Abstract
Glioma is a common central nervous system tumors in children. CMYC has a range of functions that are disrupted in various tumor cells, and may contribute to the occurrence and development of glioma. Two CMYC single nucleotide polymorphisms (rs4645943C>T and rs2070583 A>G) were genotyped in 190 cases and 248 controls from Wenzhou and Guangzhou hospitals. After adjusting for age and sex, odds ratio and 95% confidence interval values were calculated by logistic regression to evaluate the correlation between CMYC gene polymorphisms and glioma risk; no significant associations were detected. These results require future validation in a larger sample cohort.
Introduction
Glioma is an intracranial tumor that originates from glial cells, and one of the most common malignant tumors in children, accounting for 45%–55% of pediatric intracranial tumors (Citation1,Citation2). The overall annual incidence of glioma is 3–8 per 100000, and varies according to glioma subtype, among which astrocytoma is the most common, accounting for 17.4%, with ependymoma accounting for 10.2%, and oligodendroglioma for 5.8%. Treatment and prognosis also differ according to glioma subtype (Citation3). At present, the primary treatment for glioma is surgical resection, usually combined with radiotherapy and chemotherapy (Citation4,Citation5). In recent years, there has been remarkable progress in glioma diagnosis, imaging, radiotherapy, chemotherapy, and neurosurgery; however, because of high rates of tumor recurrence, the poor prognosis of patients with malignant glioma has not been substantially improved (Citation6,Citation7). The median survival time of patients with glioblastoma is less than 14 months, and the one-year cumulative survival rate is less than 30% (Citation8,Citation9).
At present, the etiology of glioma is unclear, with potential risk factors including genetic factors, or specific gene polymorphisms, ionizing radiation, carcinogens of the nervous system, and virus infection, among others (Citation3). Previous studies demonstrated that, compared with patients with an ins/ins genotype at GAS5, those with ins/del or del/del genotypes have significantly increased glioma risk (Citation10). Further, EGF (Citation11), CCND1 c.870G>A (Citation12), pre-mir-146a (Citation13), microRNA-196a (Citation14), IL-12 (Citation3), and ERCC1 (Citation15) have clear effects on glioma risk. Zhang et al. identified a significant association between the XRCC1 Arg399Gln polymorphism and glioma risk specific to the Asian population (Citation16). Nevertheless, current knowledge cannot fully explain the causes of glioma, and further study of the effects of other gene polymorphisms on the risk of glioma is required (Citation17).
The CMYC protooncogene encodes an important transcription factor, which contributes to regulation of approximately 15% of human genes; participates in fundamental processes, including cell proliferation, metabolism, and differentiation (Citation18,Citation19); and is significantly up-regulated in numerous types of tumor (Citation20–22). CMYC is up-regulated in patients with cholangiocarcinoma (CCA), and this can increase the conversion of glucose to serine and reduce levels of mitochondrial oxidative phosphorylation, resulting in poor patient survival and prognosis (Citation20). In patients with pancreatic cancer, PRMT5 promotes the proliferation and aerobic glycolysis of pancreatic cancer cells by silencing the expression of the tumor suppressor, FBW7, thereby increasing levels of CMYC (Citation23). Further, CMYC expression is crucial to the metabolism and functional responses of natural killer cells (Citation24). In addition, the CMYC/EMT axis can inhibit nasopharyngeal carcinoma metastasis, under the regulation of hsa-miR-24, thus improving the survival rate and prognosis of patients with this type of cancer (Citation25). CMYC is also related to thyroid cancer (Citation26); however, the relationship between CMYC gene polymorphism and glioma risk has not been studied.
At present, the etiology of glioma is unclear, and the functional roles of single nucleotide polymorphisms (SNPs) in glioma pathogenesis have not been studied. Therefore, the molecular mechanisms underlying glioma require further study and more sensitive and specific early biomarkers are needed to improve the quality of life of patients with glioma. Considering the important role of CMYC in the carcinogenesis of many types of tumor, we explored the relationship between CMYC gene polymorphisms and susceptibility to glioma in Chinese children.
