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Genetic association and functional implications of TLR4 rs1927914 polymorphism on colon cancer risk

BMC Cancer volume 24, Article number: 858 (2024) Cite this article

Abstract

Background

Colon cancer remains a major health concern worldwide, with genetic factors playing a crucial role in its development. Toll-like receptors (TLRs) has been implicated in various cancers, but their role in colon cancer is not well understood. This study aims to identify functional polymorphisms in the promoter and 3′UTR regions of TLRs and evaluate their association with colon cancer susceptibility.

Methods

We conducted a case-control study involving 410 colon cancer patients and 410 healthy controls from the Chinese population. Genotyping of polymorphisms in TLR3, TLR4, TLR5 and TLR7 was performed using PCR-RFLP and TaqMan MGB probes. Using logistic regression analysis, we evaluated the association of TLRs polymorphisms and the susceptibility to colon cancer. To understand the biological implications of the TLR4 rs1927914 polymorphism, we conducted functional assays, including luciferase reporter assay and electrophoretic mobility shift assay (EMSA).

Results

Our results demonstrated that the G-allele of the TLR4 rs1927914 polymorphism is significantly associated with a decreased risk of colon cancer (OR = 0.68, 95%CI = 0.50–0.91). Stratified analysis showed that TLR4 rs1927914 AG or GG genotype contributed to a decreased risk of colon cancer among younger individuals (OR = 0.52, 95%CI = 0.34–0.81), males (OR = 0.58, 95%CI = 0.38–0.87), non-smokers (OR = 0.58, 95%CI = 0.41–0.83) and non-drinker with OR (95%CI) of 0.66 (0.46–0.93). Functional assays demonstrated that in HCT116 and LOVO colon cancer cells, the luciferase activity driven by the TLR4 promoter with the rs1927914A allele was 5.43 and 2.07 times higher, respectively, compared to that driven by the promoter containing the rs1927914G allele. Electrophoretic mobility shift assay (EMSA) results indicated that the rs1927914G allele enhanced transcription factor binding. Using the transcription factor prediction tool, we found that the G allele facilitates binding of the repressive transcription factor Oct1, while the A allele does not.

Conclusion

The TLR4 rs1927914 polymorphism influence the susceptibility to colon cancer, with the G allele offering a protective effect through modulation of gene expression. These insights enhance our understanding of the genetic determinants of colon cancer risk and highlight TLR4 as a promising target for cancer prevention strategies.

Peer Review reports

Introduction

Inflammation, a crucial component of innate immunity, is now recognized as a hallmark of cancer development and progression [1]. Chronic inflammation significantly impacts various cancers, including colon cancer [2]. The inflammatory response promotes carcinogenesis through multiple mechanisms, such as the anti-apoptotic effect of nuclear factor-κB (NF-κB), DNA oxidative damage, and alterations to the tumor environment [3,4,5]. Toll like receptors (TLRs) are important innate immune molecules that belong to the family of pattern recognition receptors (PRRs) [6]. TLRs induce the release of various cytokines and inflammatory factors by recognizing and binding to pathogen-related molecular patterns (PAMP), thereby participating in the immune inflammatory response [7]. Specifically, TLR5 on intestinal epithelial cells regulates the composition of intestinal flora and helps prevent inflammation-related diseases [8]. TLRs are expressed not only on immune cells, but also on human tumor cells [9, 10].

According to GLOBOCAN 2022, colorectal cancer is the third most common cancer and the second leading cause of cancer-related deaths worldwide [11]. Environmental factors and lifestyle choices, such as smoking and drinking, are associated with an increased risk of colon cancer [12, 13]. However, it remains unclear why colon cancer develops in some individuals but not others, even exposed to the same risk factors. Studies suggest that genetic variants in cancer-related genes could influence an individual’s susceptibility to colon cancer [14,15,16].

Given the significant role of TLRs in cancer development, we conducted a case-control study to investigate the potential functional single nucleotide polymorphisms (SNPs) in TLRs and their contribution to colon cancer development.

Materials and methods

Study population

This study includes 410 colon cancer patients and 410 healthy controls. Cases were collected from Jan 2008 to Dec 2016 at Tangshan Renmin Hospital and Gongren Hospital of North China University of Science and Technology (NCST) in China with histopathological confirmation. Cancer-free healthy controls were collected from Tangshan area. All participants were genetically unrelated Han Chinese. Informed consent was provided by every individual. Participant provides 2 ml venous blood sample for DNA extraction. This research was supported by the Institutional Review Board of North China University of Science and Technology.

