IRAK2, an immune and radiation-response gene, correlates with advanced disease features but predicts high post-irradiation local control in resected oral cancer patients

doi:10.21203/rs.3.rs-2434157/v1

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Research Article

IRAK2, an immune and radiation-response gene, correlates with advanced disease features but predicts high post-irradiation local control in resected oral cancer patients

https://doi.org/10.21203/rs.3.rs-2434157/v1

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Radiotherapy (RT) is an effective treatment modality for managing resected oral squamous cell carcinoma (OSCC) patients. The present study conducted gene ontology to explore the functional biological analysis of IRAK2 and performed case analysis to define its clinical role in disease progression and mediating tumor response to RT. Transcriptome sequencing data from the OML1 oral cancer cell line was analyzed to explore IR-responding genes. For identified significant IR-responsive genes, gene ontology (GO) analysis was conducted for exploring their involved bio-logical processes. From Jan. 2007 to Dec. 2014, 172 OSCC patients who received radical surgery with (n = 93) and without (n = 79) postoperative RT were identified for clinical validation. Their OSCC formalin-fixed paraffin-embedded histological samples were examined retrospectively to define IRAK2 expression. After irradiation treatments, IRAK2 upregulation was identified obviously suggesting its role as a potential radiation biomarker. Moreover, GO enrichment analysis showed IRAK2 involved ten of 14 top enriched post-irradiation biological processes, focusing on stress response and immune modulation in vitro. Clinically, high IRAK2 expression was correlated with several adverse disease features, including pT3-4 status (P = 0.01), advanced overall stage (P = 0.02), and positive bone invasion (P = 0.01). In patients who underwent RT, the IRAK2-high group was associated with fewer post-irradiation local recurrence (multivariate HR, 0.243; 95% CI, 0.071–0.838; P = 0.025) than the IRAK2-low group. However, the local control benefit did not translate into patient survival. On the other hand, for patients with no RT, IRAK2 did not predict local control. IRAK2 plays a crucial role in the radiation-induced response. In summary, patients with high IRAK2 expression demonstrated more advanced disease features but predicted higher post-irradiation local control than those with low expression. These findings support IRAK2 as a potential predictive biomarker for RT response in OSCC patients. Further prospective clinical studies are encouraged.

Oral squamous cell carcinoma (OSCC) ranks sixth in cancer incidence in Taiwan, and radiotherapy (RT) is one of the essential treatment modalities for managing OSCC patients[1]. RT is mainly applied in conjunction with surgery in advanced-stage patients[1, 2]. However, local recurrence and distant metastasis after RT are still major clinical problems that lead to poor overall survival (OS) in oral cancer patients[2, 3]. Although some clinical and pathologic factors help identify high-risk patients in developing cancer recurrence, there is a need to identify reliable molecular biomarkers to predict tumor response and clinical outcomes in irradiated OSCC patients.

Interleukin-1 receptor-associated kinase 2 (IRAK2) is a critical regulator of interleukin-1 receptor (IL-1R)/toll-like receptor (TLR) -mediated inflammation, involved in the NF-kB and MAPK signaling pathway[4]. IRAK2 is also recognized to participate in the apoptotic response through multiple mechanisms[5–7]. Our previous study found that IRAK2 expresses low levels in radioresistant OSCC cell lines, whereas overexpression of IRAK2 restored the radiosensitivity via modulating caspase 8/3-mediated apoptosis, suggesting that IRAK2 is a radiation-responding gene that is potentially involved in tumor response to ionizing radiation IR[7]. Furthermore, we observed that early-stage OSCC patients with higher IRAK2 expression demonstrate better post-radiation local control than patients with lower expression[7]. However, the clinical significance of IRAK2 associated with RT is still not fully understood for the whole group of oral cancer patients.

As a result, the present study expanded prior RNA sequencing-based data to functional biological analysis for further verifying the molecular functions of IRAK2 that may exhibit changes correlating with radiosensitivity. Moreover, we sought to analyze associations between IRAK2 expression and post-RT outcomes.

