Manuscript accepted on :21-09-2022
Published online on: 08-11-2022
Plagiarism Check: Yes
Reviewed by: Dr. Ian Martins
Views: (Visited 265 times, 1 visits today)
PDF Downloads:
215Hussein Najm Abed 1*
, Ahmed Jalal Mohammed Albayati 2 , Muthana Salih Mashkor3, and Abdullah Shakir4
1Medical Laboratory Technology Department, Altoosi University College, Najaf, Iraq.
2College of pharmacy , Kufa University, Najaf, Iraq.
3College of Science, Kufa University, Najaf, Iraq.
4The Islamic University, Najaf, Iraq
Corresponding Author E-mail: hussein.najm@altoosi.edu.iq
DOI : https://dx.doi.org/10.13005/bpj/2578
Abstract
The widespread prevalence of diabetes has led to a serious health risk for humans. Hyperglycemia is a hallmark of diabetes. Assessing the relation of TCF7L2 gene polymorphism (rs7903146) on the occurrence of T2DM in the Alnajaf population, and evaluating the impact of this investigated SNP on phenotypic parameters like BMI, fasting blood sugar(F.B.S), lipid profile , Mg and Zn. A case-control study of 800 individuals (400 T2DM and 400 apparently healthy control individuals) was done. The laboratory work was carried out in the laboratory of Department of Clinical Laboratory Sciences in the Faculty of Pharmacy and in the laboratory of Department of Chemistry in the Faculty of Sciences / the University of Kufa. DNA from whole blood was extracted and genotyping of TCF7L2 gene (rs7903146) polymorphism polymorphisms were carried out by RFLP –PCR. The results of this study showed that there is a significant association between the transcription factor 7-like 2 (TCF7L2) gene polymorphisms rs7903146 gene polymorphism with T2DM in the Alnajaf population. TCF7L2 gene polymorphism (rs7903146) did not show any major changes in relation to BMI, FSG, cholesterol, HDL-C, VLDL-C, or T.G, but there were significant changes in LDL, magnesium, and zinc.
Keywords
Al-Najaf; Diabetes Mellitus; type 2; Polymorphism; Transcription Factor;TCF7L2
Download this article as:
| Copy the following to cite this article: Abed H. N, Albayati A. J. M, Mashkor M. S, Shakir A. Association of Transcription Factor 7 Like 2 (TCF7L2) ( rs7903146) Gene Polymorphisms with Some Trace Elements in the Type 2 Diabetes Mellitus Patients in Al-Najaf Governorate. Biomed Pharmacol J 2022;15(4). |
| Copy the following to cite this URL: Abed H. N, Albayati A. J. M, Mashkor M. S, Shakir A. Association of Transcription Factor 7 Like 2 (TCF7L2) ( rs7903146) Gene Polymorphisms with Some Trace Elements in the Type 2 Diabetes Mellitus Patients in Al-Najaf Governorate. Biomed Pharmacol J 2022;15(4). Available from: https://bit.ly/3FV8Yda |
Introduction
Chronic hyperglycemia due to changes in insulin secretion, insulin action, or both characterizes the metabolic diseases known collectively as diabetes mellitus. Insulin’s role as an anabolic hormone contributes to metabolic abnormalities in carbs, fats, and proteins 1. These metabolic disorders are caused by insulin resistance and/or low levels of insulin at the receptor, signal transduction system, or effector enzyme or gene levels in target tissues, notably skeletal muscles and adipose tissue, and to a lesser extent liver 2. Approximately 415 million adults worldwide have diabetes, and nearly half of them (46.5%) have never been diagnosed. Every six seconds, someone dies from diabetes (5.0 million deaths per year) 3. About 642 million people in the United States will have diabetes by the year 2040. The majority of the $673 billion spent annually on healthcare around the world goes toward diabetes care (12 percent) 4.
The TCF7L2 gene has received a lot of attention because of the strong correlation between the two (T2DM) 5. Originally connected with development biology, the Wnt signaling pathway is a complex network of interacting proteins that regulates cellular intercommunications at multiple levels and has a wide variety of consequences. The TCF7L2 gene is a key component of this route 6. TCF7L2 is just one example of a Wnt network-associated protein that has been related to a wide range of diseases and cancer models, demonstrating the importance of this developmental pathway in the pathogenesis of human disorders 7.
