Introduction
Wilms tumor is a cancerous kidney tumor that originates from embryonic cells. It is the most prevalent kidney tumor among children aged 0–14 years globally, representing 90% of kidney tumors, with an incidence rate of 3.9 per million.1 Most Wilms tumor patients present with isolated nephroblastoma, while 5–7% exhibit bilateral renal involvement, and 10% have multiple lesions within a single kidney. Nephrogenic rests (NRs), which retain the potential to form embryonic tumors, are considered Wilms tumor precursors. These occur after 36 weeks of gestation and persist as embryonic cells or tissues after birth.2 Notably, NRs play a crucial role in Wilms tumor diagnosis. Approximately 40% of unilateral Wilms tumors and over 90% of bilateral Wilms tumors are accompanied by NRs.3
The cumulative death rate from various causes, excluding recurrence, reaches 5.4% 30 years after Wilms tumor diagnosis, increasing significantly to 22.7% by the 50-year mark.4 At the 50-year milestone, subsequent primary neoplasms and heart disease contribute to cumulative mortality rates of 8.2% and 6.3%, respectively.4 The Renal Tumor Study Group of the International Society of Paediatric Oncology system guides treatment decisions for Wilms tumor. Stage I tumors remain confined to the renal capsule, while stage II tumors infiltrate the renal sinuses, perirenal fat, and adjacent organs without a discernible margin but with a well-defined excision margin. Prechemotherapy-induced tumor rupture is diagnosed through imaging, and chemotherapy-induced lymph node changes serve as stage III indicators. Stage IV tumors involve hematogenous metastases or lymph node metastases beyond the abdominal area.5
Genetic syndromes such as Denys-Drash syndrome and WAGR syndrome (Wilms tumor, absence of the iris, genitourinary abnormalities, and development retardation), are linked to a greater likelihood of developing Wilms tumor.6 The Children’s Oncology Group of America program incorporates molecular markers, including blastemal volume and 1q gain, into clinical decision-making.7 Additionally, a study revealed that altered microRNA (miRNA) expression patterns correlate with a poor prognosis in the blast histology of pediatric Wilms tumor patients treated with chemotherapy.8,9
miRNAs, a class of small noncoding RNAs, play a critical role in cancer risk. Case-control studies have shown that polymorphisms in miRNA-encoding genes are associated with malignant tumors.10,11 Recent findings have suggested that miR-27a in serum/plasma or tumor tissue can serve as a diagnostic biomarker and predict the survival of cancer patients.12 Northern blot analysis has confirmed miR-27a expression in mouse kidneys.13 Additionally, a recent study showed that miR-27a-3p expression is associated with the grade of malignancy in Wilms tumor.14 A single-nucleotide polymorphism (SNP) is a genetic variation that involves a change in a single nucleotide at a specific location within the genome. A case-control study highlighted the protective effects of the miR-27a rs895819 T>C polymorphism against cervical cancer.15 Conversely, a meta-analysis linked the miR-27a rs895819 T>C polymorphism to an increased risk of colorectal cancer in the Chinese population.16 However, no reports have explored the relationship between the miR-27a rs895819 T>C polymorphism and Wilms tumor susceptibility. Therefore, this case-control study aimed to elucidate this association.
Materials and methods
Study population
We included 145 patients with Wilms tumors as the case group and 531 cancer-free patients as the control group (Table S1).17–20 Both the case group and the control group were recruited from Guangzhou Women and Children’s Medical Center. The patients in the case group were diagnosed with Wilms tumor at the hospital and did not have any history of other malignant tumors. These two groups were compared based on age and sex, while the case group was categorized according to clinical stage. This study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). This research was approved by the institutional review committee of Guangzhou Women and Children’s Medical Center (Approval No. 202016601). All participants or their legal guardians provided informed written consent.
SNP selection and genotyping
Total genomic DNA was extracted from paraffin-embedded tissues and peripheral blood using the QIAamp DNA FFPE Tissue Kit (Qiagen, Valencia, CA) and the TIANGEN Blood DNA Extraction Kit (TianGen Biotech, Beijing), respectively. The miR-27a rs895819 T>C polymorphism was genotyped using the TaqMan method and the 7900 sequencing detection system (Applied Biosystems, Foster City, CA, USA). Further methodological details are provided in the cited studies.21–24 To ensure genotyping accuracy, 10% of the samples were retested, and the results were consistent with previous results.