Clinical specimens

As mentioned previously[8], postoperative RT with or without concurrent Cisplatin-based chemotherapy was prescribed for resected oral cancer patients with indicated adverse features. RT indications were pT3-4, pN+, and positive or close surgical margins (i.e., ≤ 1 mm). Briefly, we used Volumetric-modulated Arc Therapy (VMAT) to prescribe irradiation with doses as follows: 60–72 Gy to the primary surgical bed, 60–66 Gy to the high-risk nodal basins, and 50–60 Gy to the low-risk nodal basins. Electronic portal imaging and cone-beam tomography were conducted weekly during the irradiation course. The conventionally fractionated dose was delivered, i.e., 1.8-2 Gy per day, five days per week, with 6-MV photons.

Cisplatin-based chemotherapy was concurrently given in patients with additional high-risk adverse features, such as positive/close surgical margins and extracapsular nodal extension, playing the role of radiosensitizer. The prescribed dose was 80–100 mg/m2 administered every three weeks, two to three cycles during RT, depending on the patient's condition. All OSCC formalin-fixed paraffin-embedded histological samples were examined retrospectively.

Cell culture and radiation treatment

OML1 cells were cultured as described previously[7, 9, 10] and maintained in RPMI1640 containing 10% FBS, 1% penicillin-streptomycin, and 2 mM L-glutamine. The cells were irradiated 4 Gy using a 6-MV medical linear accelerator (LINAC; Elekta, Crawley, Surrey, UK), as described previously[7].

Differential gene expression profile in response to radiation

We used RNA-seq data to study gene expression profiles of the radiation-induced transcriptional responses, as described previously[7]. These data had been deposited in the Gene Expression Omnibus (GEO) database (GSE165585).

The RNA-seq data from OML1 cells was conducted after 4-Gy radiation exposure or none. RPKM (Reads Per Kilobase of transcript per Million reads mapped) measured gene or transcript expression levels. Overall, 57,773 genes were obtained. Transcripts with zeroed RPKM values were excluded, and the filtered set of 29208 genes was used for the analyses outlined below. The genes that showed an absolute value of the log2 ratio of > 1.4 and an adjusted p-value of ≤ 0.01 were considered potential candidates. A volcano plot was used to depict the distribution of differentially expressed genes between irradiated and non-irradiated cells.

Functional analysis of radiation-responsive gene

Gene Ontology (GO) enrichment analysis was performed to classify the functions of significantly expressed genes by using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) (https://david.ncifcrf.gov/). Genes of enriched biological processes and gene ontology (GO) terms using the functional annotation tool were identified by following instructions of the DAVID manual.

Western Blotting

OML1 cells were seeded in 10 cm plates and treated with or without of radiation (2 or 4Gy) for 24 h. The cells were harvested and lysed, and protein concentrations were measured by using the Bio-Rad protein assay kit (Bio-Rad, Richmond, CA, USA). Equal amounts of protein (50 µg) were loaded onto 12.5% SDS-polyamide gel electrophoresis and then transferred to PVDF membranes. After transfer, the membranes were blocked with 5% non-fat milk in PBS containing 0.1% Tween 20. The membrane was blotted with IRAK2 antibody (Cell Signaling Technology, Beverly, MA) and incubated with a horseradish peroxidase-labeled secondary antibody. The protein band was visualized using a chemiluminescence (ECL) detection kit (Millipore).

Immunohistochemistry

The IRAK2 monoclonal antibody (dilution 1:50) was purchased from Abnova (Taipei, Taiwan). We examined all OSCC formalin-fixed paraffin-embedded histological samples retrospectively. Antigen retrieval was conducted by using 4-µm–thick Paraffin-embedded oral tumor sections. We used the Super Sensitive™ Polymer-HRP IHC detection system (Biogenex, San Ramon, CA) to measure the IHC stained levels. Oncological pathologists defined protein expression scores by using the multiplied values of stained intensity (0–3) and percentages (0-100%), rating 0-300. Due to different clinical and biological features, we used cutoff points of 110 in early and 75 in advanced OSCC patient samples to classify levels of IRAK2 protein expression.