The selection of TLRs genetic variants

Based on the HCB data from NCBI database, we selected variants with a minor allele frequency (MAF) greater than 0.05 in the promoter and 3’ untranslated regions in TLR3, TLR4, TLR5 and TLR7 genes. We used the web-based transcription factor prediction tools (Alibaba2.1 and JASPAR) to predict transcription factor binding sites. Additionally, the SNPinfo web server and mirSNP were applicated to predict microRNA binding sites.

Determination of TLRs genotypes

In this study, genotyping was performed on DNA extracted from peripheral blood samples of colon cancer patients and healthy controls to identify germline polymorphisms associated with colon cancer risk. It is critical to emphasize that the DNA used for genotyping was derived from peripheral blood rather than tumor tissue, thus primarily reflecting the germline genetic status rather than the somatic variations that might be present within tumor cells.

The genotypes for TLR4 rs1927914 and rs7869402 polymorphisms were determined by Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. The primers were synthesized by SinoGenoMax (China, Beijing). The primer pairs used were rs1927914F/ rs1927914R (5′-TAG CAT GAG AAA TGA GGA AGT AAG GG-3′/ 5′-GAG CTA TGA TGA GGA TTG AAA ATG TGG-3′) and rs7869402F/ rs7869402R (5′-TGG GAT CCC TCC CCT GTA GC-3′/ 5′-AGG AGC ATT GCC CAA CAG G-3′). PCR was performed in a 6 µl reaction mixtures containing 20ng genomic DNA, 0.1µM each primer and Taq PCR StarMix (GenStar, China). The PCR procedure involved an initial denaturation step at 94 °C for 5 m, followed by 30 cycles of denaturation at 94 °C for 20s, annealing at 59 °C for 30s, and extension at 72 °C for 35s, with a final extension at 72 °C for 5 m. The PCR products for amplifying TLR4 rs1927914 (524 bp) and rs7869402 (102 bp) were digested by NSi I and Alu I (NEB, Ipswich, USA), respectively, and the resulting fragments were analyzed by agarose-gel electrophoresis.

To genotype other genetic variants, TaqMan MGB probe-based technique were used. TaqMan SNP genotyping assays (C_27310258, C_8812434 and C_2259573) (Thermo Fisher Scientific, Waltham, USA) were used to genotype TLR3 rs5743303, TLR5 rs1640816 and TLR7 rs3853839, respectively.

To ensure the accuracy and reliability of our genotyping results from PCR-RFLP and TaqMan assays, we implemented a rigorous validation protocol. We randomly selected 10% of the samples from each identified genotype for duplicate testing to assess the consistency and reproducibility of our data. The results from these repeated assays demonstrated perfect consistency, with no discrepancies between the initial and follow-up tests. Additionally, the genotyping results were further confirmed by direct sequencing.

The construction of TLR4 luciferase reporter vectors

To confirm the role of TLR4 promoter variants on transcriptional activity, we prepared allelic reporter constructs containing 1832 bp fragment of the TLR4 promoter region, spanning from − 1762 bp to 70 bp. The primers used were 5’-GGG GTA CCC CGG ATT GGA AGT GCT TGG AG-3’ and 5’-CTA GCT AGC TAG AAG AAG AAA ACG CCT GC-3’, which included Kpn I and Nhe I (NEB, Ipswich, USA) cloning sites (underlined sequences). The PCR product was digested by Kpn I and Nhe I and then cloned into pGL3-basic luciferase reporter vector (Promega, Madison, USA).

We designed this construct as pGL3-rs1927914A based on the sequence results. Subsequently, we obtained pGL3-rs1927914G plasmids by site-specific mutagenesis using the pGL3-rs1927914 A vector as template. All constructs were verified by direct sequencing.

Cell culture and transfection for luciferase reporter assay

Colon cancer cell lines (HCT116 and LOVO) were cultured in DMEM medium supplemented with 10% FBS. For each well, 3 × 105 cells were seeded in a 24-well plate and cultured to 80% confluence. The cells were then co-transfected with different pGL3-based constructs and pRL-SV40 using Lipofectamine™ 2000 reagent (Invitrogen, Carlsbad, USA). After transfection, luciferase activities were measured, and the ratio of firefly to Renilla luciferase activities was calculated to determine relative promoter activity. Independent experiments were performed three times.