Materials and methods
Study subjects
A total of 190 patients with glioma and 248 healthy controls were included in this study. All subjects were treated at Guangzhou Women and Children’s Medical Center and The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University. The inclusion and exclusion criteria for subjects were described in detail in our previous reports (Citation27,Citation28). The case and control groups were matched for age and sex, and subjects were not close relatives. Before collection of blood samples, we discussed the study with parents or guardians of all participants and obtained their written informed consent. This study was approved by the Institutional Review Board of the centers in Wenzhou and Guangzhou.
Polymorphism selection and genotyping
Two potential functional polymorphisms of CMYC (rs4645943 C>T and rs2070583 A>G) were selected from the NCBI dbSNP database (http://www.ncbi.nlm.nih.gov/projects/SNP) and SNPinfo (http://snpinfo.niehs.nih.gov/snpfunc.htm) (Citation29–31). rs4645943 may be linked to multiple SNPs. rs2070583 may act as a miRNA binding site. Genomic DNA samples were extracted from venous blood, and a TaqMan SNP genotyping assay conducted on an Applied Biosystems 7900 real-time PCR system. More detailed SNP selection and genotyping criteria were described in our previous reports (Citation32–34).
Statistical analysis
We used the χ2 test to analyze differences in the distribution of characteristics between the case and control groups. A goodness-of-fit chi-squared test was used to evaluate Hardy-Weinberg equilibrium (HWE) in the control group. Crude and adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were obtained by logistic regression analysis to detect any association with glioma risk. In addition, we also conducted stratified analysis to evaluate correlations between age, sex, and clinical stage with the risk of glioma. P < 0.05 was considered to indicate a significant result. All analyses in this study were conducted using SAS 9.1 (SAS Institute, Cary, NC, USA).
Results
Characteristics of the study population
In this study, we genotyped 190 cases and 248 controls from Wenzhou and Guangzhou. Specific demographic characteristics are presented in detail in Table S1.
Association between CMYC gene polymorphisms and glioma susceptibility
and summarizes the genotype and allele frequencies of the selected polymorphisms in this case-control study and their relationship with the risk of glioma. In the control group, the genotype frequencies of the selected polymorphisms were all in accordance with Hardy–Weinberg equilibrium (HWE = 0.528 and 0.678 for rs4645943 C>T and rs2070583 A>G, respectively). In single locus analysis, we did not observe any statistically significant association between the frequencies of the two SNPs and risk of glioma. For rs4645943, compared with CC genotype carriers, CT genotype carriers (OR = 0.92, 95% CI = 0.62–1.37 p = 0.688) or TT genotype carriers (OR = 0.76, 95% CI = 0.35–1.64, p = 0.481) had no significant correlation with glioma risk. In addition, there was no significant association between rs4645943 and glioma risk in additive (OR = 0.90, 95% CI = 0.66–1.22 p = 0.486), dominant (OR = 0.91, 95% CI = 0.62–1.33 p = 0.608), or recessive models (OR = 0.80, 95% CI = 0.38–1.69 p = 0.555). For rs2070583, compared with AA genotype carriers, AG genotype carriers (OR = 0.98, 95% CI = 0.63–1.52 p = 0.922) or GG genotype carriers (OR = 0.62, 95% CI = 0.16–2.46, p = 0.495) had no significant correlation with glioma risk. In addition, there was no significant association between rs4645943 and glioma risk in additive (OR = 0.91, 95% CI = 0.62–1.33 p = 0.631), dominant (OR = 0.94, 95% CI = 0.62–1.44 p = 0.783), or recessive models (OR = 0.61, 95% CI = 0.15–2.40 p = 0.477).
Table 1. Genotype frequencies of CMYC polymorphisms in cases and controls.
Table 2. Association between CMYC gene polymorphisms and glioma susceptibility in Chinese children.
Stratified analysis
We also conducted stratified analyses according to age, sex, glioma subtype, and clinical stage (); however, no significant associations were detected.
Table 3. Stratification analysis of risk genotypes with glioma susceptibility.
Haplotype analysis
As shown in the , we can see that none of the results are statistically significant.
Table 4. The frequency of inferred haplotypes of CMYC gene based on observed genotypes and their association with the risk of Glioma.
Discussion
In this case-control study, we analyzed the association between two potential functional polymorphisms of CMYC and glioma risk in children. No association of either selected SNP (rs4645943 C>T and rs2070583 A>G) and glioma risk was detected in either whole cohort or stratified analyses.
