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Genetic association of long non-coding RNA ANRIL polymorphism with the risk of type 2 diabetes mellitus in the Chinese Han population

Abstract

Background

Type 2 diabetes mellitus (T2DM) is closely associated with both environmental and genetic factors, involving multi-gene inheritance. This study examined the association between the polymorphic locus rs10757278 in long non-coding RNA ANRIL and T2DM.

Methods

Polymerase chain reaction (PCR) was used to detect the rs10757278 polymorphism in the ANRIL gene. RT-qPCR measured ANRIL expression levels, and logistic regression identified independent risk factors for T2DM. Furthermore, the receiver operating characteristic (ROC) curve was constructed to evaluate the clinical diagnostic value of serum ANRIL levels in diagnosing T2DM.

Results

The rs10757278 polymorphism of the ANRIL was associated with the development of T2DM. Specifically, the G allele increases the risk of T2DM, and individuals carrying the GG genotype have a higher risk of developing the disease. Significant differences were found in low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), and glycated hemoglobin (HbA1c) among T2DM patients with different genotypes of ANRIL rs10757278. The relative FPG and HbA1c levels were relatively lower in individuals with the AA genotype and higher in those with the GG genotype. Moreover, serum ANRIL levels in the T2DM group were lower than in the control group. Body mass index (BMI), the rs10757278 locus, and serum ANRIL levels were independent risk factors for the development of T2DM. The ROC curve showed that serum ANRIL levels have significant clinical diagnostic value for the diagnosis of T2DM.

Conclusion

The rs10757278 polymorphism in ANRIL was strongly associated with the genetic predisposition to T2DM.

Introduction

Type 2 diabetes mellitus (T2DM) is a progressive metabolic disorder influenced by both genetic and environmental factors [1]. The global prevalence of this condition has exhibited a significant upward trend [2]. In China, recent data indicate that, as of 2021, approximately 140.9 million individuals were affected by diabetes, with the majority being adults aged 20 to 79 years. T2DM primarily results from insulin resistance in peripheral tissues [3], constituting 90–95% of all diabetes cases [4]. The etiology of T2DM is well-established as a complex interplay between genetic and environmental factors [5]. To date, genome-wide association studies (GWAS) have identified numerous single nucleotide polymorphisms (SNPs) associated with T2DM [6, 7]. Several susceptibility loci have been consistently replicated across diverse ethnic populations, including the Han Chinese [8, 9]. A thorough investigation into the genetic susceptibility of T2DM can provide a critical theoretical foundation for identifying high-risk populations and developing preventive and control strategies.

Long non-coding RNAs (lncRNAs), lacking open reading frames, are incapable of encoding proteins [10]. These molecules perform vital roles in a multitude of biological processes, including cell proliferation and apoptosis [11, 12]. The lncRNA ANRIL (antisense noncoding RNA in the INK4 locus) was first identified in a melanoma patient with a large (403 kb) deletion at the CDKN2A/B locus [13]. Additionally, studies have reported a significant increase in ANRIL expression in patients with progesterone receptor positivity, suggesting its involvement in breast cancer [14]. ANRIL potentially influences cell growth, differentiation, and survival by modulating intracellular signaling pathways. Specifically, it might disrupt intracellular signal transduction, altering cellular metabolism and behavior, thus impacting the onset and progression of various diseases. Moreover, the expression levels of ANRIL exhibit a significant disparity when compared to those in healthy individuals. This alteration in expression levels could be intricately linked to the pathogenesis of coronary atherosclerotic heart disease (CAD). For instance, ANRIL could be a factor in multiple processes, including inflammatory responses and coronary atherosclerosis [15]. Research has shown that a high-glucose (HG) environment can further potentiate the tumor-promoting effects of ANRIL, alter glucose metabolism, and participate in glycolytic reactions and apoptosis [16]. LncRNA ANRIL is markedly upregulated exposed to high glucose [17,18,19]. ANRIL is involved in the emergence of T2DM, and its dysregulation may serve as a critical factor in the pathogenesis of this condition.