Electrophoretic mobility shift assay (EMSA)

Biotin-labeled oligonucleotide probes specific for the rs1927914A (5’-TCT AGG ACT TAG CAT ACA AAT ATT CCT GTT-3’) and the rs1927914G (5’-TCT AGG ACT TAG CAT GCA AA TAT TCC TGT T-3’) variants were synthesized (Sangon Biotech; Shanghai, China). DNA binding ability was assessed using a LightShift™ Chemiluminescent EMSA kit (ThermoFisher Scientific, Waltham, USA). Competitors, identical to the labeled probes but without biotin conjugation, were used to confirm the specificity of the DNA-protein interactions. Nuclear proteins were extracted from HCT116 cells using NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, Waltham, USA). Nuclear extracts were incubated with 20fmol labeled oligonucleotide probes for 15 min. For competition experiment, 4pmol of unlabeled oligonucleotide competitors were added prior to incubation with the labeled probes. The binding reactions were then subjected to electrophoresis in a 6.5% polyacrylamide gel. The reactions were then transferred to positively charged nylon membrane and examined for chemiluminescence.

Statistical analysis

The differences in basic demographic information between the healthy population and colon cancer patients were examined using the χ2 test. Hardy-Weinberg Equilibrium of all tested SNPs among controls were also estimated by the χ2 test. The associations of genetic variants in TLR genes with colon cancer risk were evaluated by calculating the odds ratios (OR) with 95% confidence intervals (CI), adjusting for possible confounding factors.

Current smokers were defined as those who had smoked up to one year prior to diagnosis (for colon cancer patients) or up to the date of the interview (for controls). Smoking dose was indicated by pack-years, and smokers were categorized into light and heavy smokers based on the median of pack-years value in controls. Differences in luciferase activity were determined by the t test. Data analyses were performed using SPSS 23.0 (SPSS, Chicago, USA).

Results

Participant information

The basic information for all participants were summarized in Table 1. The median age for both groups was 60 years old. There were no significant differences in the distribution of gender and age between patients and health controls. The distributions of smoking status and drinking status did not differ significantly between the two groups (P > 0.05).

Table 1 Distributions of selected characteristics of patients with colon cancer cases and controls
Full size table

Association of TLRs polymorphisms with colon cancer risk

Bioinformatics analysis predicted that 5 SNPs in the promoter and 3’UTR regions of TLRs could affect transcription factor or microRNA binding. For SNPs with a limited number of complete mutations, we combined genotypes with at least one mutant allele for further analysis. The genotype frequencies for each SNP and their association with the risk of colon cancer were presented in Table 2.

Table 2 Genotype frequencies of toll-like receptor family gene and their association with Colon cancer
Full size table

Using a multivariate logistic regression model, we found that the genotype frequencies of TLR4 rs1927914 among cases were significantly different from those among controls. The presence of G-allele in TLR4 rs1927914 was associated with a decreased colon cancer risk (OR = 0.68, 95%CI = 0.50–0.91). This multivariate logistic regression model passed the significance test, with Hosmer-Lemeshow test showing a P value greater than 0.05 and a predicted accuracy exceeding 50%. Cox-Snell’s R2 and Nagelkerke’s R2 were 0.16 and 0.22, respectively. Additionally, the Omnibus test of model coefficients indicated that the model is effective and has good fit (P < 0.05).

Our data didn’t show any association of other TLRs SNPs with the susceptibility to colon cancer.

Stratification analysis of the TLRs variants and colon cancer risk

The stratified analysis results were presented in Table 3. The TLR4 rs1927914G allele containing genotype is associated with a reduced risk of colon cancer in males (OR = 0.58, 95%CI = 0.38–0.87) when stratified by gender. When stratified by age, individuals with at least one rs1927914G allele had a lower risk of colon cancer in younger participants (OR = 0.52, 95%CI = 0.34–0.81), but not in elder participants (OR = 0.87, 95%CI = 0.58–1.31). In analyses stratified by smoking status or drinking status, the TLR4 rs1927914G containing genotype was found to be a protective factor among non-smokers (OR = 0.58, 95%CI = 0.41–0.83) and among non-drinkers (OR = 0.66, 95%CI = 0.46–0.93), but not among smokers or drinkers.

Table 3 Stratified analysis between genotypes of TLR4 rs1927914 and colon cancer risk
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Effect of TLR4 rs1927914 polymorphism on transcriptional activity

We conducted a luciferase reporter assay to investigate the effect of the TLR4 rs1927914 polymorphism on transcriptional activity. Constructor pGL3-rs1927914A or pGL3-rs1927914G were co-transfected with pRLSV40 control plasmid in HCT116 and LOVO cells. As showed in Fig. 1A, the results showed that he luciferase activity driven by rs1927914A-containing TLR4 promote was 5.43 times higher in HCT116 cells and 2.07 times higher in LOVO cells compared to the rs1927914G-containing TLR4 promoter. These results indicated that the TLR4 rs1927914G allele was associated with a strikingly lower promoter activity, suggesting a functional impact of this variant on TLR4 expression.