The CMYC oncogene encodes a helix-loop-helix leucine zipper transcription factor, which is dysregulated in numerous cancers, and has differing effects on the growth, proliferation, metastasis, and angiogenesis of cancer cells (Citation35,Citation36). CMYC protein levels are strictly regulated by amino acids, and the transcription factor is very important for the metabolic and functional responses induced by IL-2/IL-12 [24]. Liu et al. found that CMYC and TIP110 can regulate the expression of one another, thereby playing an important role in hematopoiesis regulation (Citation37). Chen et al. reported that CMYC can regulate miR-200c, to inhibit the tumor suppressor gene, PTEN, leading to cancer occurrence and progression (Citation35). Further, Zhong et al. determined that CMYC can interact with SREBP2 and synergistically activate the expression of HMGCR, thus promoting the growth and migration of esophageal squamous cell carcinoma cells (Citation38), while Luo et al. found that CMYC protein levels in human CCA cells were significantly higher than those in normal bile duct epithelial cells, and that expression of CMYC in human CCA cells could eliminate contact inhibition and promote their invasion and migration (Citation39). Moreover, Zaharieva et al. discovered that changes in CMYC are related to low survival rate in patients with bladder cancer, with increased expression and amplification of CMYC closely related to development of late stage bladder cancer (Citation40).
In our previous research, we determined that, compared with patients carrying the CT/TT genotypes of rs4645943 in CMYC, those with the CC genotype had a significantly reduced risk of neuroblastoma (Citation30); however, no significant association was found between CMYC gene polymorphism and Wilms tumor risk (Citation29).
Berberich et al. reported that CMYC can increase the resistance of glioblastoma cells to alectinib (Citation41), while Odia et al. found that the CMYC protein has an adverse prognostic role in malignant glioma expressing mutant IDH1 (Citation42); however, there are no reported investigations of the relationship between CMYC gene polymorphisms and glioma risk. As the CMYC gene has important roles in various cancers, we collected blood samples from glioma cases and healthy controls to study the association between CMYC gene polymorphisms and glioma risk. Our results did not detect any significant association between CMYC polymorphisms and glioma risk, although this may be attributable to the low sample size included in this study.
Although we are the first group to study the association between CMYC gene polymorphisms and glioma risk, there remains scope for more comprehensive investigations. Further, while this was a multi-center study, due to the low incidence of glioma and the small number of cooperating hospitals, the sample size we collected was far from sufficient, potentially leading to some anomalies in our results. In addition, all subjects were from Wenzhou and Guangzhou; hence it may be inappropriate to apply our conclusions to patients from other regions and ethnic groups. Moreover, only two SNPs were analyzed in our research, and some important potentially functional SNPs may have been omitted. Finally, we did not investigate the relationships of gene-gene or gene-environment interactions with glioma risk. Nevertheless, our data indicate that CMYC may not be an independent factor influencing the risk of glioma.
Conclusions
In summary, our study did not find a significant correlation between CMYC gene polymorphisms and glioma risk. In the future, we will conduct further research based on a larger sample collection.
Supplemental Material
Download PDF (96.5 KB)Acknowledgments
This study was supported by grants from the National Natural Science Foundation of China [No: 81802346].
Disclosure statement
The authors declare no conflicts of interest.
Data availability statement
All the data used to support the findings of this study are available from the corresponding author upon request.
Supplementary materials
Supplemental Table 1: frequency distribution of selected variables for Glioma cases and cancer-free controls. (Supplementary Materials)
References
- Wang H, Wu J, Guo W. SP1-Mediated Upregulation of lncRNA LINC01614 Functions a ceRNA for miR-383 to Facilitate Glioma Progression Through Regulation of ADAM12. Onco Targets Ther. 2020;13:4305–18. doi:https://doi.org/10.2147/OTT.S242854.
- Li X-Y, Huang G-H, Liu Q-K, Yang X-T, Wang K, Luo W-Z, et al. Porf-2 Inhibits Tumor Cell Migration Through the MMP-2/9 Signaling Pathway in Neuroblastoma and Glioma. Front Oncol. 2020;10:975. doi:https://doi.org/10.3389/fonc.2020.00975.
