Bioinformatics analysis of IGFBP-3 and research advances in brain tumors Running title：Protein structure and biological function

doi:10.21203/rs.3.rs-2126886/v1

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Bioinformatics analysis of IGFBP-3 and research advances in brain tumors Running title：Protein structure and biological function

https://doi.org/10.21203/rs.3.rs-2126886/v1

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OBJECTIVE：Bioinformatics analysis of human insulin-like growth factor-binding protein-3 (IGFBP-3) molecules and genes, and review of the pathways and regulatory mechanisms of IGFBP-3 in brain tumors ( especially gliomas ) .

METHODS：Analysis of IGFBP-3 protein open reading frame（ORF）and physicochemical properties, secondary structure, tertiary structure, signal peptide and nuclear localization signal, transmembrane structure, domain, phosphorylation and glycosylation of IGFBP-3 protein using molecular biology tools Sites, proteins and biological functions that interact with IGFBP-3 proteins.

RESULTS：The molecular formula of IGFBP-3 protein is C1390H2230N418O422S21. 104 ORFs were found in the mRNA sequence of IGFBP-3. The longest ORF is ORF2, which is 519 bp in total and can encode 172 amino acids.It is an unstable hydrophilic secreted protein, the secondary structure is mainly ring structure and helical structure, containing 67 phosphorylation sites and 3 N-glycosylation sites, no transmembrane region, 3 main domains, Having nuclear localization signals exerts biological effects in the nucleus. IGFBP-3 can participate in biological functions such as apoptosis, negative regulation of cell proliferation, negative regulation of signal transduction, and regulation of insulin-like growth factor receptor signaling pathway.

CONCLUSIONS：The predicted results provide a basis for further research on the structure and biological function of IGFBP-3 gene and its encoded protein.

Insulin-like growth factor-binding protein-3

bioinformatics

brain tumors

tumor microenvironment

Regulatory mechanisms

Insulin-like growth factor binding proteins (IGFBPs) are a family of proteins that can specifically bind to IGF-1, regulate IGF activity and have tissue specificity, of which IGFBP-3 is the most abundant in serum.IGFBP-3 has attracted the most attention in the past few years for its role in tumorigenesis and development. Studies have found that it can regulate IGF-1 by prolonging the half-life of IGF-1 and limiting its binding to IGF-1R Biological activity, can also participate in multiple cell signaling pathways (such as Smad/TGF-b pathway) to regulate the occurrence, development, metastasis and recurrence of tumors in the case of IGF-1-independent;It can also be used as a negative regulator to inhibit tumor cell proliferation, invasion and migration, and predict the risk of tumor occurrence^[1].IGFBP-3 has been widely reported in lung cancer^[2], colon cancer^[3], breast cancer^[4]and prostate cancer^[5], but so far there are few reports on IGFBP-3 bioinformatics analysis and related studies in brain tumors.In this study, the gene sequence, physicochemical properties, secondary structure, tertiary structure, phosphorylation and glycosylation sites, signal peptides, subcellular localization, transmembrane regions, domains and interacting proteins of IGFBP-3 were bioinformatics, In order to further explore the biological function and molecular mechanism of human IGFBP-3, and to review the action pathway and regulatory mechanism of IGFBP-3 in brain tumors (especially glioma).

1.1 Materials

The mRNA sequence of the human IGFBP-3 gene（GI：224809341）and its encoded protein amino acid sequence（NP_001013416. 1）were obtained from the National Center for Biological Information (NCBI)（https://www.ncbi.nlm.nih.gov）.

1.2 Methods

Analysis and definition (minimum 30 bases) of the ORF of the IGFBP-3 gene using the open reading frame (ORF) Finder online tool(https://www.ncbi.nlm.nih.gov/orffinder).The physicochemical properties of IGFBP-3 protein were analyzed by ProtParam of ExPASy database（https://web.expasy.org/protparam）.The secondary structure of IGFBP-3 protein was analyzed by Predict-Protein（https://predictprotein.org）,SOPMA（https://npsa-prabi.ibcp.fr/cgi-bin/secpred_sopma.pl）analysis of IGFBP-3 protein secondary structure composition.The three-dimensional structural model of IGFBP-3 protein was constructed by SWISS-MODEL online tool（https://swissmodel.expasy.org/interactive）.The transmembrane region of IGFBP-3 protein was predicted by TMHMM online tool（http://www.cbs.dtu.dk/services/TMHMM）.The phosphorylation sites and N-glycosylation sites of IGFBP-3 protein were predicted by NetPhos 3.1（https://services.healthtech.dtu.dk/service.php?NetPhos-3.1）and NetNGlyc 1.0 （https://services.healthtech.dtu.dk/service.php?NetNGlyc-1.0）online tools, respectively.Prediction of the signal peptide of IGFBP-3 protein by neural network model in SignalP4.1（http://www.cbs.dtu.dk/services/SignalP-4.1） of CBS database.The nuclear localization signal of IGFBP-3 protein was predicted by NLStradamus（http://www.moseslab.csb.utoronto.ca/NLStradamus）.The String database（https://string-db.org） was used to predict proteins that interact closely with IGFBP-3 proteins, and the Gene Ontology Consortium（http://geneontology.org） predicted the biological functions of IGFBP-3.