Statistical analysis
Various statistical methods were employed based on variable types. The χ2 test was utilized to evaluate potential disparities between patients and controls. Additionally, the χ2 goodness-of-fit test was utilized to evaluate whether the selected SNP deviated from Hardy-Weinberg equilibrium in the control group. Demographic variables and gene frequencies across subjects were analyzed using two-sided χ2 tests. Multivariate logistic regression analysis was performed to calculate adjusted odds ratios (ORs) and 95% confidence intervals (CIs) to examine the relationship between the rs895819 T>C polymorphism and susceptibility to Wilms tumor. All analyses were performed using the SAS statistical package (version 9.4; SAS Institute, USA), with statistical significance set at P < 0.05.
Results
Association of the miR-27a polymorphism with Wilms tumor susceptibility
The miR-27a rs895819 T>C polymorphism was successfully detected in 145 patients and 531 controls. As presented in Table 1, the genotype distribution of the miR-27a rs895819 T>C polymorphism in both the case and control groups adhered to Hardy-Weinberg equilibrium (P = 0.749). No statistically significant differences were observed between the case and control groups (TC vs. TT: adjusted OR = 1.20, 95% CI = 0.82–1.75, P = 0.358; CC vs. TT: adjusted OR = 0.96, 95% CI = 0.44–2.09, P = 0.923; TT vs. TT/TC: adjusted OR = 1.16, 95% CI = 0.80–1.68, P = 0.425; CC vs. TT/TC: adjusted OR = 0.89, 95% CI = 0.42–1.89, P = 0.757; C vs. T: adjusted OR = 1.08, 95% CI = 0.81–1.44, P = 0.610). We failed to establish a significant association between the miR-27a rs895819 T>C polymorphism and Wilms tumor susceptibility.
Table 1Genotype distributions of miR-27a rs895819 T>C polymorphism and Wilms tumor susceptibility
| Genotype | Cases (n = 145) | Controls (n = 531) | Pa | Crude OR (95% CI) | P | Adjusted OR (95% CI)b | Pb |
|---|
| rs895819 (HWE = 0.749) |
| TT | 72 (49.66) | 285 (53.67) | | 1.00 | | 1.00 | |
| TC | 64 (44.14) | 210 (39.55) | | 1.21 (0.82–1.77) | 0.335 | 1.20 (0.82–1.75) | 0.358 |
| CC | 9 (6.21) | 36 (6.78) | | 0.99 (0.46–2.15) | 0.979 | 0.96 (0.44–2.09) | 0.923 |
| Additive | | | 0.608 | 1.09 (0.82–1.47) | 0.552 | 1.08 (0.81–1.45) | 0.604 |
| Dominant | 73 (50.34) | 246 (46.33) | 0.391 | 1.18 (0.81–1.70) | 0.391 | 1.16 (0.80–1.68) | 0.425 |
| TT/TC | 136 (93.79) | 495 (93.22) | | 1.00 | | 1.00 | |
| CC | 9 (6.21) | 36 (6.78) | 0.806 | 0.91 (0.43–1.94) | 0.806 | 0.89 (0.42–1.89) | 0.757 |
| T | 208 (71.72) | 780 (73.45) | | 1.00 | | 1.00 | |
| C | 82 (28.28) | 282 (26.55) | 0.558 | 1.09 (0.82–1.46) | 0.558 | 1.08 (0.81–1.44) | 0.610 |
Stratification analysis
We conducted a stratification analysis to investigate the association between the rs895819 T>C polymorphism and the risk of Wilms tumor while, taking into account age, sex, and clinical stage (Table 2). The results showed that age (≤18, adjusted OR = 1.11, 95% CI = 0.64–1.93, P = 0.703; >18, adjusted OR = 1.21, 95% CI = 0.74–2.00, P = 0.444), sex (female, adjusted OR = 1.09, 95% CI = 0.63–1.91, P = 0.751; male, adjusted OR = 1.21, 95% CI = 0.74–1.98, P = 0.456), and clinical stage (I+II, adjusted OR = 1.29, 95% CI = 0.73–2.29, P = 0.383; III+IV, adjusted OR = 1.19, 95% CI = 0.75–1.89, P = 0.468) were not significantly associated with the miR-27a rs895819 T>C polymorphism in relation to Wilms tumor susceptibility, even after stratification.