Research Database of Clinical Outcomes

Data were collected through the Prospective-Coding Cancer Registry Database in Dalin, Taiwan, a regular national-audit cancer database for oncological statistics and research. Patient features were analyzed according to relevant demographic and clinical factors, such as age, clinical and pathologic stage, surgical margin, RT dose, and chemotherapy. Pearson correlation was applied to evaluate correlations between clinical-pathological parameters and IRAK2 expression.

Statistical Analysis

All statistical analyses were performed using the SigmaPlot software, version 10.0 (Systat Software Inc., San Jose, CA, USA) and SPSS (version 12.0; SPSS Inc., Chicago, IL, USA). Continuous data were presented as mean ± standard deviation, and their statistically significant levels were calculated using the Student’s t-test. Categorical data were analyzed using the Chi-square test. Time-to-event endpoints were estimated using the Kaplan-Meier plot, and the log-rank test was applied to assess curve differences between groups. Cox proportional hazards regression model was applied for univariate and multivariate analysis. All hazard ratios were provided with 95% confidence intervals to delineate effective size. P values of less than 0.05 were defined as statistical significance.

Investigating irradiation-response genes in irradiated OML1 oral cancer cells through transcriptome analysis

Results of comprehensive genomic profiling in our previous study showed that compared to parental OML1 and radioresistant OML1-R cells, several genes are strongly upregulated after radiation treatment[7], suggesting their potential roles in response to radiotherapy. The present study utilized these transcriptomic data and focused on the post-irradiation upregulated gene profiles in OML1 cells. Figure 1 shows the distribution of gene expression between untreated and IR-treated OML1 cells. After irradiation, 25 genes were significantly upregulated based on the selection criterion of log2 fold change (FC) of > 1.4 with a p-value of < 0.01 (Fig. 1).

Gene ontology analysis shows enriched biological processes of post-irradiation upregulated genes

In accordance with RNA-seq data, Fig. 2a depicts expression levels of the 25 post-IR upregulated genes, including PHLDA2, HBEGF, IRAK2, MCAM, CSF2, CA2, INHBA, LY6G5B, INHBB, HOXC13, AC007405.6, MEF2BNB-MEF2B, ACTR3C, HSD3B7, ECM1, RP11-81N13.1, AC074091.13, LYSMD1, FABP3, C15orf59, AC099342.1, CD74, C4B, FRMD3, and KIAA0408. Furthermore, we conducted western blot at 24 h after irradiation (0, 2 and 4 Gy) in OML1 cells. Compared with the control (0 Gy), IRAK2 protein expression increased with the increase of radiation dose (Fig. 2b). Gene Ontology (GO) analysis focused on these genes, for which the marked changes may predict biological processes. We observed the 25 genes involved in signaling various cell regulations, such as cell communication, migration, apoptosis, proliferation, and stress responses (e.g., chemicals, lipid, and oxygen). In addition, several genes were found to participate in the immune and inflammatory response, including cytokine-mediated signaling pathway and regulation of immune system processing (Fig. 3). Significant GO enrichment profiles in the biological process are summarized in Table 1. Note that IRAK2 plays multiple roles involving ten of the top 14 enriched GO biological processes, including immune regulation and stress response.