2.1 ORF analysis of IGFBP-3 gene

Analysis and definition (minimum 30 bases) of the ORF of the IGFBP-3 gene using the ORF Finder online. The results show that the nucleotide sequence of the IGFBP-3 gene contains 104 ORFs, the longest ORF is ORF2, a total of 519 bp, which can encode 172 amino acids (Figure 1, Figure 2).

2.2 Physicochemical properties of IGFBP-3 protein

The physicochemical properties of IGFBP-3 protein were predicted by Prot-Param tool, including molecular weight, amino acid composition, fat index, instability coefficient, hydrophilicity, etc.The results showed that the molecular formula of IGFBP-3 is C₁₃₉₀H₂₂₃₀N₄₁₈O₄₂₂S₂₁, the molecular weight is 32222.80 Da, and the protein contains 20 amino acids (Table 1).The number of amino acids is 297. The theoretical isoelectric point is 8.96.The instability index is 51.58, which belongs to alkaline unstable protein. The fat index was 65.08 and the hydrophilicity was -0.547. Therefore, IGFBP-3 is a hydrophilic protein.

Table 1 Amino acid composition of IGFBP-3 protein

amino acid	amount	Proportion%	amino acid	amount	Proportion%
Ala (A)	26	8.8%	Leu(L)	25	8.4%
Arg (R)	23	7.7%	Lys (K)	19	6.4%
Asn (N)	6	2.0%	Met (M)	3	1.0%
Asp (D)	12	4.0%	Phe(F)	5	1.7%
Cys (C)	18	6.1%	Pro (P)	29	9.8%
Gln(Q)	11	3.7%	Ser (S)	27	9.1%
Glu (E)	18	6.1%	Thr(T)	11	3.7%
Gly(G)	24	8.1%	Trp(W)	2	0.7%
His(H)	7	2.4%	Tyr(Y)	9	3.0%
Ile (I)	6	2.0%	Val (V)	16	5.4%

2.3 Secondary structure of IGFBP-3 protein

The secondary structure of IGFBP-3 protein was analyzed by the PROFSec method (neural network algorithm) in the Predict-Protein tool (Figure 3)；Secondary structure composition of IGFBP3 protein analyzed by SOPMA (Figure 4).The results showed that IGFBP-3 protein was mainly circular structure, followed by helical structure.The solvent accessibility analysis was mainly exposed; in the secondary structure, α-helix accounted for 21.55%, random coil accounted for 64.31%, β-turn accounted for 5.05%, and extended chain accounted for 9.09%.

2.4 Tertiary junction of IGFBP-3 protein

The three-dimensional structural model of IGFBP-3 protein was constructed using the SWISS-MODEL online tool.The results show that the model quality evaluation parameter GMQE(GMQE score is a number between 0 and 1, the closer to 1, the higher the model reliability) is 0. 48.QMEAN is 0.57(The QMEAN score near 0 indicates good agreement between the model structure and experimental results of similar size. A score of -4.0 or below indicates a low quality model).The modeling results showed that the amino acid sequence of IGFBP-3 had 100% similarity with its template 7wrq. 1.B, and the three-dimensional structural model of IGFBP-3 protein was better (Figure 5).

2.5 Phosphorylation and glycosylation sites of IGFBP-3

According to NetPhos 3.1 prediction, IGFBP-3 protein contains 67 phosphorylation sites, including 44 serine sites, 10 threonine sites and 13 tyrosine sites (Figure 6).As predicted by NetNGlyc 1.0, IGFBP-3 protein contains 3 N-glycosylation sites, which are located at positions 116, 142 and 205, respectively (Figure 7).