- Zhao B, Meng L-Q, Huang H-N, Pan Y, Xu Q-Q. A novel functional polymorphism, 16974 A/C, in the interleukin-12-3' untranslated region is associated with risk of glioma. DNA Cell Biol. 2009;28(7):335–41. doi:https://doi.org/10.1089/dna.2008.0845.
- Chang C-Y, Pan P-H, Li J-R, Ou Y-C, Wang J-D, Liao S-L, et al. Aspirin induced glioma apoptosis through Noxa Upregulation. Int J Mol Sci. 2020;21(12):4219. doi:https://doi.org/10.3390/ijms21124219.
- Zhang Q, Zhong H, Fan Y, Liu Q, Song J, Yao S, et al. Immune and Clinical Features of CD96 Expression in Glioma by in silico Analysis. Front Bioeng Biotechnol. 2020;8:592. doi:https://doi.org/10.3389/fbioe.2020.00592.
- Wang F, Zhou Y, Cheng S, Lou J, Zhang X, He Q, et al. Gint4.T-Modified DNA Tetrahedrons Loaded with Doxorubicin Inhibits Glioma Cell Proliferation by Targeting PDGFRβ. Nanoscale Res Lett. 2020;15(1):150. doi:https://doi.org/10.1186/s11671-020-03377-y.
- Xiao L, Li X, Mu Z, Zhou J, Zhou P, Xie C, et al. FTO inhibition enhances the Antitumor Effect of Temozolomide by Targeting MYC-miR-155/23a Cluster-MXI1 Feedback Circuit in Glioma . Cancer Res. 2020;80(18):3945–58. doi:https://doi.org/10.1158/0008-5472.
- Cai H, Liu W, Liu X, Li Z, Feng T, Xue Y, et al. Advances and prospects of vasculogenic mimicry in glioma: a potential new therapeutic target? Onco Targets Ther. 2020;13:4473–83. doi:https://doi.org/10.2147/OTT.S247855.
- Li Y, et al. Downregulation of LUZP2 is correlated with poor prognosis of low-grade glioma. Biomed Res Int. 2020;2020:9716720.
- Yuan J, Zhang N, Zheng Y, Chen Y-D, Liu J, Yang M, et al. LncRNA GAS5 indel genetic polymorphism contributes to glioma risk through interfering binding of transcriptional factor TFAP2A. DNA Cell Biol. 2018;37(9):750–7. doi:https://doi.org/10.1089/dna.2018.4215.
- da Silveira F. d C A, Lopes B. d A, da Fonseca CO, Quirico-Santos T, de Palmer Paixão ICN, de Amorim L. M d F, et al. Analysis of EGF + 61A>G polymorphism and EGF serum levels in Brazilian glioma patients treated with perillyl alcohol-based therapy. J Cancer Res Clin Oncol. 2012;138(8):1347–54. doi:https://doi.org/10.1007/s00432-012-1203-5.
- Chen X, Zhao T, Li L, Xu C, Zhang X, Tse V, et al. CCND1 G870A polymorphism with altered cyclin D1 transcripts expression is associated with the risk of glioma in a Chinese population. DNA Cell Biol. 2012;31(6):1107–13. doi:https://doi.org/10.1089/dna.2011.1521.
- Permuth-Wey J, Thompson RC, Burton Nabors L, Olson JJ, Browning JE, Madden MH, et al. A functional polymorphism in the pre-miR-146a gene is associated with risk and prognosis in adult glioma. J Neurooncol. 2011;105(3):639–46. doi:https://doi.org/10.1007/s11060-011-0634-1.
- Dou T, Wu Q, Chen X, Ribas J, Ni X, Tang C, et al. A polymorphism of microRNA196a genome region was associated with decreased risk of glioma in Chinese population. J Cancer Res Clin Oncol. 2010;136(12):1853–9. doi:https://doi.org/10.1007/s00432-010-0844-5.
- Chen P, Wiencke J, Aldape K, Kesler-Diaz A, Miike R, Kelsey K, et al. Association of an ERCC1 polymorphism with adult-onset glioma. Cancer Epidemiol Biomarkers Prev. 2000;9(8):843–7.