The polymorphism of genes is intricately linked to the susceptibility and pathogenesis of various diseases. Extensive data indicate that certain SNPs associated with diseases exert their effects by influencing ANRIL itself [20]. Specifically, the ANRIL rs1333040 polymorphism is correlated with an elevated risk of colorectal cancer (CRC) and a higher tumor-lymph node-metastasis (TNM) stage. The ANRIL rs10757274 polymorphism correlates with well-differentiated tumors in CRC [21]. The ANRIL rs2383208 and rs10811661 polymorphic sites have been validated to be linked to T2DM, with altered expression levels of ANRIL observed in the blood and islets of affected tissues, along with potential enhancer regions [22, 23]. Notably, the rs10757278 locus on chromosome 9p21 exhibits a highly significant correlation with early-onset and familial myocardial infarction and CAD [24]. However, the precise functional role of the ANRIL rs10757278 polymorphism in T2DM is still not well understood.

This study primarily investigated the correlation between the rs10757278 polymorphism in ANRIL and T2DM in the Chinese Han population. Additionally, a comprehensive evaluation of the association between this polymorphism and the individual characteristics of the non-diabetic group was conducted. By employing detailed statistical analyses and in-depth discussions of these associations, we aimed to elucidate the underlying mechanisms of this polymorphism in ANRIL in T2DM, thereby providing novel insights and a foundation for the prevention and treatment of T2DM.

Materials and methods

Study objects

The study recruited 275 Han patients diagnosed with T2DM at The First Hospital of Hebei Medical University, along with 280 Han healthy controls. The diagnosis of T2DM adhered to the criteria established by the World Health Organization [4], which include a fasting blood glucose level of ≥ 7.0 mmol/L, a 2-hour postprandial blood glucose level of ≥ 11.1 mmol/L, or a random blood glucose level of ≥ 11.1 mmol/L. Exclusion criteria comprised: patients with other types of diabetes (e.g., type 1 diabetes), autoimmune disorders, hypertension, coronary heart disease, malignant tumors, endocrine dysfunction, non-Han ethnicity, and incomplete medical records.

This study was approved by the The First Hospital of Hebei Medical University Medical Ethics Committee (ethics number 20220818), and all participants provided written informed consent. All procedures complied with the principles outlined in the Declaration of Helsinki.

Biochemical measurements

Venous blood (6 mL) was collected from both the control group and the T2DM group after a 12-hour fast. A portion of the collected venous blood was placed in an ethylenediaminetetraacetic acid (EDTA) anticoagulation tube for the determination of glycated hemoglobin (HbA1c) using the immunoturbidimetric method, as well as for gene analysis. Another portion was inserted into a plain vacuum tube lacking anticoagulant, followed by centrifugation to separate the serum for biochemical assessment. Additionally, diastolic blood pressure (DBP) and systolic blood pressure (SBP) were measured. The levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and fasting plasma glucose (FPG) were determined using standardized enzymatic methods.

Genotyping of SNPs

Polymerase chain reaction (PCR) was conducted using a 30 ng DNA sample as the template. Two primer pairs were designed using Premier Primer 5 and Oligo 6.22 software to amplify the polymorphic region encoding the target variation. The PCR reaction conditions were as follows: initial denaturation at 95 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s. A final extension was performed at 72 °C for 10 min. The PCR products were then analyzed by agarose gel electrophoresis. The amplified products were sequenced using Sanger sequencing, and allele recognition was conducted using SDS 2.3 software.

Quantitative real-time PCR (RT-qPCR)

Total RNA from ANRIL was extracted and reverse-transcribed into cDNA. RT-qPCR amplification was performed using this cDNA as the template on a Roche Light Cycler® 96. The expression level of ANRIL was quantified by RT-qPCR analysis. ANRIL -PCR primers were designed with Primer Premier 5 software. ANRIL primer sequences were shown as follows: upstream: 5’-AGGGTTCAAGCATCACTGTTAGG-3’; downstream: 5’-GAAACCCCGTCTCTACTGTTACCT-3’. Following amplification, the relative gene expression level was calculated using the 2−ΔΔCt method, with GAPDH serving as the internal reference for ANRIL.