2.6 Signal peptide prediction and nuclear localization of IGFBP-3 protein

The signal peptide of IGFBP-3 protein was predicted by neural network model in SignalP4.1. The results show that: IGFBP-3 is a secreted protein with a signal peptide located at amino acids 1-27 (Figure 8).Prediction of nuclear localization Signal (NLS) of IGFBP-3 by NLStradamus.The NLS signal peptide mediates the entry of proteins into the nucleus through the nuclear pore complex.The NL Stradamus prediction results in this study showed that there was NLS in the amino acid sequence of IGFBP-3, suggesting that IGFBP-3 could be located in the nucleus (Figure 9).

2.7 Transmembrane region prediction of IGFBP-3 protein

The transmembrane region of IGFBP-3 protein was predicted by TMHMM online tool.The results show that the expected value of amino acids in the transmembrane region of IGFBP-3 protein is 4.23698 (when the value is greater than 18, it means that there is a transmembrane region), so IGFBP-3 protein has no transmembrane region (Figure 10).

2.8 Domain prediction of IGFBP-3 protein

The domain of IGFBP-3 protein was predicted by SMART database.The results showed that IGFBP-3 protein had five conserved regions, including one in the insulin growth factor binding protein homolog (IB) domain, one in the thyroglobulin type I repeat (TY) domain, and the remaining three low-degree complex conserved regions(Figure 11).

2.9 Protein closely interacting with IGFBP-3 and prediction of biological function

The String database was used to predict the proteins closely interacting with IGFBP-3 protein.The results showed that 10 proteins were found that interacting closely with the IGFBP-3 protein (Figure 12).Gene Ontology Consortium analysis of biological functions of IGFBP-3 showed that the types of molecular mechanisms include fibronectin binding, insulin-like growth factor binding, heavy metal ion binding, protein binding, and protein tyrosine phosphokinase activator activity;The physiological processes involved include apoptosis, negative regulation of cell proliferation, negative regulation of protein phosphorylation, negative regulation of signal transduction, positive regulation of apoptosis process, positive regulation of MAPK cascade, cell growth regulation, insulin-like growth factor receptor Signal pathway regulation;Its cellular components include extracellular body, extracellular region, extracellular space, insulin-like growth factor binding protein complex, insulin-like growth factor ternary complex, and nucleus.

3.1 Structure and biological effects of IGFBP-3

IGFBP-3 is the most abundant IGFBP circulating in adult mammals and binds IGF-1 and IGF-2 in the cellular environment.The structure of IGFBP-3 mainly consists of five domains, the most clearly studied are the insulin growth factor binding protein homolog (IB) domain and the thyroglobulin type I repeat (TY) domain.Within each major region there are other corresponding subregions and functional motifs.In the C-terminal region of IGFBP-3, there are functional sites for binding to IGF, heparin, the acid-labile subunit, integrins, and the nucleus^[6].Mutations of three important amino acids (I56, L80 and L81) in the N-terminal IGF domain of IGFBP-3 to glycine (G56, G80 and G81) have been confirmed to affect IGF binding to IGFBP-3^[7].There is also an IGFBP-3 nuclear localization sequence in the C-terminal domain of IGFBP-3.The mutation of leucine at positions 224 and 227 of IGFBP-3 nuclear localization sequence to alanine will affect the binding of IGFBP-3 complex to nuclear receptor^[8];There is a metal-binding domain in the C-terminal domain of IGFBP-3. IGFBP-3 not only binds metals through this site, but also has other intrinsic roles, including the possible binding of IGFBP-3 to integrins through this domain.It is because of the binding sites of these different substances that IGFBP-3 has its biological function diversity.

3.2 The role of IGFBP-3 in brain tumors

The role of IGFBP-3 in tumors is also divided into two mechanisms: IGF-dependent and IGF-independent, and under different mechanisms,IGFBP-3 plays opposite effects of promoting or suppressing cancer in different tumors.The generation of this bidirectional effect is mainly related to the activation of different signaling pathways. As a tumor suppressor, IGFBP-3 inhibits tumor cell adhesion in tumors^[9];Inhibition of epithelial ovarian cancer tumor angiogenesis, and then inhibit tumor metastasis^[10];It affects prostate cancer metastasis^[11];Reduced angiogenesis in non-small cell lung cancer and head and neck squamous cell carcinoma^[12].In terms of the effect of promoting tumor proliferation, IGFBP-3 has antioxidant activity, which can promote epithelial-mesenchymal transition and promote the occurrence and development of tumors.Current studies have confirmed that IGFBP-3 has a significant pro-tumor proliferative effect on lung cancer^[2], triple-negative breast cancer^[13], esophageal squamous cell carcinoma^[14]and other cancers, and IGFBP-3 also has pro-apoptotic or pro-proliferative effects in gliomas.