Fig. 1
figure 1

The functional analysis of TLR4 rs1927914 polymorphism. (A) Luciferase expression of two constructers (pGL3-rs1927914G and pGL3-rs1927914A) in HCT116 and LOVO cells co-transfected with pRL-SV40 to standardize transfection efficiency. Fold increase was measured by setting the activity of the empty pGL3-Basic vector as 1. *P < 0.05 **P < 0.01 compared with each of the construct counterparts. (B) Electrophoretic mobility shift assays with biotin-labeled oligonucleotide probes containing TLR4 rs1927914 A or G allele. Nuclear extracts were incubated with 5’-Biotin-TCTAGGACTTAGCATACAAATATTCCTGTT-3’ (A probe, lanes 1–3) or 5’-Biotin-TCTAGGACTTAGCATGCAAATATTCCTGTT (G probe, lanes 4–6). Lanes 1 and 4 show the gel mobilities of the biotin-labeled probes without nuclear extracts; lanes 2 and 5 show the mobilities of the biotin-labeled probes with nuclear extracts in the absence of unlabeled probes. The binding specificity was confirmed by competing the biotin-labeled A or G probe with a 100-fold molar excess of unlabeled A (lane 3) or G probe (lane 6). (C) Transcription factor prediction using JASPAR and AliBaba showed that rs1927914 was consistent with the binding sequence of Oct-1

Full size image

Given that allele-specific activity of genetic variants in regulatory regions can influence the binding affinity of transcription factors (TFs), we further conducted an EMSA to investigate if different TLR4 rs1927914 alleles effect the binding activity of transcriptional factor (Fig. 1B). Nuclear extracts prepared from HCT116 cells were incubated with biotin-labeled oligonucleotides for the rs1927914 locus containing either the A or G allele. As shown in the lane 5, shifted bands were observed when the nuclear extract was incubated with the biotin-labeled probe containing the rs1927914G allele, indicating a protein-DNA interaction. In contrast, no shifted bands were observed with the biotin-labeled probe containing the rs1927914A allele (lane 2), suggesting a lack of binding. To confirm the specificity of the binding complex, a 100-fold molar excess of unlabeled oligonucleotide probe was added to the reaction. This competition assay inhibited the formation of the binding complex, as seen in lane 6, further confirming the specificity of the interaction between the nuclear extract and the rs1927914G allele probe. These results demonstrated that the rs1927914G allele had a stronger binding affinity to nuclear protein compared to the A allele. This suggests that TLR4 rs1927914G allele likely increases the binding of transcription factor, subsequently leading to a significant decrease in TLR4 promoter activity.

Our electrophoretic mobility shift assay (EMSA) findings prompted us to further explore the impact of the TLR4 rs1927914 genetic variation on transcription factor binding using JASPAR and Alibaba transcription factor prediction website. The resluts indicated that the TLR4 promoter sequence with the rs1927914G allele can bind to Oct1, whereas that with the rs1927914 A allele cannot (Fig. 1C).

These results suggest that the rs1927914G allele enhances repressive transcription factor Oct1 binding to the TLR4 promoter, leading to decreased transcriptional activity of the TLR4 gene. This mechanism likely explains the reduced TLR4 promoter activity observed with the G allele in our luciferase reporter assays. Therefore, the presence of the rs1927914G allele in the TLR4 promoter region may result in lower TLR4 expression levels due to the increased binding of the repressive transcription factor Oct1.

Discussion

TLRs are crucial components of the inflammatory response, impacting the innate immune response. In recent years, numerous studies have elucidated the roles and molecular mechanisms of TLRs in the development of various cancers [17, 18]. Cancer susceptibility is influenced by SNPs in these TLRs, which can serve as potential biomarker for assessing cancer risk. Genetic variants in TLRs may alter ligand binding capability and subsequently modulate cancer risk [19, 20]. Studies has indicated that the effect of TLR4 SNPs on susceptibility to various cancer types is through the disruption of TLR4 signaling [21].