Statistical analysis

In this study, data processing was conducted using SPSS 16.0 statistical software to analyze the differences in age, gender, and clinical parameters associated with T2DM between the T2DM group and the healthy control group. The Hardy-Weinberg equilibrium (HWE) for rs10757278, the allele and genotype frequencies of rs10757278, and the differences in these frequencies between the T2DM and control groups were evaluated. Count data were presented as percentages (%). The chi-square test was used to compare categorical variables. Logistic regression analysis was performed to identify independent risk factors for T2DM. P < 0.05 was considered statistically significant.

Results

The general clinical characteristics of patients with T2DM

The study included a control group (n = 280) and a T2DM group (n = 275), and their demographic and clinical characteristics were analyzed (Table 1). Regarding age, the mean age in the control group was 58.30 ± 11.78 years, and in the T2DM group, it was 58.15 ± 11.67 years, with no significant difference between the groups (P = 0.877). There were no significant differences in gender distribution between the two groups. (P = 1.000). However, significant differences were observed in other indicators. The body mass index (BMI) was significantly higher in the T2DM group compared to the control group (P < 0.001, (25.33 ± 4.04)kg/m2 vs. (23.16 ± 3.94) kg/m2). For lipid profiles, the T2DM group exhibited higher levels of TC ((4.81 ± 1.13) mmol/L vs. (4.61 ± 1.02) mmol/L), TG ((1.78 ± 0.71) mmol/L vs. (1.43 ± 0.55) mmol/L), and LDL-C ((2.90 ± 0.86) mmol/L vs. (2.69 ± 0.74) mmol/L) compared to the control group (P = 0.028, P < 0.001, P = 0.002, respectively), while HDL-C was lower (1.32 ± 0.32 mmol/L vs. 1.44 ± 0.31 mmol/L, P < 0.001). Blood pressure measurements showed that both SBP ((131.68 ± 14.83) mmHg vs. (124.78 ± 12.83) mmHg) and DBP ((81.48 ± 14.91) mmHg vs. (74.73 ± 10.48) mmHg) were significantly higher in the T2DM group compared to the control group (P < 0.001, P < 0.001, respectively). Additionally, blood glucose indicators, including FPG ((7.60 ± 1.45) mmol/L vs. (4.97 ± 0.63) mmol/L) and HbA1c ((7.01 ± 1.44)% vs. (4.95 ± 0.47)%), were significantly elevated in the T2DM group (P < 0.001, P < 0.001, respectively). In summary, significant differences were found in BMI, lipid profiles, blood pressure, and blood glucose levels between T2DM patients and the control group.

Table 1 Demographic and clinical characteristics of the enrolled study subjects

Genotype or allele frequencies

We conducted a further investigation into the genotype and allele distribution of the rs10757278 polymorphism of ANRIL in both T2DM patients and the control group (Table 2). Regarding alleles, in the control group, 56.79% of individuals carried the A allele, while 43.21% carried the G allele. In contrast, in the T2DM group, 47.45% of individuals carried the A allele, and 52.55% carried the G allele (χ² = 9.682, odds ratio (OR) = 1.455, P = 0.002). These findings indicated a significant difference at the allele level between the T2DM group and the control group for the rs10757278 polymorphism of ANRIL. Specifically, the A allele was more prevalent in the control group, whereas the G allele was more common in the T2DM group. This discrepancy suggested that this polymorphism may be linked to the onset and progression of T2DM, with the G allele potentially increasing the risk of developing the disease.

Table 2 Genotype and allele distributions of LncRNA ANRIL rs10757278 polymorphism inT2DM patients and controls

Under the codominant genetic model, in the control group, 91 individuals (32.50%) had the AA genotype, 136 individuals (48.57%) had the AG genotype, and 53 individuals (18.93%) had the GG genotype; in the T2DM group, 63 individuals (22.91%) had the AA genotype, 135 individuals (49.09%) had the AG genotype, and 77 individuals (28.00%) had the GG genotype. The OR for the AG genotype relative to the AA genotype was 1.434, P = 0.077; the OR for the GG genotype relative to the AA genotype was 2.099, P = 0.002. These results indicated that the GG genotype was significantly associated with an increased risk of T2DM compared to the AA genotype. In the T2DM group, the frequency of the GG genotype was notably higher than in the control group, whereas the frequency of the AA genotype was relatively lower. This suggested that individuals carrying the GG genotype were at a higher risk of developing T2DM, and the AG genotype may also be associated with T2DM, although this association does not reach statistical significance.