3.2.1 Antitumor effect of IGFBP-3

IGFBP-3 has tumor suppressor properties. Study shows IGFBP-3 is more strongly associated with glioblastoma than other brain tumors^[15].Yang et al^[16]found that high expression of IGFBP-3 can inhibit the cell proliferation of GBM under certain circumstances,Through experiments on U87MG, they found that miR-21 would target the inhibition of IGFBP-3 gene expression,At the same time, after knockdown of miR-21 in U87MG cells, the expression of IGFBP-3 was increased, and the tumorigenicity of U87 cells was inhibited, indicating that IGFBP-3 can act as a tumor suppressor gene to play a pro-apoptotic effect in GBM.

3.2.2 Cancer promoting effect of IGFBP-3

On the other hand, IGFBP-3 has the effect of promoting tumor proliferation. The study found that IGFBP-2, IGFBP-3 and IGFBP-5 are expressed in GBM and are closely related to the poor prognosis of GBM^[15].For IGFBP-3, Chen et al^[17] found that the expression of IGFBP-3 in glioma tumor center was higher than that in tumor margin and diffuse astrocytoma, which indicated that high IGFBP-3 expression was positively correlated with glioma grade;Moreover, patients with high expression of IGFBP-3 had a shorter overall survival period; IGFBP-3 knockout inhibited cell proliferation of glioma cells, induced cell cycle G2/M arrest and apoptosis.Similarly, Xue et al^[18]showed that the expression of IGFBP-3 in glioma increased with the increase of malignant grade;In vitro studies showed that IGFBP-3 cytokines could promote the proliferation of glioblastoma cell line U87MG, and the proliferation was more obvious with the increase of the dose, indicating that IGFBP-3 played a cancer-promoting role in the evolution of glioma.

3.2.3 The pathway of action of IGFBP-3 in brain tumor

The IGF-dependent pathways for IGFBP-3:The binding of IGFBP-3 to IGF leads to the opening of the corresponding signaling pathway, which has been confirmed to be involved in the progression of glioma^[19],Elevated serum IGF-1 is regarded as one of the recognized diagnostic markers of different tumors, especially brain malignant tumors.Ye Yun et al^[20]detected the changes of IGF-1 and IGFBP-3 in normal brain tissue and glioma tissue by immunohistochemical staining,and were positively correlated with the pathological grade of glioma, suggesting that IGFBP-3 may be Enhanced the pro-tumor proliferative effect of IGF-1.

The IGF-Independent Pathway of IGFBP-3:It was confirmed that the binding ability of IGFBP-3 to IGF-2 and IGF-2 was completely lost when the N-terminal domain I56, L80 and L81 of IGFBP-3 were mutated to glycine^[7].To explore the IGF-independent pathway against this mutant, it has been confirmed that the IGFBP-3 mutant does not cause growth retardation in mice in vivo^[21];The effect of mutant IGFBP-3 is compensated by other related mechanisms, namely the IGF-independent pathway, and this IGF-independent mode of action also contributes to the induction of related gene transcription, thus exerting its effect.Studies have found that the increased expression of IGFBP-3 in drug-resistant non-small cell lung cancer cells promotes the enhanced activity of IGF-1R and the subsequent phosphorylation of protein kinases, suggesting that there may be alternative signaling pathways other than the IGF-1R pathway^[22].Hu YH et al^[23]found that IGFBP-3 regulates the expression and activation of ERK1/2 and affects the expression of the downstream regulatory gene bcl-xL, thereby regulating the apoptosis and survival of U87MG cells.Yang J et al^[24]found that IGFBP-3 can promote the proliferation, colony formation and G1/S phase transition of U87MG and U251 MG cells, which may be related to the activation of ERK signaling pathway and the up-regulation of cyclinE and CDK2 proteins.