The TCF7L2 gene’s frequent polymorphism, rs7903146(C/T), is linked to an increased risk of type 2 diabetes if you have the riskier T allele 8. The TCF7L2 gene product, a high mobility group box-containing transcription factor previously related in blood glucose management, is assumed to be involved in Wnt signaling’s regulation of proglucagon gene expression in enteroendocrine cells 9. There is a positive correlation between TCF7L2 expression and insulin gene expression in human islets 10.
Consequently, we set out to investigate the associations between polymorphisms in the Transcription factor 7 like 2 gene (TCF7L2) and type 2 diabetes, as well as the effects of this association on biochemical and clinical outcomes.
Materials and Methods
Collection of Samples
Four hundred people with type 2 diabetes participated in the study (200 male and 200 female). The ages of our patients ranged from 40 to 70, with a mean sd of 52.93 7.00. Four hundred participants who were otherwise healthy were assigned to the “healthy” group (200 male and 200 female). AL-Sadr Medical City in al Najaf Province is where they were selected from
PCR Assay
The Intron kit makes blood DNA extraction and purification much easier. High-purity strands of DNA have been successfully employed in agarose gel electrophoresis, restriction enzyme digestion, and polymerase chain reaction (PCR). There are four fundamental stages to this procedure. Methods for Isolating Genomic Information from Frozen Blood Samples Consistent with the instructions provided in the Intron Kit ( iNtRoN, Biotech. Inc., Korea). As part of the gel documentation procedure (Cleaver, United Kingdom), 1% agarose (iNtRoN, Biotech, Inc., Korea) was used to examine and disperse the migration of PCR bands, and the gel was back-dyed with ethidium bromide at a concentration of 0.5 g/ml. These studies employ RFLP-PCR, and the primer sequence and restriction enzyme employed in the experiment are listed in the table below.
Table 1
| Gene | Primer | Sequence | Tm(ºC) |
| TCF7L2
rs7903146 |
Forward | 5′-TTA GAG AGC TAA GCA CTT TTT AGG TA-3′ | 61.6 |
| Reverse | 5′-AGA GAT GAA ATG TAG CAG TGA AGT G-3′ | 62.5 |
Statistical Analysis
The SPSS V.25 program was used in statistical analysis of the data. To analyze the disparities between these factors, the chi-square test was applied. Continuous parameters’ means were compared among genotypes using the ANOVA test.
Results and Discussion
Genetic Polymorphism of TCF7L2 rs7903146
The PCR products of TCF7L2 gene SNP (rs79103146) are 175bp
 |
Figure 1: PCR product of TCF7L2 gene polymorphism (rs79103146) analyzed by agarose gel electrophoresis. |
RFLP analysis for TCF7L2 gene
Restriction enzymes were used for digestion of TCF7L2 amplicon the resulted products were subjected to electrophoresis as in the following:
 |
Figure 2: Digested products in agarose electrophoresis for TCF7L2 gene (rs7903146) polymorphism. |
Table 2: Results for digested (rs7903146) polymorphism of TCF7L2 gene.
| Genotyping | Bands number | Size | |
| Wild | CC | 2 | 20 and 80 |
| Hetero | CT | 3 | 20,80, and 175 |
| Homo | TT | 1 | 175 |
Table 3: Genotyping and frequency of alleles of TCF7L2(rs7903146) polymorphism in studied group
| C/T | Control (n= 400) | T2DM (n= 400) | Unadjusted OR (95% CI)
P-Value |
adjusted OR (95% CI)
P-Value |
| Codominant | ||||
| CC | 260 | 185 | REF | REE |
| CT | 115 | 172 | 2.10
1.55-2.84 P < 0.0001 |
3.13
2.05-4.76 P < 0.0001 |
| TT | 25 | 43 | 2.42
1.43 – 4.10 0.001 |
2.57
1.51-4.38 P < 0.0001 |
| Dominate | ||||
| CT+ TT | 140 | 215 | 2.16
1.62- 2.87 P < 0.0001 |
2.34
1.75-3.12 P < 0.0001 |
| Recessive | ||||
| CC + CT(REF) | 375 | 357 | ——– | ——– |
| TT | 25 | 43 | 1.81
1.08 – 3.02 0.024 |
1.87
1.12-3.15 0.018 |
| Additive | ||||
| 2(TT)+CT | 165 | 258 | 2.31
1.76 to 3.04 P < 0.0001 |
2.83
1.95-4.12 P < 0.0001 |
| Frequency of T allele % |
20.6 % | 31% | 2.10
1.50 – 2.94 P < 0.0001 |
2.26
1.61-3.18 P < 0.0001 |
Table 4 : The relation between TCF7L2 (rs7903146 C>T) gene polymorphism genotypes and the investigated parameters in patients’ group under the codominant model.