Under the dominant genetic model, in the control group, 91 individuals (32.50%) had the AA genotype, while 189 individuals (67.50%) had the AG + GG genotype; in the T2DM group, 63 individuals (22.91%) had the AA genotype, and 212 individuals (77.19%) had the AG + GG genotypes (χ² = 6.366, OR = 1.620, P = 0.012). These findings indicated a significant difference in genotype distribution between the T2DM group and the control group under the dominant genetic model. This suggested that individuals with the AG + GG genotypes were at a higher risk of developing T2DM compared to those with the AA genotype.

Under the recessive genetic model, in the control group, 227 individuals (81.07%) had the AA + AG genotype, while 53 individuals (18.93%) had the GG genotype; in the T2DM group, 198 individuals (72.00%) had the AA + AG genotype, and 77 individuals (28.00%) had the GG genotype (χ² = 6.365, OR = 1.666, P = 0.012). These results indicated a significant difference in genotype distribution between the two groups under the recessive genetic model. Specifically, individuals carrying the GG genotype exhibited a higher risk of developing T2DM compared to those with the AA + AG genotype. The population was found to conform to Hardy-Weinberg equilibrium (PHWE = 0.863). Therefore, the rs10757278 polymorphism of ANRIL may be associated with the development of T2DM, with the G allele potentially increasing the risk of the disease. Notably, individuals with the GG genotype have a higher risk of developing T2DM, and the frequency of the AA genotype was relatively lower in the T2DM group.

The effects of different genotypes on blood glucose-related indicators

We further investigated the demographic and clinical characteristics of all subjects carrying different ANRIL rs10757278 genotypes (Table 3). In the control group, statistical analysis revealed no significant differences among individuals with different genotypes in terms of age, sex, BMI, TC, TG, HDL-C, LDL-C, SBP, DBP, FPG, and HbA1c (P > 0.05). In the T2DM group, regarding age, the mean ages for the AA, AG, and GG genotypes were 57.90 ± 12.86 years, 58.45 ± 11.61 years, and 57.81 ± 10.88 years, respectively, with no statistically significant differences among the groups (P = 0.912). In terms of gender distribution, the difference was also not significant (P = 0.725). For BMI, the means for the AA, AG, and GG genotypes were 25.61 ± 4.38 kg/m², 24.95 ± 4.20 kg/m², and 25.78 ± 3.39 kg/m², respectively, with no significant differences (P = 0.293). TC levels were also similar across the three groups (P = 0.205). The differences in TG and HDL-C levels among patients with different genotypes were not significant, with P values of 0.501 and 0.229, respectively. However, LDL-C levels showed a highly significant difference among the genotypes (P < 0.001). SBP and DBP did not differ significantly among patients with different genotypes, with P values of 0.217 and 0.238, respectively. FPG levels in patients with the AA, AG, and GG genotypes were 6.86 ± 1.46 mmol/L, 7.58 ± 1.35 mmol/L, and 8.23 ± 1.34 mmol/L, respectively. HbA1C levels in patients with different genotypes were 6.34 ± 1.36%, 7.01 ± 1.45%, and 7.55 ± 1.28%, respectively. The differences in both FPG and HbA1C levels among patients with different genotypes were highly statistically significant (all P < 0.001). In summary, among T2DM patients with different ANRIL rs10757278 genotypes, no significant differences were observed in age, gender, BMI, TC, TG, HDL-C, SBP, or DBP. However, significant differences were noted in LDL-C, FPG, and HbA1C.

Table 3 Demographic and clinical characteristics of the T2DM patients carrying different ANRIL rs10757278 genotypes

The levels of FPG and HbA1C were different in individuals with different genotypes. In the control group, no significant differences were observed in the effects of various genotypes on relative FPG (Fig. 1A) and HbA1c (Fig. 1B) levels. The data illustrated that, in contrast to the control group, different genotypes had a significant impact on both relative FPG (Fig. 1C) and HbA1c (Fig. 1D) levels in the T2DM group. A discernible pattern emerges: the relative FPG and HbA1C levels were notably lower in individuals with the AA genotype, whereas they were markedly higher in those with the GG genotype. This variation likely stems from genetic factors. Across the different genotypes (AA, AG, GG), the trends in FPG and HbA1C levels were consistent, suggesting a coordinated response to these genetic influences. In summary, the distinct genotypes (AA, AG, GG) exerted a substantial impact on the relative levels of FPG and HbA1C. This insight contributed to a more profound understanding of the genetic mechanisms underlying blood glucose metabolism and offered valuable data for future research and clinical applications.

