Serum MicroRNAs as Predictors for HCV Progression and Response to Treatment in Pakistani Patients
Abstract
:1. Introduction
2. Materials and Methods
2.1. Data and Sample Collection
2.2. Parameter Analysis
- All of the chemical parameters related to the liver were done using a kit method based on spectrophotometry.
- A commercial ELISA kit determined α fetoproteins.
- RT PCR determined HCV RNA (viral load).
2.3. Expression Analysis
2.4. cDNA Synthesis by Reverse Transcriptase
2.5. PCR for Expression Analysis of Selected miRNA Genes (miRNA-182, miRNA-122, mi-RNA21, miRNA-199, miRNA-150, and miRNA-222)
2.6. Statistical Analysis
3. Results
3.1. Demographics Variables
3.2. Association Analysis
3.3. Analysis of Variance
3.4. Association Analysis
3.5. Expression Analysis of miRNAs in Diseased and Control Groups Using Real-Time PCR
3.6. Expression Analysis of miRNA in Chronic Patient and Treated Groups Using Real-Time PCR
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer Name | Primer Sequence |
---|---|
miRNA-182-F | 5′-TGCGGTTTGGCAATGGTAGAAC-3’ |
miRNA-182-R | 5’-CCAGTGCAGGGTCCGAGGT-3’ |
miRNA-122–F | 5′-ACACTCCAGCTGGGTGGAGTGTGACAATCC-3′ |
miRNA-122R | 5′-TGGTGTCGTGGAGTCG-3′ |
miRNA-21-F | 5′-GCCCGCTAGCTTATCAGACTGATG-3′ |
miRNA-21-R | 5′-CAGTGCAGGGTCC GAGGT-3 |
miRNA-199–F | 5′-GCGGCGGACAGTAGTCTGCAC-3′ |
miRNA-199–R | 5′-ATCCAGTGCAGGGTCCGAGG-3′ |
miRNA-150-F | 5′TCTCCCAACCCTTGTACCAGTG3′ |
miRNA-150-R | 5′CAGTGCGTCGTGGAGT3′ |
miRNA-222-F | 5′-GTTCGTGGGAGCTACATTGTCTGC-3′ |
miRNA-222-R | 5′-GTGTCGTGGAGTCGGCAATTC-3′ |
U6 snRNA-F | CTCGCTTCGGCAGCACATATAC |
U6 snRNA-R | ACGCTTCACGAATTTGCGTGTC |
Variables | Chronic (n = 40) | Cirrhosis (n = 40) | HCC (n = 40) | Treated (n = 40) | Control (n = 40) | p Value | |
---|---|---|---|---|---|---|---|
Gender | Male | 20 (50%) | 20 (50%) | 30 (75%) | 18 (45%) | 20 (50%) | 0.057 |
Female | 20 (50%) | 20 (50%) | 10 (25%) | 22 (55%) | 20 (50%) | ||
Age | 21–40 | 22 (55%) | 8 (20%) | 1 (2.5%) | 10 (25%) | 30 (75%) | 0.001 |
41–60 | 15 (37.5%) | 29 (72.5%) | 29 (72.5%) | 26 (65%) | 6 (15%) | ||
61–80 | 3 (7.5%) | 3 (7.5%) | 10 (25%) | 4 (10%) | 4 (10%) |
Male (n = 108) | Female (n = 92) | p Value | |
---|---|---|---|
Variables | M ± SD | M ± SD | |
Age | 48.96 ± 11.96 | 44.48 ± 10.65 | 0.006 |
ALT | 35.18 ± 17.50 | 32.48 ± 11.08 | 0.20 |
AST | 34.43 ± 15.69 | 31.34 ± 10.74 | 0.11 |
ALP | 175.05 ± 105.46 | 157.52 ± 57.62 | 0.16 |
Viral Load | 3,952,923.66 ± 34,763,237.74 | 1,081,468.79 ± 4,053,847.94 | 0.43 |
Bilirubin | 1.01 ± 0.60 | 0.88 ± 0.55 | 0.11 |
Variables | Chronic (n = 40) | Cirrhosis (n = 40) | HCC (n = 40) | Control (n = 40) | p Values |
---|---|---|---|---|---|
Age | 41.35 ± 11.55 | 49.08 ± 9.11 | 56.85 ± 6.62 | 38.50 ± 10.86 | 0.000 |
ALT | 35.38 ± 15.70 | 35.13 ± 12.38 | 43.08 ± 20.42 | 29.05 ± 3.81 | 0.000 |
AST | 33.13 ± 12.50 | 32.90 ± 12.74 | 41.65 ± 21.83 | 27.88 ± 3.04 | 0.000 |
ALP | 179.53 ± 45.38 | 163.70 ± 68.85 | 210.85 ± 137.64 | 91.65 ± 19.83 | 0.000 |
Viral Load | 11,191,265.55 ± 57,129,256.41 | 735,073.68 ± 1,357,141.14 | 1,233,932.88 ± 1,946,918 | 0.00 ± 0.00 | 0.25 |
Bilirubin | 0.77 ± 0.26 | 0.94 ± 0.48 | 1.65 ± 0.75 | 0.64 ± 0.20 | 0.000 |
AFP | 808.15 ± 1381.09 | NA |
Chronic (n = 40) | Treated (n = 40) | p Value | |
---|---|---|---|
Variables | M ± SD | M ± SD | |
Age | 41.35 ± 11.55 | 48.73 ± 9.52 | 0.003 ** |
ALT | 35.38 ± 15.70 | 31.63 ± 4.67 | 0.15 |
AST | 33.13 ± 12.50 | 29.50 ± 5.98 | 0.10 |
ALP | 184.48 ± 35.17 | 189.20 ± 63.56 | 0.68 |
Viral Load | 11,191,265.55 ± 57,129,256.41 | 0.00 ± 0.00 | 0.22 |
Bilirubin | 0.77 ± 0.26 | 0.77 ± 0.39 | 0.95 |
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Manzoor, S.; Malik, I.R.; Jahan, S.; Sarwar, M.B.; Bashir, A.; Shams, S.; Hussain, A. Serum MicroRNAs as Predictors for HCV Progression and Response to Treatment in Pakistani Patients. Genes 2023, 14, 441. https://doi.org/10.3390/genes14020441
Manzoor S, Malik IR, Jahan S, Sarwar MB, Bashir A, Shams S, Hussain A. Serum MicroRNAs as Predictors for HCV Progression and Response to Treatment in Pakistani Patients. Genes. 2023; 14(2):441. https://doi.org/10.3390/genes14020441
Chicago/Turabian StyleManzoor, Sadia, Imran Riaz Malik, Shah Jahan, Muhammad Bilal Sarwar, Asma Bashir, Sulaiman Shams, and Abrar Hussain. 2023. "Serum MicroRNAs as Predictors for HCV Progression and Response to Treatment in Pakistani Patients" Genes 14, no. 2: 441. https://doi.org/10.3390/genes14020441
APA StyleManzoor, S., Malik, I. R., Jahan, S., Sarwar, M. B., Bashir, A., Shams, S., & Hussain, A. (2023). Serum MicroRNAs as Predictors for HCV Progression and Response to Treatment in Pakistani Patients. Genes, 14(2), 441. https://doi.org/10.3390/genes14020441