| Clinical Characteristic | Genotype M ± SD | P value | |||
| CC CT | CT TT | ||||
| CC(n=185) | CT (n=172) | TT(n=43) | CC TT | ||
| BMI | 29.34 ±5.08 | 28.41 ± 5.15 | 28.10±5.09 | 0.089 | 0.154
|
| 0.724 | |||||
| FPG (mg/dl) | 181.58 ± 63.59 | 184.99 ± 78.28 | 194.47 ± 93.89 | 0.663 | 0.452 |
| 0.303 | |||||
| Cholesterol (mg/dl) | 269.17 ± 26.90 | 269.05 ± 26.94 | 270.05 ± 27.25 | 0.965 | 0.828 |
| 0.848 | |||||
| HDL-C (mg/dl) | 37.71± 6.45 | 36.88 ± 7.21 | 36.74± 7.32 | 0.251 | 0.905 |
| 0.402 | |||||
| Triglycerides(mg/dl) | 242.84 ± 47.70 | 247.98 ± 48.70 | 247.01 ± 47.97 | 0.314 | 0.906 |
| 0.609 | |||||
| VLDL (mg/dl) | 48.57± 9.54 | 49.6 ± 9.74 | 49.4 ± 9.59 | 0.314 | 0.906 |
| 0.609 | |||||
| LDL (mg/dl) | 162.39 ± 34.76 | 319.0 ± 173.18 | 311.10 ± 171.51 | < 0.0001 | 0.719 |
| < 0.0001 | |||||
| Mg (mg/dl) | 1.4588 ± 0.33551 | 1.2107 ± 0.36692 | 1.1999 ± 0.35733 | < 0.0001 | 0.857 |
| < 0.0001 | |||||
| Zn (mg/dl) | 81.05 ± 25.77 | 58.16± 26.70 | 50.60 ± 28.16 | < 0.0001 | 0.094 |
| < 0.0001 | |||||
| < 0.0001 | |||||
| 0.094 | |||||
Table 5: The relation between TCF7L2 (rs7903146 C>T) gene polymorphism genotypes and the investigated parameters in patients’ group under the dominant model.
| Clinical Characteristic | Genotype M ± SD | p-value | |
| CC(n=185) | CT + TT(n=215) | ||
| BMI | 28.18 ± 5.0 | 29.24 ± 5.0 | 0.037 |
| FSG (mg/dl) | 181.56 ± 63.59 | 183.93 ± 77.22 | 0.743 |
| Cholesterol(mg/dl) | 248.67 ± 33.32 | 403.83 ± 172.16 | < 0.0001 |
| HDL (mg/dl) | 37.71 ± 6.46 | 36.85 ± 7.22 | 0.0210 |
| Triglycerides(mg/dl) | 242.84 ± 47.7 | 247.78± 48.4 | 0.306 |
| VLDL (mg/dl) | 48.57 ± 9.5 | 49.5 ± 9.7 | 0.306 |
| LDL (mg/dl) | 162.4 ± 34.76 | 317.42 ± 172.47 | < 0.0001 |
| Mg (mg/dl) | 1.45 ± 0.33 | 1.2 ± 0.36 | < 0.0001 |
| Zn (mg/dl) | 53.94±24.43 | 79.83± 27.47 | < 0.0001 |
There were no significant differences in the distributions of TCF7L2 genotypes when comparing body mass index (B.M.I), fasting glucose, cholesterol, high-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, or total cholesterol to triglyceride levels or triglyceride levels with 12 but there were significant changes in LDL, magnesium, and zinc.
