Copyright
©2005 Baishideng Publishing Group Inc.
World J Gastroenterol. Apr 7, 2005; 11(13): 1929-1936
Published online Apr 7, 2005. doi: 10.3748/wjg.v11.i13.1929
Published online Apr 7, 2005. doi: 10.3748/wjg.v11.i13.1929
Table 1 LC Protocol 1 for Genotyping the TGF-β1 Gene Polymorphisms.
Codon 10 (Leu10Pro) | ||||
Time (s) | Temp (°C) | Transition Rate (°C/s) | Cycles | |
600 | 95 | 20 | 1 | Denaturation |
10 | 95 | 20 | 45 | Cycling |
10 | 59 | 20 | ||
20 | 72 | 20 | ||
60 | 95 | 20 | 1 | Melting curve |
Analysis | ||||
30 | 45 | 20 | ||
0 | 78 | 0.2 | ||
5-d(CTAGGTTATTTCCGTGGG)-3’ | Sense primer | |||
5-d(CCTTGGCGTAGTAGTCG)-3’ | Reverse primer | |||
5-d(TCCCCCATGCCGCCCTCCGGGCT-Fluo)-3’ | Anchor primer | |||
5-LCRed640-(CGGCTGCTGCCGCTGCT-P)-3’ | Sensor primer |
Table 2 LC Protocol 2 for Genotyping the TGF-β1 Gene Polymorphisms.
Codon 25 (Arg25Pro) | ||||
Time (s) | Temp (°C) | Transition Rate (°C/s) | Cycles | |
600 | 95 | 20 | 1 | Denaturation |
10 | 95 | 20 | 45 | Cycling |
10 | 59 | 20 | ||
20 | 72 | 20 | ||
60 | 95 | 20 | 1 | Melting curve |
Analysis | ||||
30 | 40 | 20 | ||
0 | 80 | 0.2 | ||
5-d(CTAGGTTATTTCCGTGGG)-3’ | Sense primer | |||
5-d(CCTTGGCGTAGTAGTCG)-3’ | Reverse primer | |||
5-d(GCTACCGCTGCTGTGGCTACTGGTGCT-Fluo)-3 | Anchor primer | |||
5-LCRed640-(ACGCCTGGCCCGCCG-P)-3’ | Sensor primer |
Table 3 LC Protocol 3 for Genotyping the TGF-β1 Gene Polymorphisms.
Codon 263 (Thr263Ile) | ||||
Time (s) | Temp (°C) | Transition Rate (°C/s) | Cycles | |
600 | 95 | 20 | 1 | Denaturation |
10 | 95 | 20 | 45 | Cycling |
10 | 59 | 20 | ||
20 | 72 | 20 | ||
60 | 95 | 20 | 1 | Melting curve |
Analysis | ||||
30 | 39 | 20 | ||
0 | 74 | 0.2 | ||
5-d(AAGCAGGGTTCACTACCGGC)-3’ | Sense primer | |||
5-d(AGGCCTCCATCCAGGCTACA)-3’ | Reverse primer | |||
5-LCRed640-(GAGAGGGCCCAGCATCTGCAAAGCT-P)-3 | Anchor primer | |||
5-d(ATGGCCACCCCGCT-Fluo)-3’ | Sensor primer |
Table 4 TGF-β1 gene polymorphisms in subjects of Chinese and Caucasian origin.
Ethnic group | n | Subjects | Codon 25 | n | Codon 10 | n | Codon 263 | n |
Chinese | 194 | Controls | Arg/Arg | 104 | Leu/Leu | 29 | Thr/Thr | 104 |
Leu/Pro | 49 | |||||||
Pro/Pro | 26 | |||||||
Patients | Arg/Arg | 90 | Leu/Leu | 21 | Thr/Thr | 90 | ||
Leu/Pro | 38 | |||||||
Pro/Pro | 31 | |||||||
Caucasians | 260 | Controls | Arg/Arg | 42 | Leu/Leu | 19 | Thr/Thr | 19 |
Leu/Pro | 20 | Thr/Thr | 17 | |||||
Thr/Ile | 3 | |||||||
Pro/Pro | 3 | Thr/Thr | 2 | |||||
Thr/Ile | 1 | |||||||
Arg/Pro | 8 | Leu/Leu | 0 | |||||
Leu/Pro | 7 | Thr/Thr | 7 | |||||
Pro/Pro | 1 | Thr/Ile | 1 | |||||
Patients | Arg/Arg | 178 | Leu/Leu | 87 | Thr/Thr | 85 | ||
Thr/Ile | 2 | |||||||
Leu/Pro | 80 | Thr/Thr | 74 | |||||
Thr/Ile | 6 | |||||||
Pro/Pro | 11 | Thr/Thr | 7 | |||||
Thr/Ile | 4 | |||||||
Arg/Pro | 32 | Leu/Leu | 0 | |||||
Leu/Pro | 20 | Thr/Thr | 20 | |||||
Pro/Pro | 12 | Thr/Thr | 10 | |||||
Thr/Ile | 2 |
Table 5 TGF-β1 Allelic Frequencies.
Polymorphic site | Chinese | Caucasians | ||||
Total (n = 194) | Controls(n = 104) | Patients(n = 90) | Total(n = 260) | Controls(n = 50) | Patients (n = 210) | |
Codon 10 (T/C) | 0.50/0.50 | 0.51/0.49 | 0.44/0.56 | 0.65/0.35 | 0.65/0.35 | 0.65/0.35 |
Codon 25 (G/C) | 1/0 | 1/0 | 1/0 | 0.92/0.08 | 0.92/0.08 | 0.92/0.08 |
Codon 263 (C/T) | 1/0 | 1/0 | 1/0 | 0.96/0.04 | 0.95/0.05 | 0.97/0.03 |
Table 6 Genotype distribution of codon 10, 25, and 263 polymorphisms and grade of fibrosis in caucasian patients (n = 200) with chronic hepatitis C infection.
Codon | Genotype | Total | F0-2 | F3-4 |
10 | Leu/Leu | 82 | 70 | 12 |
Leu/Pro | 95 | 69 | 26 | |
Pro/Pro | 23 | 17 | 6 | |
25 | Arg/Arg | 169 | 137 | 32 |
Arg/Pro | 31 | 19 | 12 | |
Pro/Pro | 0 | 0 | 0 | |
263 | Thr/Thr | 186 | 145 | 41 |
Thr/Ile | 14 | 11 | 3 | |
Ile/Ile | 0 | 0 | 0 |
- Citation: Wang H, Mengsteab S, Tag CG, Gao CF, Hellerbrand C, Lammert F, Gressner AM, Weiskirchen R. Transforming growth factor-β1 gene polymorphisms are associated with progression of liver fibrosis in Caucasians with chronic hepatitis C infection. World J Gastroenterol 2005; 11(13): 1929-1936
- URL: https://www.wjgnet.com/1007-9327/full/v11/i13/1929.htm
- DOI: https://dx.doi.org/10.3748/wjg.v11.i13.1929