- Zhang L, Qiu Z, Luo J, Shu W. X-ray repair cross-complementing gene 1 Arg399Gln polymorphism and glioma risk among Asians: A meta-analysis based on 2 326 cases and 3 610 controls. Neural Regen Res. 2012;7(29):2313–9. doi:https://doi.org/10.3969/j.issn.1673-5374.2012.29.011.
- Xu H, Chai S, Wang Y, Wang J, Xiao D, Li J, et al. Molecular and clinical characterization of PARP9 in gliomas: A potential immunotherapeutic target. CNS Neurosci Ther. 2020;26(8):804–14. doi:https://doi.org/10.1111/cns.13380.
- Zhang J, Zhou L, Nan Z, Yuan Q, Wen J, Xu M, et al. Knockdown of c‐Myc activates Fas-mediated apoptosis and sensitizes A549 cells to radiation . Oncol Rep. 2017;38(4):2471–9. doi:https://doi.org/10.3892/or.2017.5897.
- Dong H, Adams NM, Xu Y, Cao J, Allan DSJ, Carlyle JR, et al. The IRE1 endoplasmic reticulum stress sensor activates natural killer cell immunity in part by regulating c-Myc. Nat Immunol. 2019;20(7):865–78. doi:https://doi.org/10.1038/s41590-019-0388-z.
- Xu L, Wang L, Zhou L, Dorfman RG, Pan Y, Tang D, et al. The SIRT2/cMYC Pathway Inhibits Peroxidation-Related Apoptosis In Cholangiocarcinoma Through Metabolic Reprogramming. Neoplasia. 2019;21(5):429–41. doi:https://doi.org/10.1016/j.neo.2019.03.002.
- Basit F, Andersson M, Hultquist A. The Myc/Max/Mxd Network Is a Target of Mutated Flt3 Signaling in Hematopoietic Stem Cells in Flt3-ITD-Induced Myeloproliferative Disease. Stem Cells Int. 2018;2018:3286949. doi:https://doi.org/10.1155/2018/3286949.
- Lee Y-S, Heo W, Son C-H, Kang C-D, Park Y-S, Bae J, et al. Upregulation of Myc promotes the evasion of NK cell‐mediated immunity through suppression of NKG2D ligands in K562 cells. Mol Med Rep. 2019;20(4):3301–7. doi:https://doi.org/10.3892/mmr.2019.10583.
- Qin Y, Hu Q, Xu J, Ji S, Dai W, Liu W, et al. PRMT5 enhances tumorigenicity and glycolysis in pancreatic cancer via the FBW7/cMyc axis. Cell Commun Signal. 2019;17(1):30. doi:https://doi.org/10.1186/s12964-019-0344-4.
- Loftus RM, Assmann N, Kedia-Mehta N, O'Brien KL, Garcia A, Gillespie C, et al. Amino acid-dependent cMyc expression is essential for NK cell metabolic and functional responses in mice. Nat Commun. 2018;9(1):2341. doi:https://doi.org/10.1038/s41467-018-04719-2.
- Su B, Xu T, Bruce JP, Yip KW, Zhang N, Huang Z, et al. hsa‐miR‐24 suppresses metastasis in nasopharyngeal carcinoma by regulating the c‐Myc/epithelial‐mesenchymal transition axis. Oncol Rep. 2018;40(5):2536–46. doi:https://doi.org/10.3892/or.2018.6690.
- Sakr HI, Chute DJ, Nasr C, Sturgis CD. cMYC expression in thyroid follicular cell-derived carcinomas: a role in thyroid tumorigenesis. Diagn Pathol. 2017;12(1):71. doi:https://doi.org/10.1186/s13000-017-0661-0.
- Huang X, Zhao J, Fu W, Zhu J, Lou S, Tian X, et al. The association of RAN and RANBP2 gene polymerphisms with Wilms tumor risk in Chinese children. J Cancer. 2020;11(4):804–9. doi:https://doi.org/10.7150/jca.36651.
- Cheng J, Zhuo Z, Xin Y, Zhao P, Yang W, Zhou H, et al. Relevance of XPD polymorphisms to neuroblastoma risk in Chinese children: a four-center case-control study. Aging. 2018;10(8):1989–2000. doi:https://doi.org/10.18632/aging.101522.