Variations in body mass index (BMI), cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), magnesium, and zinc all show substantial differences when compared to the dominant genotype distribution.
Low insulin levels, reduced waist circumference, and better lipid profiles were associated with the TCF7L2 rs7903146 polymorphism in the general senior population of Tuscany, Italy. Patients with diabetes who carry the minor allele are at reduced risk for developing metabolic syndrome 13.
Conclusions
TCF7L2 gene (rs7903146) polymorphism, was correlated with the tendency to T2DM in Alnajaf populations. TCF7L2 gene polymorphism (rs7903146) did not show any major changes in relation to BMI, FSG, cholesterol, HDL, VLDL, or T.G, but there were significant changes in LDL, magnesium, and zinc.
Recommendations
1-Study other SNPs of TCF7L2 gene to assess their association with T2DM.
2-Sequencing of the examined gene in order to find new SNPs associated with T2DM.
3-Haplotype study of the gene investigated to determine their association with type 2 diabetes.
5-Analysis of more SNPS of TCF7L2 gene to determine which one is more common in our population.
6-Evaluation of gene expression to determine the effect of SNPS on the various phenotypic properties.
7-DNA sequencing may give more effective results in regard to the correlation of SNPS on the various phenotypic parameters in type 2 diabetic patients.
Conflict of Interest
References
- Priyanka DN, Prashanth KVH, Tharanathan RN. A review on potential anti-diabetic mechanisms of chitosan and its derivatives. Carbohydr Polym Technol Appl. Published online 2022:100188.
CrossRef - Balaji R, Duraisamy R, Kumar MP. Complications of diabetes mellitus: A review. Drug Invent Today. 2019;12(1).
- Agarwal MM. Gestational diabetes in the Arab gulf countries: Sitting on a land-mine. Int J Environ Res Public Health. 2020;17(24):9270.
CrossRef - Ng LC, Gupta M. Transdermal drug delivery systems in diabetes management: A review. Asian J Pharm Sci. 2020;15(1):13-25.
CrossRef - Huang Z, Liao Y, Huang R, Chen J, Sun H. Possible role of TCF7L2 in the pathogenesis of type 2 diabetes mellitus. Biotechnol Biotechnol Equip. 2018;32(4):830-834.
CrossRef - del Bosque-Plata L, Martínez-Martínez E, Espinoza-Camacho MÁ, Gragnoli C. The role of TCF7L2 in type 2 diabetes. Diabetes. 2021;70(6):1220-1228.
CrossRef - Sharma A, Mir R, Galande S. Epigenetic regulation of the Wnt/β-catenin signaling pathway in cancer. Front Genet. 2021;12.
CrossRef - Ding W, Xu L, Zhang L, et al. Meta-analysis of association between TCF7L2 polymorphism rs7903146 and type 2 diabetes mellitus. BMC Med Genet. 2018;19(1):1-12.
CrossRef - Xu L. Transcription Factor 7-like 2 (TCF7L2) Gene Polymorphisms in Relation to the Risk of Type 2 Diabetes in three ethnicities. Published online 2018.
- Grant SFA. The TCF7L2 locus: a genetic window into the pathogenesis of type 1 and type 2 diabetes. Diabetes Care. 2019;42(9):1624-1629.
CrossRef - Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, Van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990;28(3):495-503.
CrossRef - Assmann TS, Duarte GCK, Rheinheimer J, Cruz LA, Canani LH, Crispim D. The TCF7L2 rs7903146 (C/T) polymorphism is associated with risk to type 2 diabetes mellitus in Southern-Brazil. Arq Bras Endocrinol Metabol. 2014;58(9):918-925.
CrossRef - Melzer D, Murray A, Hurst AJ, et al. Effects of the diabetes linked TCF7L2 polymorphism in a representative older population. BMC Med. 2006;4(1):1-8.
CrossRef