Table 2Stratification analysis for the association of miR-27a rs895819 T>C polymorphism with Wilms tumor risk
| Variables | TT
| TC/CC
| Crude OR
| P | Adjusted ORa
| Pa |
|---|
| (Cases/Controls) | (95% CI) | (95% CI) |
|---|
| Age, month |
| ≤18 | 32/120 | 34/113 | 1.13 (0.65–1.95) | 0.665 | 1.11 (0.64–1.93) | 0.703 |
| >18 | 40/165 | 39/133 | 1.21 (0.74–1.99) | 0.453 | 1.21 (0.74–2.00) | 0.444 |
| Gender |
| Females | 34/129 | 30/104 | 1.09 (0.63–1.91) | 0.750 | 1.09 (0.63–1.91) | 0.751 |
| Males | 38/156 | 43/142 | 1.24 (0.76–2.03) | 0.386 | 1.21 (0.74–1.98) | 0.456 |
| Clinical stage |
| I+II | 25/285 | 28/246 | 1.30 (0.74–2.28) | 0.367 | 1.29 (0.73–2.29) | 0.383 |
| III+IV | 41/285 | 42/246 | 1.19 (0.75–1.89) | 0.468 | 1.19 (0.75–1.89) | 0.468 |
Discussion
In this study, we investigated the association between the miR-27a rs895819 T>C polymorphism and Wilms tumor risk. We found no significant difference in the association between miR-27a rs895819 T>C and Wilms tumor risk.
As a class of noncoding RNA molecules, miRNAs play a pivotal role in negatively regulating the expression of multiple protein-coding genes. They also modulate various cancer-related genes at the posttranscriptional level.25 Recently, miRNA SNPs have garnered attention for their potential association with tumor risk. These SNPs have emerged as candidate biomarkers for breast cancer susceptibility.26,27 According to comprehensive Genome-Wide Association Studies, 26 miRNA SNPs were significantly linked to susceptibility to colorectal cancer, other cancers, obesity, and celiac disease.28 These findings suggest that miRNA SNPs could be targeted in Genome-Wide Association Studies to identify potential susceptibility loci.
Multiple studies have provided evidence for the close association between miR-27a, a small RNA molecule, and cancer susceptibility, including but not limited to renal cell carcinoma, gastric cancer, and colorectal cancer.29,30 Notably, miR-27a expression is downregulated in high-risk Wilms tumors.31 Furthermore, research has shown that miR-27a-5p significantly inhibits Wilms tumor development by suppressing the expression of PBOV1.32 Conversely, in renal cancer cells, miR-27a suppresses growth by targeting ERFR.33 Interestingly, miR-27a upregulation in B4GALT3 has been associated with cervical cancer progression and may serve as a potential biomarker for this disease.34 Additionally, proteomic analysis has linked elevated levels of miR-27a to distant metastasis and poor prognosis in colorectal cancer patients.35 These diverse effects of miR-27a across different malignant tumors likely result from a combination of factors.
The miR-27a rs895819 T>C SNP is located on chromosome 19p13.12 and plays a crucial role in regulating gene polymorphisms, tumorigenesis, proliferation, and apoptosis. While some studies have suggested that the miR-27a rs895819 polymorphism increases the risk of gastric cancer, others have found no association with gastric epithelial dysplasia.36 In a case-control study, the miR-27a rs895819 polymorphism was found to be predictive of renal cell carcinoma risk in the Chinese population.37 We did not find any significant association between the miR-27a rs895819 T>C polymorphism and Wilms tumor risk in our study. However, we observed that the risk of disease tends to decrease linearly with the addition of recessive mutated genes, even if no significant difference was detected. The negative results in this study could be attributed to various factors, including limitations in sample size, diversity among subject groups, geographic variations, genetic background, or potential errors in analysis.
This study faces several challenges that must be addressed for more accurate results. First, Wilms tumor is a complex disease involving multiple genes and intricate interactions. Relying solely on gene polymorphism analysis may lead to inaccurate negative results. Second, increasing the sample size in the case group is essential for establishing a robust correlation between variables. Finally, potential biases during data acquisition and analysis must be carefully considered to ensure the accuracy and reliability of our findings.
Conclusions
Our study provides evidence that there is no association between the miR-27a rs895819 T>C polymorphism and Wilms tumor susceptibility. To validate these findings, further investigations should focus on expanding the sample size and exploring additional genetic polymorphisms.
Supporting information
Supplementary material for this article is available at https://doi.org/10.14218/CSP.2024.00024 .
Table S1
Frequency distribution of selected variables for Wilms tumor cases and cancer-free controls.
(DOCX)
Declarations
Ethical statement
This study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). This research was approved by the institutional review committee of Guangzhou Women and Children’s Medical Center (Approval No. 202016601). All participants or their legal guardians provided informed written consent.
Data sharing statement
The requested data can be readily accessed by reaching out to the corresponding author.
Funding
This work was supported by grants from the National Natural Science Foundation of China (No. 82003523), Guangzhou Science and Technology Project (No. SL2024A04J02146, SL2024A03J01616).
Conflict of interest
The authors have no conflicts of interest related to this publication.
Authors’ contributions
Investigation, manuscript writing (SW), investigation, sample and clinical information collection (CD, YH, WF), conception, data collection, and data analysis (RH). All authors have significantly contributed to the study and have given their approval to the final draft of the manuscript.