Our study investigated the association between specific TLR polymorphisms and colon cancer risk. Firstly, we found no significant association between colon cancer risk and the TLR3 rs5743303, TLR5 rs1640816, and TLR7 rs3853839 polymorphisms. These variants are rarely reported in relation to various cancers. Notably, TLR7 rs3853839 has been identified in one study as a predictive marker for cetuximab-based chemotherapy in colon cancer patients [22], highlighting its potential clinical relevance in treatment response. For TLR4 rs7869402 polymorphism, it has been reported to reduce the susceptibility to non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), and gastric cancer [23,24,25], suggesting a protective role in these cancers. These findings imply that the rs7869402 variant may contribute to lower incidence rates of these cancer types. However, our data did not show any significant influence of TLR4 rs7869402 on colon cancer risk, indicating that the protective effect of this polymorphism may be cancer-type specific. Additionally, it is worth mentioning that the TLR4 rs7869402 has been associated with reduced overall survival in ovarian cancer patients [26]. This discrepancy underscores the complexity of genetic influences on cancer susceptibility and prognosis, suggesting that the same genetic variant have different impacts depending on the cancer type and possibly other interacting genetic and environmental factors.

Our study indicated that TLR4 rs1927914 polymorphism is associated a decreased colon cancer risk, consistent with findings in other cancers. Case-control studies have shown that the rs1927914 polymorphism is significantly linked to a lower risk of prostate cancer, hepatocellular carcinoma, esophageal squamous cell carcinoma, and small cell lung cancer [24, 27,28,29]. These findings suggest a protective role for the TLR4 rs1927914 polymorphism across various cancer types. However, some reports indicate no association between TLR4 rs1927914 and gastric cancer [25]. These discrepancies suggest that the impact of the TLR4 rs1927914 polymorphism may be context-dependent, influenced by factors such as tissue-specific expression, environmental interactions. The context-dependent nature of these genetic effects highlights the complexity of genetic influences on cancer susceptibility, which may vary by cancer type and population.

Our functional assays provide further insights into the molecular mechanisms underlying these associations. In colon cancer cells, the luciferase activity driven by the TLR4 promoter containing the rs1927914A allele was significantly higher compared to that driven by the promoter with the G allele. Kutikhin et al. found that high TLR4 expression in cancer tissues promoted tumor cell metastasis and invasion [30]. In addition, studies have shown that TLR4 has the potential to become a marker for disease progression in patients with colon cancer [31], and its low expression is correlated with a better prognosis [32]. This suggests that the A allele enhances TLR4 transcriptional activity, potentially facilitating cancer progression. Conversely, the G allele appears to reduce promoter activity, aligning with its protective role against colon cancer.

Our findings indicate that the rs1927914 G allele enhances the binding affinity of transcription factors, specifically facilitating the binding of the repressive transcription factor Oct1, unlike the A allele. This finding aligns with previous studies demonstrating Oct1’s role in transcriptional repression. The interaction between SMRT and the POU domain of Oct-1 further supports the mechanistic basis of our observations [33]. In gastric cancer cells, Oct1 is recruited to the CDX2 promoter but loses its ability to activate transcription, highlighting its complex regulatory role [34, 35]. Additionally, Oct-1 inhibits Slc7a11 and CRP gene expression by binding to their promoters [36, 37]. These insights suggest that the protective effect of the TLR4 rs1927914G allele in colon cancer may result from enhanced Oct-1 binding, leading to reduced TLR4 expression and decreased cancer risk. The context-dependent effects of Oct-1 in different cancer types underscore the complexity of its role in gene regulation and cancer biology.

This study has significant clinical implications for understanding and potentially managing colon cancer risk, particularly in the context of genetic predisposition. The identification of the TLR4 rs1927914 polymorphism as a factor associated with decreased colon cancer risk provides valuable insights into the genetic underpinnings of this disease. Additionally, understanding the role of TLR4 in colon cancer progression opens new avenues for therapeutic interventions. Given that high TLR4 expression is associated with increased metastasis and invasion of tumor cells [32, 38], targeting TLR4 signaling pathways could be a viable strategy for treatment. The identification of the rs1927914 polymorphism’s impact on TLR4 promoter activity further supports the potential of TLR4 as a therapeutic target. However, further research and clinical trials are warranted to translate these findings into practical applications in the clinical setting.

Despite the strengths of our study, several limitations should be acknowledged. One significant limitation is the potential for detection biases inherent in the PCR-RFLP and TaqMan assays. Although we performed duplicate testing and direct sequencing validation on a subset of samples to mitigate this issue, it is possible that some biases or errors could still affect our results. Future studies should consider using multiple genotyping methods or more advanced technologies to further minimize this risk. Additionaly, the study population was limited to the Chinese population, which may affect the generalizability of our findings to other ethnic groups. Genetic variations and their impacts on disease risk can vary significantly across different populations. Lastly, the sample size, while sufficient to detect significant associations, may limit the power to identify more subtle genetic effects or interactions with environmental factors. Larger studies with increased statistical power are needed to uncover additional risk factors and their interactions.

