Abstract
Background
Increasing evidence recognizes that DNA methylation abnormalities play critical roles in cancer development. Our previous genome-wide methylation profile showed that tumor necrosis factor-alpha-induced protein 8 like 3 (TIPE3) was hypermethylated in nasopharyngeal carcinoma (NPC). However, the relationship between TIPE3 methylation and its mRNA expression, as well as its biological roles in NPC are unknown.
Methods
Bisulfite pyrosequencing and quantitative RT-PCR were performed to quantify the TIPE3 methylation and expression levels. Kaplan-Meier curves and Cox regression analysis were used to estimate the correlation between TIPE3 methylation levels and survival in two patient cohorts collected from two hospitals (n = 441). The MTT, colony formation, Transwell migration and invasion assays, and xenograft tumor growth and lung metastatic colonization models were used to identify the functions of TIPE3 on NPC cells.
Results
We found that TIPE3 CpG island (CGI) was hypermethylated and its mRNA levels were downregulated in many cancers, including NPC. TIPE3 downregulation was associated with its CGI hypermethylation. Furthermore, NPC patients with high TIPE3 CGI methylation levels had poorer clinical outcomes than those with low methylation levels. The TIPE3 CGI methylation level was an independent prognostic factor. Moreover, restoring TIPE3 expression significantly inhibited NPC cell proliferation, migration and invasion in vitro, and suppressed tumor growth and lung metastatic colonization in vivo, while silencing TIPE3 acted in an opposite way.
Conclusions
TIPE3 downregulation correlates with its CGI hypermethylation in several solid cancers. TIPE3 acts as a tumor suppressor in NPC, providing a further insight into NPC progression and representing a potential prognostic biomarker for NPC.
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Background
Nasopharyngeal carcinoma (NPC) is a common head and neck cancer arising from the nasopharynx epithelium, with the highest prevalence in southern China [1, 14, 24]. Recently, the TIPE family members have been recognized as inflammation, immunity, and cancer regulators [25, 26]. TIPE, the first identified member of this family, can regulate apoptosis and promote tumor metastasis and proliferation [27, 28]. TIPE1 was reported to be essential for TNF-α-induced cell death [29]. TIPE2 can maintain immune hemostasis and function as a tumor suppressor [30, 31]. TIPE3 was found to be upregulated in lung cancer, esophageal cancer, cervical cancer, and colon adenocarcinoma. The unique NT region of TIPE3, which is not seen in other members of the TIPE family, is believed to be responsible for its unique ability to promote cell growth and survival. Furthermore, TIPE3 lacking the NT region appeared to exert a tumor suppression effect [14]. In the present study, we found that ectopic expression of TIPE3 significantly suppressed NPC cell proliferation and invasion in vitro and in vivo, indicating that TIPE3 might act as a tumor suppressor in NPC and play a dual role in cancer progression. In fact, many genes are reported to act as either tumor suppressors or oncogenes in different cancer types. Nevertheless, the underlying mechanisms for these contradictory roles of TIPE3 in different cancers remain to be determined.
At present, the clinical decision making for NPC patients mainly relies on the TNM staging system [3], even though it cannot accurately select those at high risk of treatment failure. Over the past decades, numerous studies have focused on develo** efficient prognostic molecular biomarkers, such as EBV-DNA, miRNAs, and gene expression [32,33,34]. However, there is still no effective predictive model for NPC patients. Recently, increasing evidences has demonstrated that both the single-gene loci and the genome-wide profiling indicated a strong potential to predict outcomes in malignant tumors. For single-gene loci, hypermethylation of CDKN2A in colorectal cancer, MGMT in glioblastoma, BRCA1 in breast cancer were reported to be associated with poor clinical outcomes [35,36,37]. For genome-wide profiling, the methylation gene panel as a prognostic biomarker of prostate, lung, and other cancers have also been identified [38, 39]. We previously constructed a six-hypermethylated gene panel to predict NPC patients’ survival [12]. However, the clinical applications of several aberrantly methylated genes in NPC remain unknown. In this study, our findings demonstrated that NPC patients with high TIPE3 CGI methylation level exhibited a significantly shorter OS, DFS, and DMFS compared with patients with low methylation level. These results implied that the TIPE3 CGI methylation level could help to identify a subgroup of patients with high risk of treatment failure and guide more individualized therapy.
Conclusions
This study demonstrated that TIPE3 mRNA downregulation was correlated with its CGI hypermethylation in human solid cancers. NPC patients with low TIPE3 CGI methylation levels were at low risk of treatment failure, which might be caused by the tumor suppression effects of TIPE3. Therefore, TIPE3 is a potential novel prognostic biomarker and therapeutic target for NPC patients.
Abbreviations
- AJCC:
-
American joint committee on cancer
- CGI:
-
CpG island
- CLL:
-
Chronic lymphocytic leukemia.
- DFS:
-
Disease-free survival
- DMFS:
-
Distant metastasis-free survival
- EA-IgA:
-
Early antigen immunoglobulin A
- EBV:
-
Epstein-Barr virus
- GAPDH:
-
Glyceraldehyde-3-phosphate dehydrogenase
- LDH:
-
Lactate dehydrogenase
- NCCN:
-
National comprehensive cancer network
- NPC:
-
Nasopharyngeal carcinoma
- NPEC:
-
Normal nasopharyngeal epithelial cell
- NT:
-
N-terminal
- OS:
-
Overall survival
- ROC:
-
Receiver operating characteristic
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
- TCGA:
-
The Cancer Genome Atlas
- TIPE3:
-
Tumor necrosis factor-alpha-induced protein 8 like 3
- TNM:
-
Tumor-node-metastasis
- VCA-IgA:
-
Viral capsid antigen immunoglobulin A
- WHO:
-
World health organization
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Funding
This work was supported by grants from the Natural Science Foundation of Guangdong Province (2018B030306045); the Young Teachers Cultivation Project of Sun Yat-sen University (16ykpy21); the National Natural Science Foundation of China (81572658, 81702700); the Young Elite Scientist Sponsorship Program by CAST (2017QNRC001); and the Open Project of State Key Laboratory of Oncology in South China (HN2017–03). The funders had no role in the study design, data collection, analysis, decision to publish or the preparation of the manuscript.
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NL and JM designed the research. XYR, XW, YQL, JZ, QMH, XJY, XRT, YQW, and PPZ conducted the experiments and analyzed the data. JM, NL, XZC, and BC provided the reagents. JM, NL, XYR, XW and YQL wrote the manuscript. All authors read and approved the final manuscript.
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This research was authorized by the Institutional Ethical Review Boards of both hospitals, and written informed consents were provided by all patients for using their biopsies. The animal experiments were approved by the Animal Care and Use Ethnic Committee of our Center.
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Additional files
Additional file 1:
Table S1. The methylation probes of TIPE3 in Infinium Human Methylation 450 K BeadChip (Illumina). (XLSX 16 kb)
Additional file 2:
Table S2. Correlations between TIPE3 methylation levels and clinical features in patients with nasopharyngeal carcinoma from the training and validation cohorts. (DOCX 24 kb)
Additional file 3:
Table S3. Univariate Cox regression analyses of the significant of different prognostic variables in nasopharyngeal carcinoma. (DOCX 19 kb)
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Ren, XY., Wen, X., Li, YQ. et al. TIPE3 hypermethylation correlates with worse prognosis and promotes tumor progression in nasopharyngeal carcinoma. J Exp Clin Cancer Res 37, 227 (2018). https://doi.org/10.1186/s13046-018-0881-5
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DOI: https://doi.org/10.1186/s13046-018-0881-5