3.3 The relationship between IGFBP-3 and the brain tumor microenvironment

At present, it is known that the microenvironment of brain tumors is mainly composed of immune cells, hypoxia microenvironment and various cytokines,etc;The connection between it and IGFBP-3 is introduced as follows:

IGFBP-3 and Immune Cells:In the glioma microenvironment, immune cells activate corresponding signaling pathways to stimulate the expression of certain cytokines^[25].Scully et al^[26]found that high expression of IGFBP-3 in tumor cell interstitium was positively correlated with tumor proliferation under high-fat environment;At the same time, increased CD8 was detected by immunohistochemistry, indicating that knockdown of IGFBP-3 caused CD8+T cell aggregation, which disappeared when knockdown of IGFBP-3 mice were fed a high-fat diet.This indicates that the presence of IGFBP-3 and high-fat environment will limit the anti-tumor proliferation effect of CD8+T and other immune cells.Wolf et al^[27]found that the impaired immune function caused by severe burns leads to a weakened Th1 cell response and an increased Th2 cell response;Although there are still limited reports on IGFBP-3 and immune cells in brain tumors, this brings some enlightenment to study the relationship between IGFBP-3 and immune cells in brain tumors.

IGFBP-3 and Hypoxic Microenvironment:As a significant feature of gliomas, hypoxic microenvironment exists in various processes of glioma development. By interfering with the hypoxic microenvironment of HIF and other molecules, it inhibits the occurrence and development of gliomas^[28].Ragel et al^[29]compared the gene expression profile of U251 glioma cell line under normoxia and hypoxia, and found that hypoxia stress induced the upregulation of IGFBP-3 gene, indicating that hypoxia in glioma could have a corresponding influence on IGFBP-3 gene expression.

IGFBP-3 and cytokines:Cytokines, as important communication media between cells and cells and between cells and tissues in the glioma microenvironment, are closely related to immune cell activation and glioma progression.Karen et al^[30] showed that IGFBP-3 is involved in activating the TGF-β/Smad signaling pathway to promote the proliferation of human cytotrophoblasts.Kjellman et al^[31]confirmed that TGF-β is overexpressed in gliomas and positively correlated with tumor grade.Yang et al^[2]demonstrated that IGFBP-3 promotes TGF-β-mediated epithelial-mesenchymal transition in lung-brain metastatic cancer A549 cells. The above studies thus speculate that IGFBP-3 may also be involved in activating the TGF-β/Smad signaling pathway in gliomas, and directly affect the occurrence and development of gliomas.There are no reports that IGFBP-3 interacts with certain inflammatory factors and exerts effects in brain tumors, and more related studies are expected in the future.

IGFBP-3 binds to receptors in brain tumors and exerts effects

3.4 IGFBP-3 and Membrane Receptors:IGFBP-3 binds to the relevant receptors on the cell membrane surface and exerts corresponding effects through IGF-independent pathways. These receptors include epidermal growth factor receptor, integrin and TGF-β receptors, among others.For the cell membrane surface EGFR, Martin et al^[32]studied the mechanism by which IGFBP-3 enhances EGFR signaling in breast epithelial cells, and the results showed that IGFBP-3 enhanced the activation of EGFR stimulated by EGF and DNA synthesis.Park et al^[33]studied the role of third-generation EGFR-TKIs in drug-resistant lung cancer cells and found that activation of the IGF-1R pathway associated with IGFBP-3 deletion can induce acquired resistance to mutant-selective EGFR-TKIs. Therefore, The combination of IGFBP-3-deficient-related IGF-1R inhibitors and EGFR-TKI inhibitors may be a therapeutic strategy for acquired drug-resistant lung cancer.Integrin β1 silencing can affect exogenous IGFBP-3-mediated tumor cell migration and ERK phosphorylation, suggesting that integrin β1 plays a critical role in the enhancement of IGFBP-3 function^[34].IGFBP-3 can activate and bind to the TGF-β/Smad signaling pathway and related receptors.Schedlich et al^[35]found the interaction between endogenous IGFBP-3 and TGF-β receptor and IGF-1R signaling pathway in osteosarcoma cells. IGFBP-3 attenuates the activation of ERK1/2 and Akt by TGF-β1, and inhibits the induction of TGF-β1. the cell cycle progression and proliferation.

IGFBP-3 and Nuclear receptors:IGFBP-3 has functions of cellular uptake and nuclear uptake, but little is known about how it functions in the nucleus.It has been reported that extracellular IGFBP-3 enters the nucleus via multiple endocytic mechanisms. Its nuclear entry appears to be a common pathway with IGFBP-5, both of which are mediated by binding to nuclear transporter-β.IGFBP-3 can also form a complex with EGFR and interact with DNA-dependent protein kinase to form a nuclear complex after entering the nucleus^[36].There is clearly much more to be researched regarding the nuclear role of IGFBP-3, and since IGFBP-3 was first identified as having nuclear translocation, there has been significant progress in uncovering the role of this translocation and the nuclear function involved in IGFBP-3 in brain tumors. The effect is not fully understood yet.