Table 1

The top 14 enriched gene-ontology (GO) terms of the 25 post-irradiation upregulated genes
GO ID and term	Count	P	Genes
0010647: positive regulation of cell communication	8	0.000190	CD74, ECM1, CSF2, IRAK2, CA2, INHBB, INHBA, HBEGF
0023056: positive regulation of signaling	8	0.000196	CD74, ECM1, CSF2, IRAK2, CA2, INHBB, INHBA, HBEGF
0042221: response to chemical	11	0.000696	C4B, CD74, HSD3B7, ECM1, FABP3, CSF2, IRAK2, CA2, INHBB, INHBA, HBEGF
0006950: response to stress	10	0.001559	C4B, CD74, ECM1, CSF2, IRAK2, CA2, MCAM, INHBB, INHBA, HBEGF
0045937: positive regulation of phosphate metabolic process	6	0.001669	CD74, FABP3, CSF2, IRAK2, INHBB, INHBA
0030334: regulation of cell migration	5	0.002614	CD74, ECM1, MCAM, PHLDA2, HBEGF
2001233: regulation of apoptotic signaling pathway	4	0.004272	CD74, CSF2, INHBB, INHBA
0002682: regulation of immune system process	6	0.005518	C4B, CD74, ECM1, IRAK2, CA2, INHBA
0048608: reproductive structure development	4	0.005576	CSF2, INHBB, INHBA, PHLDA2
0033993: response to lipid	5	0.005924	FABP3, CSF2, IRAK2, CA2, INHBA
0001959: regulation of cytokine-mediated signaling pathway	3	0.007031	CD74, ECM1, IRAK2
1901701: cellular response to oxygen-containing compound	5	0.007289	CSF2, IRAK2, CA2, INHBB, INHBA
0006952: defense response	6	0.008326	C4B, CD74, ECM1, IRAK2, INHBB, INHBA
0042127: regulation of cell proliferation	6	0.009138	CD74, ECM1, FABP3, CSF2, INHBA, HBEGF
Note that IRAK2 play multiple roles involving ten of 14 GO terms of biology processes. Abbreviation: GO, gene ontology

IRAK2 expression predicts local control for patients who received radiotherapy

Correlations between IRAK2 and the clinical features of OSCC patient samples were summarized in Table 2. The results indicated that higher expression of IRAK2 correlated statistically significantly with pT3-4 (P = 0.01), advanced pathology stage (P = 0.02), and positive bone invasion (P = 0.01). Among irradiated patients, the high IRAK2-expressed group showed better local control than those with low IRAK2 expression (Fig. 4a, P = 0.025). In contrast, no local control difference according to IRAK2 expression status was observed in non-irradiated patients (Fig. 4b, P = 0.54). For patients treated with RT, Cox proportional hazard regression confirmed this observation at univariate (HR, 0.301; 95% CI, 0.098–0.923; P = 0.036; Fig. 4c) and multivariate analyses (HR, 0.243; 95% CI, 0.071–0.838; P = 0.025; Fig. 4d). In multivariable analysis, we adjusted age, gender, pathological stage, radiotherapy dose, chemotherapy, the status of surgical margin, and expression level of IRAK2 (Table 3). We found that IRAK2 expression (HR = 0.26, 95% CI = 0.10–0.71, P = 0.008) was statistically significantly associated with local recurrence (P = 0.008), while chemotherapy status was not (P = 0.09). However, the outcome benefit on local control did not translate into cancer cause-specific survival (Fig. 4e, f) and overall survival (data not shown) in both the RT and no-RT groups. These results suggest that high IRAK2 expression correlates with high local control in OSCC patients treated with RT, but not in those without RT.

Table 2

Clinicopathological features of 172 oral cancer patients according to the expression level of IRAK2