Conclusions

The TLR4 rs1927914 polymorphism influence the susceptibility to colon cancer, with the G allele offering a protective effect through modulation of gene expression. These insights enhance our understanding of the genetic determinants of colon cancer risk and highlight TLR4 as a promising target for cancer prevention strategies.

Data availability

The datasets used during the current study are available from the corresponding author on reasonable request.

Abbreviations

TLRs:

Toll-like receptors

ORs:

Odds ratios

CIs:

Confidence intervals

SNPs:

Single nucleotide polymorphisms

EMSA:

Electrophoretic mobility shift assay

PRRs:

Pattern recognition receptors

PAMP:

Pathogen-related molecular patterns

References

  1. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–74.

    Article  CAS  PubMed  Google Scholar 

  2. Grivennikov SI, Karin M. Inflammatory cytokines in cancer: tumour necrosis factor and interleukin 6 take the stage. Ann Rheum Dis. 2011;70(Suppl 1):i104–108.

    Article  CAS  PubMed  Google Scholar 

  3. Balkwill F, Mantovani A. Inflammation and cancer: back to Virchow? Lancet. 2001;357(9255):539–45.

    Article  CAS  PubMed  Google Scholar 

  4. Coussens LM, Werb Z. Inflammation and cancer. Nature. 2002;420(6917):860–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Karin M, Greten FR. NF-kappaB: linking inflammation and immunity to cancer development and progression. Nat Rev Immunol. 2005;5(10):749–59.

    Article  CAS  PubMed  Google Scholar 

  6. Hajishengallis G, Lambris JD. Microbial manipulation of receptor crosstalk in innate immunity. Nat Rev Immunol. 2011;11(3):187–200.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Kawai T, Akira S. The role of pattern-recognition receptors in innate immunity: update on toll-like receptors. Nat Immunol. 2010;11(5):373–84.

    Article  CAS  PubMed  Google Scholar 

  8. Chassaing B, Ley RE, Gewirtz AT. Intestinal epithelial cell toll-like receptor 5 regulates the intestinal microbiota to prevent low-grade inflammation and metabolic syndrome in mice. Gastroenterology. 2014;147(6):1363–e13771317.

    Article  CAS  PubMed  Google Scholar 

  9. Furrie E, Macfarlane S, Thomson G, Macfarlane GT, Microbiology, Gut Biology G, Tayside T, Tumour B. Toll-like receptors-2, -3 and – 4 expression patterns on human colon and their regulation by mucosal-associated bacteria. Immunology. 2005;115(4):565–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Huhta H, Helminen O, Kauppila JH, Salo T, Porvari K, Saarnio J, Lehenkari PP, Karttunen TJ. The expression of toll-like receptors in normal human and murine gastrointestinal organs and the Effect of Microbiome and Cancer. J Histochem Cytochem. 2016;64(8):470–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Bray F, Laversanne M, Sung H, Ferlay J, Siegel RL, Soerjomataram I, Jemal A. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024;74(3):229–63.

    Article  PubMed  Google Scholar 

  12. Cheng J, Chen Y, Wang X, Wang J, Yan Z, Gong G, Li G, Li C. Meta-analysis of prospective cohort studies of cigarette smoking and the incidence of colon and rectal cancers. Eur J Cancer Prev. 2015;24(1):6–15.

    Article  PubMed  Google Scholar 

  13. Fedirko V, Tramacere I, Bagnardi V, Rota M, Scotti L, Islami F, Negri E, Straif K, Romieu I, La Vecchia C, et al. Alcohol drinking and colorectal cancer risk: an overall and dose-response meta-analysis of published studies. Ann Oncol. 2011;22(9):1958–72.

    Article  CAS  PubMed  Google Scholar 

  14. Deng Y, Wang L, Huang J, Ding H, Wong MCS. Associations between potential causal factors and colorectal cancer risk: a systematic review and meta-analysis of mendelian randomization studies. J Dig Dis. 2022;23(8–9):435–45.

    Article  PubMed  Google Scholar 

  15. Schmit SL, Edlund CK, Schumacher FR, Gong J, Harrison TA, Huyghe JR, Qu C, Melas M, Van Den Berg DJ, Wang H, et al. Novel common genetic susceptibility loci for Colorectal Cancer. J Natl Cancer Inst. 2019;111(2):146–57.