It has been reported that IGFBP-3 binds to the nuclear receptors retinoic acid X receptor α, Nur77, etc to play a biological role after entering the nucleus of tumor cells.Many studies have shown that retinoic acid X receptor α and IGFBP-3 can bind to each other in the nucleus, which is one of the important proteins that interact with IGFBP-3 in the nucleus^[37].RXRα is a member of the nuclear receptor family, and in the luciferase reporter gene assay, it was confirmed that IGFBP-3 enhances the transcriptional activity of the RXRα ligand SR11235 and the signaling of the RXRα homodimer,Also the interaction of IGFBP-3 and RXRα may involve some residues in the carboxy-terminal domain of IGFBP-3 and residues Thr58 and Arg60 in the amino-terminal domain of IGFBP-3.

After IGFBP-3 forms a complex in the nucleus, it will produce a secretion effect, and then the IGFBP-3 complex will be secreted to the outside of the cell through a nuclear export pathway. On the one hand, it activates organelles such as mitochondria and endoplasmic reticulum; on the other hand, it is secreted to the Extracellular circulation acts on the corresponding parts.Our team previously found that serum and cerebrospinal fluid IGFBP-3 were elevated in children with medulloblastoma, and immunohistochemical analysis showed that MB cells had high expression of IGFBP-3, which was associated with poor prognosis.Few studies have focused on the specific role of IGFBP-3 secretion in brain tumors, but serological detection of IGF-1 and IGFBP-3 may serve as an important screening indicator for biochemical detection of gliomas in the future.

In this study, various bioinformatics analysis tools were used to analyze the molecular structure and biological function of IGFBP-3 protein.ORF Finder found 104 ORFs in the IGFBP-3 mRNA sequence, and the longest ORF was ORF2, with a total of 519 bp and encoding 172 amino acids.The molecular formula of IGFBP-3 protein is C₁₃₉₀H₂₂₃₀N₄₁₈O₄₂₂S₂₁.The instability index was 51.58, and the hydrophilicity was -0.547, indicating that it was an unstable hydrophilic protein.The secondary structure of IGFBP-3 protein is mainly ring structure and helical structure. IGFBP-3 protein has no transmembrane region and can function in the cytoplasm and nucleus.Using the SMART database, it was predicted that IGFBP-3 protein contains 5 conserved regions, including 1 insulin growth factor binding protein homolog (IB) domain, 1 located in thyroglobulin type I repeat (TY) domain, and the remaining 3 a low-complexity conservative region.IGFBP-3 protein contains 67 phosphorylation sites, which are 44 serine sites, 10 threonine sites and 13 tyrosine sites; 3 N-glycosylation sites,The mechanism of action of IGFBP-3 protein is still unclear.In this study, the systematic bioinformatics analysis of IGFBP-3 protein has laid the foundation for further research on the function and mechanism of IGFBP-3 protein. At the same time, it was found that IGFBP-3 protein is highly correlated with MMP2.And summarizes the research progress of IGFBP-3 in brain tumors (especially gliomas), which provides a theoretical basis for the next step to study the mechanism of IGFBP-3.

Current studies have shown that IGFBP-3 is closely related to the development and prognosis of various tumors, especially the current research on glioma, which is a research hotspot.As a key site in multiple signaling pathways, IGFBP-3 can bind to receptors at the corresponding positions to form complexes, and plays an important role in the occurrence, development, invasion and metastasis of gliomas.The study of the structure and function of IGFBP-3 may provide new ideas for human glioma research, and provide new targets for the gene therapy of glioma, and become a new approach for glioma treatment.

Funding：This work was supported by Natural Science Foundation of Xinjiang Uygur Autonomous Region (Grant numbers：2022D01C257).

Conflict of interest：The authors have no fnancial confict of interest.

Author contributions:FGF and LW Study design；YRZT,LYB,MLH and LD Software application analysis；FGF wrote the manuscript; WYX analyzed data and revised the manuscript;All authors read and approved the fnal manuscript.

Data availability：The data supporting the findings of this study are available online.

Ethics approval：No ethical support is required for this study.

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Bioinformatics analysis of IGFBP-3 and research advances in brain tumors Running title：Protein structure and biological function

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