	Low-expressed (n = 87)		High-expressed (n = 85)		P value
Age
Age ≦ 50	32	36.8%	32	37.6%	0.91
Age > 50	55	63.2%	53	62.4%
Gender
Male	76	87.4%	80	94.1%	0.13
Female	11	12.6%	5	5.9%
Pathology grade
G1	8	9.2%	4	4.7%	0.51
G2	73	83.9%	75	88.2%
G3	6	6.9%	6	7.1%
pT
pT1	39	44.8%	20	23.5%	0.01
pT2	32	36.8%	34	40.0%
pT3	6	6.9%	10	11.8%
pT4	10	11.5%	21	24.7%
pN
pN0	73	83.9%	61	71.8%	0.14
pN1	7	8.0%	14	16.5%
pN2	7	8.0%	10	11.8%
Pathological stage
I	36	41.4%	19	22.4%	0.02
II	26	29.9%	24	28.2%
III	11	12.6%	15	17.6%
IVA-IVB	14	16.1%	27	31.8%
RT
No	44	50.6%	35	41.2%	0.22
Yes	43	49.4%	50	58.8%
CT
No	51	58.6%	43	50.6%	0.29
Yes	36	41.4%	42	49.4%
CCRT
No	64	73.6%	53	62.4%	0.12
Yes	23	26.4%	32	37.6%
Treatment
OP alone	40	46.0%	30	35.3%	0.38
OP + RT	20	23.0%	18	21.2%
OP + RT/CT	4	4.6%	5	5.9%
OP + CCRT	23	26.4%	32	37.6%
Margin status
Margin < 1mm	11	12.6%	20	23.5%	0.06
Margin ≧ 1mm	76	87.4%	65	76.5%
Bone invasion
Negative	81	93.1%	67	78.8%	0.01
Positive	6	6.9%	18	21.2%
Extracapsular spread of lymph node
Negative	85	97.7%	80	94.1%	0.28
Positive	2	2.3%	5	5.9%
Lymphatic permeation
Negative	82	94.3%	74	87.1%	0.10
Positive	5	5.7%	11	12.9%
Vascular permeation
Negative	86	98.9%	80	94.1%	0.12
Positive	1	1.1%	5	5.9%
Perineural invasion
Negative	73	83.9%	70	82.4%	0.79
Positive	14	16.1%	15	17.6%
Submandibular gland invasion
Negative	87	100.0%	83	97.6%	0.24
Positive	0	.0%	2	2.4%
Skin invasion
Negative	83	95.4%	82	96.5%	0.72
Positive	4	4.6%	3	3.5%
Abbreviation: RT, radiotherapy; CT, chemotherapy; CCRT, concurrent chemoradiotherapy; OP, surgical operation.

Table 3

Univariate (crude) and multivariate (adjusted) logistic regression analysis
	Crude estimate					Adjusted estimate
	Coefficient	HR	95% CI		p-value	Coefficient	HR	95% CI		p-value
IRAK2 (Low, ref)	-0.90	0.41	0.16	1.02	0.056	-1.33	0.26	0.10	0.71	0.008
Age (Continuous)	-0.01	0.99	0.96	1.02	0.52	-0.02	0.98	0.94	1.02	0.33
Gender (Male, ref)	-0.33	0.72	0.17	3.06	0.66	-0.29	0.75	0.17	3.28	0.70
Grade 1 (ref)
Grade 2	0.52	1.69	0.23	12.51	0.61	-0.17	0.85	0.10	7.04	0.88
Grade 3	-12.59	0.00	0.00	.	0.98	-12.56	0.00	0.00	.	0.98
pStage I (ref)
pStage II	-0.31	0.73	0.26	2.06	0.55	0.29	1.34	0.45	3.95	0.60
pStage III	0.84	2.31	0.86	6.22	0.10	1.26	3.52	1.00	12.41	0.05
pStage IV	0.51	1.66	0.36	7.71	0.51	0.88	2.40	0.39	14.81	0.35
OP (ref)
OP with adjuvant RT	1.29	3.64	1.24	10.67	0.018	1.42	4.16	1.32	13.07	0.015
OP with adjuvant CT	1.54	4.66	0.90	24.03	0.07	1.60	4.94	0.78	31.33	0.09
OP with adjuvant CCRT	1.04	2.84	0.90	8.94	0.08	0.09	1.09	0.26	4.52	0.91
Factor score (Continuous)	0.76	2.14	1.24	3.71	0.007	0.70	2.02	1.00	4.06	0.05
* HR: hazard ratio; OP, surgery; RT, radiotherapy; CT, chemotherapy; CCRT, concurrent chemoradiotherapy; Factor score: pathology risk factor and surgical margin < 1mm