    Article  PubMed  Google Scholar 

  16. Zeng C, Matsuda K, Jia WH, Chang J, Kweon SS, Xiang YB, Shin A, Jee SH, Kim DH, Zhang B, et al. Identification of susceptibility loci and genes for Colorectal Cancer Risk. Gastroenterology. 2016;150(7):1633–45.

    Article  CAS  PubMed  Google Scholar 

  17. Che F, Yin J, Quan Y, Xie X, Heng X, Du Y, Wang L. TLR4 interaction with LPS in glioma CD133 + cancer stem cells induces cell proliferation, resistance to chemotherapy and evasion from cytotoxic T lymphocyte-induced cytolysis. Oncotarget. 2017;8(32):53495–507.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Farrugia M, Baron B. The Role of Toll-Like receptors in Autoimmune diseases through failure of the self-recognition mechanism. Int J Inflam. 2017;2017:8391230.

    PubMed  PubMed Central  Google Scholar 

  19. Kutikhin AG, Yuzhalin AE. Are toll-like receptor gene polymorphisms associated with prostate cancer? Cancer Manag Res. 2012;4:23–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Yang ZH, Dai Q, Gu YJ, Guo QX, Gong L. Cytokine and chemokine modification by toll-like receptor polymorphisms is associated with nasopharyngeal carcinoma. Cancer Sci. 2012;103(4):653–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Kutikhin AG. Impact of toll-like receptor 4 polymorphisms on risk of cancer. Hum Immunol. 2011;72(2):193–206.

    Article  CAS  PubMed  Google Scholar 

  22. Okazaki S, Stintzing S, Sunakawa Y, Cao S, Zhang W, Yang D, Ning Y, Matsusaka S, Berger MD, Miyamoto Y, et al. Predictive value of TLR7 polymorphism for cetuximab-based chemotherapy in patients with metastatic colorectal cancer. Int J Cancer. 2017;141(6):1222–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Wu H, Gao H, Li A, Xie Y, Jia Z, Yang Z, Zhang H, Zhang Z, Zhang X. Impact of genetic variation in TLR4 3’UTR on NSCLC Genetic susceptibility. J Oncol. 2020;2020:7593143.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Zhang H, Gao H, Li A, Wu H, Xie Y, Zhang Z, Zhang X. TLR4 regulatory region variants reduce the susceptibility of small-cell lung cancer in Chinese population. Eur J Cancer Prev. 2022;31(4):363–8.

    Article  CAS  PubMed  Google Scholar 

  25. Li Z, Gao H, Liu Y, Wu H, Li W, Xing Y, Zhang Z, Zhang X. Genetic variants in the regulation region of TLR4 reduce the gastric cancer susceptibility. Gene. 2021;767:145181.

    Article  CAS  PubMed  Google Scholar 

  26. Liang D, Meyer L, Chang DW, Lin J, Pu X, Ye Y, Gu J, Wu X, Lu K. Genetic variants in MicroRNA biosynthesis pathways and binding sites modify ovarian cancer risk, survival, and treatment response. Cancer Res. 2010;70(23):9765–76.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Chen YC, Giovannucci E, Lazarus R, Kraft P, Ketkar S, Hunter DJ. Sequence variants of toll-like receptor 4 and susceptibility to prostate cancer. Cancer Res. 2005;65(24):11771–8.

    Article  CAS  PubMed  Google Scholar 

  28. Minmin S, Xiaoqian X, Hao C, Baiyong S, Xiaxing D, Junjie X, Xi Z, Jianquan Z, Songyao J. Single nucleotide polymorphisms of toll-like receptor 4 decrease the risk of development of hepatocellular carcinoma. PLoS ONE. 2011;6(4):e19466.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Li J, Wu H, Gao H, Kou R, Xie Y, Zhang Z, Zhang X. TLR4 promoter rs1927914 variant contributes to the susceptibility of esophageal squamous cell carcinoma in the Chinese population. PeerJ. 2021;9:e10754.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Kutikhin AG, Yuzhalin AE, Volkov AN, Zhivotovskiy AS, Brusina EB. Correlation between genetic polymorphisms within IL-1B and TLR4 genes and cancer risk in a Russian population: a case-control study. Tumour Biol. 2014;35(5):4821–30.