Fig. 1
figure 1

(A) The FPG level of carriers with different genotypes. (B) The HbA1C level of carriers with different genotypes. * means P < 0.05, ** means P < 0.01, *** means P < 0.001

The expression level of ANRIL decreased in T2DM patients

We conducted a more detailed investigation into the differences in the relative expression levels of ANRIL between the control group and individuals with T2DM (Fig. 2A), as well as among different genotypes (AA vs. AG/GG) (Fig. 2B). This analysis aimed to elucidate the relationship between ANRIL gene expression, T2DM, and genetic variations. The findings revealed that irrespective of T2DM or genotype, the serum ANRIL levels in the T2DM group were significantly lower (Fig. 2A). Within both the control and T2DM groups, individuals with the AA genotype exhibited higher serum ANRIL levels than those with the AG/GG genotype (Fig. 2B). These observations indicated that the ANRIL gene may have a significant role in the development of T2DM. Additionally, the substantial variability in relative ANRIL expression levels observed within the T2DM group suggested that factors beyond genotype also influence ANRIL expression in T2DM patients. Collectively, these results suggested that the ANRIL gene could serve as a potential biomarker for T2DM, and further research was warranted to explore the specific mechanisms underlying its expression changes across different genotypes in the context of T2DM pathogenesis.

Fig. 2
figure 2

(A) LncRNA ANRIL level comparison. (B) Genotype level comparison of rs10757278 polymorphism in ANRIL. *** means P < 0.001

The serum ANRIL level was an independent influencing factor for the onset of T2DM

By employing logistic regression analysis to investigate the independent influencing factors of T2DM (Table 4), we found that age, TC, TG, HDL-C, LDL-C, SBP, and DBP were not significantly associated with the onset of T2DM (P > 0.05). However, BMI showed a significant correlation with the onset of T2DM, with an OR = 1.651, P = 0.037. Additionally, the rs10757278 variant exhibited a significant correlation with T2DM onset, with an OR = 1.833, and P = 0.029. Furthermore, ANRIL demonstrated an extremely significant correlation with T2DM onset, with an OR = 0.028, P < 0.001. In conclusion, BMI, the rs10757278 locus, and serum ANRIL levels were independent risk factors for the development of T2DM.

Table 4 Logistic regression analysis of factors related to the onset of T2DM

The ROC of serum ANRIL level in diagnosing T2DM

We conducted a detailed analysis and generated the ROC curve for ANRIL expression (Fig. 3), evaluating the model’s performance in predicting ANRIL expression-related conditions using this curve. The results demonstrated that the area under the curve (AUC) was 0.914, indicating a high level of predictive accuracy. Additionally, the sensitivity at a specific threshold was 88.72%, and the specificity was 80.36%. The high AUC value, along with favorable sensitivity and specificity, suggested that this model held significant potential for assessing ANRIL expression in T2DM. This provided a robust analytical tool for further investigation into the role of ANRIL in T2DM.

Fig. 3
figure 3

ROC of serum ANRIL level in the diagnosis of T2DM

Discussion

Diabetes impacts a substantial global population, transcending racial, gender, and age boundaries [25]. T2DM, the predominant form of diabetes, primarily results from impaired insulin function or secretion, or elevated hepatic glucose production [26]. Beyond the physiological mechanisms underlying diabetes, recent attention has focused on a class of molecules known as lncRNAs [27]. Emerging research indicates that lncRNA ANRIL is linked to several metabolic factors, including insulin resistance, glucose levels, inflammation, apoptosis, and aging [28]. Genetic polymorphisms play a crucial role in disease susceptibility and pathogenesis. The SNPs rs10757278 and rs10811656 exhibit significant associations with the expression levels of ANRIL (antisense non-coding RNA) and ANRIL in arterial thrombosis (AT) and peripheral blood mononuclear cells (PBMCs) [15]. However, the functional role of the ANRIL polymorphic locus rs10757278 in T2DM remains unclear. This study aimed to investigate the association between the ANRIL polymorphic locus rs10757278 and T2DM, elucidate the underlying mechanisms, and contribute to the development of personalized treatment strategies.