IRAK2, an essential mediator of the IL-1R/ TLR signaling, has been recognized to regulate immune response[4]. Recently, IRAK2 has been implicated in playing several roles in human cancers. For example, IRAK2 mediates phosphorylation of Smurf1, which triggers ER stress-mediated apoptosis in colorectal cancer cells[11]. IRAK2 has also been described as a prognostic factor for NSCLC patient survival[12]. Of note, our previous study was the first to discover that IRAK2 may regulate OSCC radiosensitivity via modulating caspase 8/3-mediated apoptosis[7]. Recently, a member of the IRAK family, IRAK1, was also implicated in radiation response[13, 14]. However, IRAK1 was shown to promote radioresistance, and when targeted, it synergizes with inhibitors of PIN1 to effectively sensitize RT in zebrafish models[13]. Another previous study showed that the SNP rs779901 T allele genotype in IRAK2 was associated with increased expression of IRAK2 mRNA, which is significantly associated with better OS in NSCLC patients[12]. No previous study has reported the effects of IRAK2 in response to RT, which may cause it to become a potential candidate as a significant clinical predictor for irradiated cancer patients.

It is well known that RT modulates the immunomodulatory response[15–17], and the synergy between RT and the immune system is currently receiving significant attention[18–20]. It highlights the need that immune-related biomarkers in radiation-based treatment to predict clinical responses to RT. The present study systematically delineated profiles of radiation-induced gene changes. An OML1 cell lines transcriptomic model was characterized, and 25 upregulated genes were identified in IR-treated cells. Functional analysis, including GO terms[21], indicated that the essential enriched pathways were various types of stress-related and immune system functions. In the present study, IRAK2, a critical factor in the IL-1/TLR-mediated signaling, showed significantly higher expression in irradiated cells; this finding is consistent with our previous laboratory results[7]. Based on functional analysis results, IRAK2 was closely associated with immune response and demonstrates a significant role in the cellular response to IR. Therefore, it may be an effective biomarker for predicting radiation response in irradiated OSCC patients.

The clinical value of targeting IRAK gene family members, specifically IRAK1 and IRAK4, has been elucidated in several cancer types[22–24]. For example, IRAK1 was upregulated in hepatocellular carcinoma (HCC) tissue. High IRAK1 expression was associated with large tumor size, metastasis, advanced T status, and poorer OS in HCC patients[22]. Similarly, activated IRAK4 (p-IRAK4) is associated with poor prognosis in colorectal cancer (CRC) patients with stage IIb–IV more notably than that of those patients with stage I-II[24]. Our previous study preliminarily observed that high-expressed IRAK2 may be associated with better local control in early pathological stage OSCC patients treated with RT[7]. Furthermore, the present study confirmed this finding and showed that higher IRAK2 expression correlated with advanced clinical features, including pT3-4 status, overall pathological III-IVB stage, and positive bone invasion. Though molecular findings support the possible prognostic value of IRAK2. However, we found that the observed local control benefit of high IRAK2 expression did not translate into overall survival. It may have some reasons. First, the patient number is limited. This factor may mask the prognostic values of IRAK2. Second, this was a retrospective study, a number of factors in terms of patient and tumor characteristics could not be controlled. Third, the records of 172 OSCC patients were included all stages. The complicated treatment modalities would influence the analysis. For example, most treatment failure of oral cancer post treatment is local recurrence. Surgery is still effective strategy in salvage setting, especially in early oral cancer. These kinds of groups would decrease the prognostic value of survival by IRAK2. However, our study demonstrated high IRAK2 expression predicted higher post-irradiation local control. IRAK2 was associated with radiation response. These findings support IRAK2 as a potential radiation biomarker. Further investigation is suggested.

For treatment response, a high expression of p-IRAK4 has been reported to correlate with a poor chemotherapy response[24]. The poor prognosis was associated with higher clinical stages, larger tumor size, and treatment modalities[25–27]. Our results showed that irradiated patients with high IRAK2 protein expression on IHC stains demonstrated high local control. Few studies have investigated the clinical significance of IRAK2 in the irradiated OSCC patient population. Our results supported the possibility that IRAK2 expression is potentially predictive of the local control rate for irradiated OSCC patients.