    Article  CAS  PubMed  Google Scholar 

  31. Cammarota R, Bertolini V, Pennesi G, Bucci EO, Gottardi O, Garlanda C, Laghi L, Barberis MC, Sessa F, Noonan DM, et al. The tumor microenvironment of colorectal cancer: stromal TLR-4 expression as a potential prognostic marker. J Transl Med. 2010;8:112.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Wang EL, Qian ZR, Nakasono M, Tanahashi T, Yoshimoto K, Bando Y, Kudo E, Shimada M, Sano T. High expression of toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer. Br J Cancer. 2010;102(5):908–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Kakizawa T, Miyamoto T, Ichikawa K, Takeda T, Suzuki S, Mori J, Kumagai M, Yamashita K, Hashizume K. Silencing mediator for retinoid and thyroid hormone receptors interacts with octamer transcription factor-1 and acts as a transcriptional repressor. J Biol Chem. 2001;276(13):9720–5.

    Article  CAS  PubMed  Google Scholar 

  34. Qian J, Kong X, Deng N, Tan P, Chen H, Wang J, Li Z, Hu Y, Zou W, Xu J, et al. OCT1 is a determinant of synbindin-related ERK signalling with independent prognostic significance in gastric cancer. Gut. 2015;64(1):37–48.

    Article  CAS  PubMed  Google Scholar 

  35. Almeida R, Almeida J, Shoshkes M, Mendes N, Mesquita P, Silva E, Van Seuningen I, Reis CA, Santos-Silva F, David L. OCT-1 is over-expressed in intestinal metaplasia and intestinal gastric carcinomas and binds to, but does not transactivate, CDX2 in gastric cells. J Pathol. 2005;207(4):396–401.

    Article  CAS  PubMed  Google Scholar 

  36. Lin X, Yang H, Zhang H, Zhou L, Guo Z. A novel transcription mechanism activated by ethanol: induction of Slc7a11 gene expression via inhibition of the DNA-binding activity of transcriptional repressor octamer-binding transcription factor 1 (OCT-1). J Biol Chem. 2013;288(21):14815–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Voleti B, Hammond DJ Jr., Thirumalai A, Agrawal A. Oct-1 acts as a transcriptional repressor on the C-reactive protein promoter. Mol Immunol. 2012;52(3–4):242–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Hsu RY, Chan CH, Spicer JD, Rousseau MC, Giannias B, Rousseau S, Ferri LE. LPS-induced TLR4 signaling in human colorectal cancer cells increases beta1 integrin-mediated cell adhesion and liver metastasis. Cancer Res. 2011;71(5):1989–98.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors thank all participants and volunteers in this study. We are grateful to the editors and anonymous reviewers for the comments and valuable input to improve the quality of manuscript.

Funding

This work is supported by Foundation of Key Project of Natural Science Foundation of Hebei province of China (H2017209233 to X. Zhang) and the Foundation for Talents by Tangshan Human Resources and Social Security Bureau (grant numbers A202110007).

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Authors and Affiliations

  1. School of Public Health, North China University of Science and Technology, Tangshan, 063210, China

    Ang Li, Hui Gao, Hongjiao Wu, Yuning Xie, Zhenxian Jia & Xuemei Zhang

  2. College of Life Science, North China University of Science and Technology, Tangshan, 063210, China

    Ang Li & Xuemei Zhang

  3. Hebei Key Laboratory of Occupational Health and Safety for Coal Industry, Tangshan, 063210, China

    Ang Li & Xuemei Zhang

  4. School of Basic Medical Sciences, North China University of Science and Technology, Tangshan, 063210, China

    Zhenbang Yang

  5. Affliated Tangshan Gongren Hospital, North China University of Science and Technology, Tangshan, 063000, China

    Zhi Zhang

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  1. Ang Li
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  2. Hui Gao
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  3. Hongjiao Wu
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  7. Zhi Zhang
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  8. Xuemei Zhang
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Contributions

AL: acquisition, analysis, and interpretation of data; drafting the manuscript; Z.Z., H.G., and H.W collection data and analysis; Y.X., Y.Z., Z.Y. and Z.J. extraction DNA and acquisition and interpretation of data. X.Z. design of the work and analysis and interpretation of data, revision of the article, final approval of the version to be published. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Xuemei Zhang.

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All the study procedures were approved by the Ethics Committee of North China University of Science and Technology (2019021) and written informed consents were obtained from all participants of their own free will.

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Not applicable.

Competing interests

The authors declare no competing interests.

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Li, A., Gao, H., Wu, H. et al. Genetic association and functional implications of TLR4 rs1927914 polymorphism on colon cancer risk. BMC Cancer 24, 858 (2024). https://doi.org/10.1186/s12885-024-12604-z

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  • DOI: https://doi.org/10.1186/s12885-024-12604-z

Keywords

BMC Cancer

ISSN: 1471-2407

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