It has been demonstrated that certain SNPs linked to diseases exert their effects by modulating ANRIL itself [20]. To deepen our understanding of the molecular mechanisms underlying T2DM, it was essential to investigate the candidate genes involved in their onset and progression. This study aimed to explore the association between the rs10757278 polymorphic locus of ANRIL and T2DM. The study included 275 Chinese Han patients with T2DM and 280 healthy Chinese Han controls and analyzed the demographic and clinical characteristics of both groups. The results indicated that while there was no significant difference in age between the two groups, significant differences were observed in other parameters. Specifically, the BMI of the T2DM group was higher, and lipid profiles showed elevated levels of TC, TG, and LDL-C compared to the control group, while HDL-C was lower. Blood pressure measurements revealed that both SBP and DBP were higher in the T2DM group. Additionally, blood glucose indicators, including FPG and HbA1c, were significantly elevated in the T2DM group compared to the control group. Overall, significant differences were observed between T2DM patients and the control group in terms of BMI, lipid profiles, blood pressure, and blood glucose levels. Previous research has indicated that the G allele of the rs10757278 locus of ANRIL is associated with an increased risk of psoriasis in the studied population [29], and the rs10757278 A/G polymorphism is linked to overall cancer risk [30]. The genotypes AG and GG are associated with a smaller tumor size [31]. We analyzed the genotype and allele distribution of the polymorphic locus rs10757278 of ANRIL in patients with T2DM and the control group. Our findings revealed a significant difference in allele frequencies between the T2DM group and the control group. This suggested that this polymorphism may be linked to the onset and progression of T2DM. Specifically, the A allele was more prevalent in the control group and less frequent in the T2DM group, while the G allele showed the opposite trend. The G allele may be associated with an increased risk of T2DM. Moreover, the GG genotype was significantly associated with T2DM, with a higher proportion of GG genotype carriers observed in the T2DM group compared to the control group. Conversely, the proportion of AA genotype carriers was relatively lower in the T2DM group. Therefore, individuals carrying the GG genotype have a higher risk of developing T2DM, while the AG genotype may also be associated with T2DM, although this association has not yet reached statistical significance.

Genetic polymorphisms can influence the expression and function of genes [20, 22]. A detailed investigation into the demographic and clinical characteristics of patients with T2DM carrying various ANRIL rs10757278 genotypes was conducted. The findings revealed no significant differences in age, gender, BMI, TC, TG, HDL-C, SBP, and DBP among patients with different genotypes. However, significant differences were observed in LDL-C, FPG, and HbA1c. Specifically, the FPG and HbA1c levels were significantly affected by different genotypes. Carriers of the AA genotype exhibited relatively lower FPG and HbA1c levels compared to those with the GG genotype, indicating a distinct pattern of change. These results suggested that different genotypes influence blood glucose metabolism, providing valuable data for a deeper understanding of the genetic role in blood glucose regulation and serving as a foundation for future research and clinical applications. Additionally, the relationship between ANRIL gene expression and T2DM was explored. The study found that serum ANRIL levels in the T2DM group were significantly lower than in the control group, consistent with prior research showing reduced ANRIL expression in T2DM patients [32]. Furthermore, within both the control and T2DM groups, individuals with the AA genotype had higher serum ANRIL levels compared to those with the AG/GG genotypes. The differences in ANRIL levels across various genotypes were statistically significant, suggesting that this genetic polymorphism site might influence the pathogenesis of T2DM through the regulation of ANRIL levels. These findings imply that the ANRIL gene could play a crucial role in the development of T2DM. The considerable variability in relative ANRIL expression levels within the T2DM group indicates that, in addition to genotypes, other factors may also affect ANRIL gene expression in T2DM patients. ANRIL was hypothesized to impact cellular growth, differentiation, and survival by modulating intracellular signaling pathways. Specifically, it may disrupt intracellular signal transduction, altering the metabolic state and behavior of cells, and thus influencing the progression of T2DM. In summary, these results suggested that the ANRIL gene may serve as a potential biomarker for T2DM, and further investigation into the genotype-specific changes in its expression levels was warranted to elucidate its precise role in the disease’s pathogenesis.

