlncRNAs (B). Fig. S2. C5 GO fuctional and C2 pathway enrichment analysisof the differentally exressed mRNAs. C2 pathway enrichment analysis ofregulated mRNAs (A). C5 GO enrichment analysis of regulated mRNAs(B). Fig. S3. Construction of ceRNA network by integrated analysis(A). Fig.
S4. SCAT8 may regulate the mRNA level of SCARB1. Pearson’s correlation coefficient between SCARB1 and 5 lncRNAs (AC010595.1,
AC025244.1,LINC01179, LINC02584 and SCAT8) (A-E). Fig. S5. The differential expression analysis of the SCAT8,miR-125b-5p and SCARB1 in nasopharyngeal carcinoma patients. The volcano plotsand the GSEA of theDEGs in low expression group and high expression groupamong SCARB1(A), SCAT8 (B) and miR-125b-5p (C). Fig. S6. SCAT8 regulates SCARB1
by miR-125b-5p to affect themalignant progression of nasopharyngeal
carcinoma. The bubble diagram of GO enrichment analysis of SCAT8 and
SCARB1 overlapped genes by DAVID database (A).The bubble diagram of
GO enrichment analysis of miR-125b-5p and SCARB1overlapped genes by
DAVID database (B). Venn diagrams represent the overlappedgenes in low
expression group and high expression group between SCAT8 and SCARB1(C). Venn diagrams represent the overlapped genes in low expressiongroup and high expression group between miR-125b-5p and SCARB1 (D).
Authors’ contributions
CJ conceived and designed the experiments, analyzed parts of bioinformatics
data and wrote the manuscrip. YZ carried out the experiments. DF, KY and XH
analyzed the data, prepared fgures and tables. QX guided the whole researchand corrected the manuscript. All authors reviewed the final manuscript. The
author(s) read and approved the final manuscript.
Funding
All authors declare that no funds, grants or other supports were received during the preparation of this manuscript.
Availability of data and materials
Bioinformatics data in this manuscript was obtained from the database of
TCGA [https://​portal.​gdc.​cancer.​gov/].

Declarations
Ethics approval and consent to participate
This research does not contain any studies with human participants oranimals.

Page 11 of 12

Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Received: 20 January 2023 Accepted: 20 March 2023

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