Evidence from various pre-clinical and clinical studies showed that immune-related genetic factors are potential biomarkers in cancer diseases[15, 28, 29]. Moreover, it has been demonstrated that a tumor and its microenvironment could induce innate and adaptive immune responses under radiation therapy. These responses may exert roles with either pro- or antitumoral properties[30]. Other IRAK family members, such as IRAK1 and IRAK4, have been shown clinical values for certain cancers. Till now, no other research evidence suggests that IRAK family members play a vital role in tumor response to radiation therapy. Our study is the first report that shows IRAK2 is associated with RT response in cancer patients. However, detailed clinical significance and mechanisms for IRAK2 to trigger post-IR cellular response have not yet been fully resolved. Thus, additional studies are required to further evaluate the mechanisms involved in the relationship between IRAK2 and irradiation response, and prospective studies are encouraged and necessary to support our results.

As a result, the present study expanded prior RNA sequencing-based data to functional biological analysis, IRAK2 plays a crucial role in the radiation-induced response. Clinically, patients with a high expression level of IRAK2 demonstrated more advanced disease features but predicted higher post-irradiation local control than those with low expression. These findings support IRAK2 as a potential predictive biomarker for RT response in OSCC patients.

OSCC: Oral squamous cell carcinoma; RT: Radiotherapy; IR: Ionizing radiation; OS: Overall survival; IRAK2: Interleukin-1 receptor-associated kinase 2; IL-1R: Interleukin-1 receptor; TLR: Toll-like receptor; VMAT: Volumetric-modulated Arc Therapy; GEO: Gene Expression Omnibus; RPKM: Reads Per Kilobase of transcript per Million reads mapped; GO: Gene ontology; DAVID: Visualization, and Integrated Discovery.

Acknowledgements

Not applicable.

Authors’ contributions

C-CY, R-IL, H-YL and F-CH performed the experiments. L-CC, L-WH, C-HC and H-JY collected patient samples. W-YC, C-LC, M-SL and MW. Y.C performed data analysis. MW. Y.C, H-YL and S-KH designed the experiments. C-CY, H-YL and S-KH wrote the manuscript.

Funding

This study was supported by grants from the Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation (DTCRD108-I-17, DTCRD108(2)-I-23, DTCRD109(2)-I-03, DTCRD110-I-06), Buddhist Tzu Chi Medical Foundation (TCMF-A 111-10, TCMF-A 112-04, TCMF-A 108-06) and Ministry of Science and Technology, Taiwan (MOST 108-2314-B-303-001, MOST 109-2314-B-303 -014 -MY2).

Competing interests

The authors declare that they have no competing interests.

Ethics approval and consent to participate

The Institute Review Board (IRB) of the Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, approved the study design and research methods of the present study (registered number: B10704009 and B10604021) prospectively. The IRB waived informed consent and conducted regular ethics visits per protocol. No ethic-harming events were identified during the whole study process of data collection, analysis, and interpretation.

Availability of data and materials

All data supporting the conclusions of this article is available upon request.

Consent for publication

Not applicable.

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No competing interests reported.

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IRAK2, an immune and radiation-response gene, correlates with advanced disease features but predicts high post-irradiation local control in resected oral cancer patients

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Abstract

Figures

Background

Methods

Clinical specimens

Cell culture and radiation treatment

Differential gene expression profile in response to radiation

Functional analysis of radiation-responsive gene

Western Blotting

Immunohistochemistry

Research Database of Clinical Outcomes

Statistical Analysis

Results

Investigating irradiation-response genes in irradiated OML1 oral cancer cells through transcriptome analysis

Gene ontology analysis shows enriched biological processes of post-irradiation upregulated genes

IRAK2 expression predicts local control for patients who received radiotherapy

Discussion

Conclusions

Abbreviations

Declarations

References

Additional Declarations

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