Studies have demonstrated that the regulation of ANRIL expression influences susceptibility to T2DM [22]. Cellular senescence may confer a protective effect against T2DM, and lncRNA ANRIL is regarded as the most promising biomarker for cellular senescence in T2DM [32]. Logistic regression analysis revealed that factors such as age, gender, TC, TG, HDL-C, LDL-C, SBP, and DBP exhibited no significant association with the development of T2DM. In contrast, BMI, the rs10757278 locus, and serum ANRIL levels were identified as independent risk factors for T2DM. Further analysis of the ANRIL expression ROC curve indicated an AUC of 0.914, demonstrating high predictive accuracy. At a specific threshold, the sensitivity was 88.72% and the specificity was 80.36%. These findings suggest that serum ANRIL levels could serve as a potential diagnostic marker for T2DM. The high AUC value and favorable sensitivity and specificity indicated that this model held considerable promise in ANRIL expression for evaluating T2DM, offering an effective analytical tool for further investigations into the role of ANRIL in T2DM.

Previous studies have demonstrated that ANRIL is a key regulator of cellular senescence in T2DM [32]. ANRIL expression was found to be significantly upregulated in the peripheral venous blood of T2DM patients with myocardial infarction (MI) and in the myocardial tissue of T2DM-MI model mice [33]. However, no previous studies have established a definitive link between the rs10757278 polymorphism of the ANRIL gene and T2DM. Our study provided the first evidence confirming this association. In addition, this study has several limitations. The sample size was limited and the racial diversity is insufficient. Given that different racial and ethnic groups exhibit variations in genetic background, lifestyle, and dietary habits, these factors may influence the research outcomes. Future studies should develop a robust recruitment strategy to include a larger and more diverse population that meets the study criteria, thereby enhancing the generalizability of the findings. This approach will contribute to a more comprehensive understanding of the pathogenesis and influencing factors of T2DM.

In summary, the rs10757278 polymorphic site in lncRNA ANRIL was strongly associated with the genetic predisposition to T2DM. Studies have shown that individuals carrying the GG genotype were at a higher risk of developing T2DM. This association may be attributed to the GG genotype leading to a reduction in ANRIL levels. Further research has demonstrated that the serum levels of ANRIL in T2DM patients were significantly lower. This suggested that serum ANRIL levels could serve as a potential diagnostic marker for T2DM. The underlying mechanism may involve ANRIL’s role in regulating glucose metabolism. These findings contributed valuable insights into the pathogenesis of T2DM and offered promising avenues for early diagnosis and therapeutic interventions.

Data availability

Corresponding authors may provide data and materials.

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Acknowledgements

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Funding

This study was funded by The Science and Technology Project of the People’s Livelihood in Hebei Province (20377707D), Special Funding for Local Science and Technology Development Guided by the Central Government (206Z7701G) and the Key Projects of Hebei Administration of Traditional Chinese Medicine (Z2022015).

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Contributions

Study conception and design: X.Y. Li, A.G. Yang, X. Liu; data collection: R. Zhang, H.M. Zhou, S.J. Xu; analysis and interpretation of results: X.Y. Li, A.G. Yang, X. Liu, R. Zhang, H.M. Zhou, S.J. Xu; draft manuscript preparation: X.Y. Li, A.G. Yang, X. Liu, R. Zhang, H.M. Zhou, S.J. Xu. All authors reviewed the results and approved the final version of the manuscript.

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Correspondence to Huimin Zhou or Shunjiang Xu.

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Li, X., Yang, A., Liu, X. et al. Genetic association of long non-coding RNA ANRIL polymorphism with the risk of type 2 diabetes mellitus in the Chinese Han population. Diabetol Metab Syndr 17, 108 (2025). https://doi.org/10.1186/s13098-025-01670-3

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