- Liu J, Hua R-X, Fu W, Zhu J, Jia W, Zhang J, et al. MYC gene associated polymorphisms and Wilms tumor risk in Chinese children: a four-center case-control study. Ann Transl Med. 2019;7(18):475. doi:https://doi.org/10.21037/atm.2019.08.31.
- Pan J, Zhu J, Wang M, Yang T, Hu C, Yang J, et al. Association of MYC gene polymorphisms with neuroblastoma risk in Chinese children: A four-center case-control study. J Gene Med. 2020;22(8):e3190. doi:https://doi.org/10.1002/jgm.3190.
- Yang T, Wen Y, Li J, Tan T, Yang J, Pan J, et al. Association of CMYC polymorphisms with hepatoblastoma risk. Transl Cancer Res Tcr. 2020;9(2):849–55. doi:https://doi.org/10.21037/tcr.2019.12.19.
- Wang J, Zhuo Z, Chen M, Zhu J, Zhao J, Zhang J, et al. RAN/RANBP2 polymorphisms and neuroblastoma risk in Chinese children: a three-center case-control study. Aging. 2018;10(4):808–18. doi:https://doi.org/10.18632/aging.101429.
- Hua R-X, Zhuo Z, Ge L, Zhu J, Yuan L, Chen C, et al. LIN28A gene polymorphisms modify neuroblastoma susceptibility: A four-centre case-control study. J Cell Mol Med. 2020;24(1):1059–66. doi:https://doi.org/10.1111/jcmm.14827.
- Zhang Z, Yin J, Xu Q, Shi J. Association between the XPG gene rs2094258 polymorphism and risk of gastric cancer. J Clin Lab Anal. 2018;32(8):e22564. doi:https://doi.org/10.1002/jcla.22564.
- Chen P, Guo X, Zhang L, Zhang W, Zhou Q, Tian Z, et al. MiR-200c is a cMyc-activated miRNA that promotes nasopharyngeal carcinoma by downregulating PTEN. Oncotarget. 2017;8(3):5206–18. doi:https://doi.org/10.18632/oncotarget.14123.
- Day AH, Übler MH, Best HL, Lloyd-Evans E, Mart RJ, Fallis IA, et al. Targeted cell imaging properties of a deep red luminescent iridium(iii) complex conjugated with a c-Myc signal peptide. Chem Sci. 2020;11(6):1599–606. doi:https://doi.org/10.1039/c9sc05568a.
- Liu Y, Timani K, Mantel C, Fan Y, Hangoc G, Cooper S, et al. TIP110/p110nrb/SART3/p110 regulation of hematopoiesis through CMYC. Blood. 2011;117(21):5643–51. doi:https://doi.org/10.1182/blood-2010-12-325332.
- Zhong C, Fan L, Li Z, Yao F, Zhao H. SREBP2 is upregulated in esophageal squamous cell carcinoma and co‐operates with c‐Myc to regulate HMGCR expression. Mol Med Rep. 2019;20(4):3003–10. doi:https://doi.org/10.3892/mmr.2019.10577.
- Luo G, Li B, Duan C, Cheng Y, Xiao B, Yao F, et al. c‐Myc promotes cholangiocarcinoma cells to overcome contact inhibition via the mTOR pathway. Oncol Rep. 2017;38(4):2498–506. doi:https://doi.org/10.3892/or.2017.5913.
- Zaharieva B, Simon R, Ruiz C, Oeggerli M, Mihatsch MJ, Gasser T, et al. High-throughput tissue microarray analysis of CMYC amplificationin urinary bladder cancer. Int J Cancer. 2005;117(6):952–6. doi:https://doi.org/10.1002/ijc.21253.
- Berberich A, Schmitt L-M, Pusch S, Hielscher T, Rübmann P, Hucke N, et al. cMyc and ERK activity are associated with resistance to ALK inhibitory treatment in glioblastoma. J Neurooncol. 2020;146(1):9–23. doi:https://doi.org/10.1007/s11060-019-03348-z.
- Odia Y, Orr BA, Bell WR, Eberhart CG, Rodriguez FJ. cMYC expression in infiltrating gliomas: associations with IDH1 mutations, clinicopathologic features and outcome. J Neurooncol. 2013;115(2):249–59. doi:https://doi.org/10.1007/s11060-013-1221-4.