Wang Y, Huang LH, Xu CX, Xiao J, Zhou L, Cao D, Liu XM, Qi Y. Connexin 32 and 43 promoter methylation in Helicobacter pylori-associated gastric tumorigenesis. World J Gastroenterol 2014; 20(33): 11770-11779 [PMID: 25206281 DOI: 10.3748/wjg.v20.i33.11770]
Corresponding Author of This Article
Can-Xia Xu, MD, Professor, Department of Gastroenterology, Third Xiangya Hospital of Central South University, 138 Tongzipo Street, Changsha 410013, Hunan Province, China. xucanxia2000@163.com
Research Domain of This Article
Gastroenterology & Hepatology
Article-Type of This Article
Original Article
Open-Access Policy of This Article
This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
World J Gastroenterol. Sep 7, 2014; 20(33): 11770-11779 Published online Sep 7, 2014. doi: 10.3748/wjg.v20.i33.11770
Table 1 Sex, age and disease duration of cases of each stage (mean ± SD)
Stage
n
M
F
Age, yr
Disease duration (yr)
NGM without H. pylori infection
25
14
11
54.12 ± 8.21 (45-60)
0.85 ± 0.64 (0.12-1.23)
NAG with H. pylori infection
24
13
11
56.44 ± 11.29 (47-65)
0.85 ± 0.35 (0.37-1.03)
CAG with H. pylori infection
25
11
14
55.90 ± 7.80 (45-66)
0.73 ± 0.24 (0.13-0.90)
IM with H. pylori infection
28
12
16
52.16 ± 8.59 (48-68)
0.90 ± 0.65 (0.47-1.04)
DYS with H. pylori infection
24
12
12
54.05 ± 7.36 (48-69)
0.73 ± 0.57 (0.56-1.15)
GC with H. pylori infection
30
17
13
55.43 ± 10.33 (46-73)
0.60 ± 0.46 (0.10-1.04)
Table 2Cx32 and Cx43 primer sequences, amplified fragment size and annealing temperature
Method
Gene
Primer sequence (5’→3’)
Amplified fragment size (bp)
Annealing temperature (°C)
Real-time RT-PCR
Cx32
F:
ATGAACTGGACAGGTTTGTAC
302
56
R:
ATGTGTTGCTGGTGCAGCCA
Cx43
F:
TGCAGCAGTCTGCCTTTCGTTG
219
56
R:
CCATCAGTTTGGGCAACCTTG
β-actin
F:
TGGACTTCGCAGCACAGCAGATGG
289
56
R:
ATCTCCTTCTGCATCCTGTCG
MSP
Cx32
M
F:
GGGGCGGGTGCGGCGAT
245
64
R:
CTCCGCGCCTACGTCCC
U
F:
GGGGTGGGTGTGGTGAT
245
64
R:
CTCCACACCTACATCCCAA
Cx43
M
F:
AAATTGTAATATTTGGGTTTCAGCGC
156
58
R:
AATAACGCCATCTCTACTCACCG
U
F:
TTTTAAAATTGTAATATTTGGGTTTCAGTGT
161
56
R:
AATAACACCATCTCTACTCACCACA
BSP
Cx32
F:
GGTTATTTTTTTGGTGGGGTTATG
313
58
R:
ACCCAAACAAATCCCCTATAATCTC
Cx43
F:
TGTTTTTTAAAATTGTAATATTTGGGTTTA
377
56
R:
AAAAACAAACTCATCTAACCTTCCTATTC
MassArray
Cx32
F:
CAGTTT CAGCAGTTTTTGGGTTTTTTGG
484
60
R:
TAACTCCCTATCCCCTAACTCCTTA
Cx43
F:
ATGTTTTTGCAGGTTGGATCAGGAAAT
447
60
R:
ACCAACAAATAAAAACAAAATTATTCC
Table 3Cx32 and Cx43 CpG loci detected by bisulfite polymerase chain reaction sequencing
Cx32
CpG
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
bp
28
34
37
45
49
83
86
120
132
155
227
231
247
253
255
○
○
○
○
●
●
●
●
●
●
●
●
●
●
●
Cx43
CpG
1
2
3
4
5
6
7
8
9
10
11
12
bp
32
34
37
44
71
127
134
143
159
190
198
280
○
○
○
○
●
●
●
●
●
●
●
●
Table 4Cx32 and Cx43 promoter methylation status at different gastric carcinogenesis stages with Helicobacter pylori infection by bisulfite polymerase chain reaction sequencing method
Table 5 Methylation levels (%) of Cx32 and Cx43 CpG islands at gastric different stages with Helicobacter pylori infection detected by MassArray method
Citation: Wang Y, Huang LH, Xu CX, Xiao J, Zhou L, Cao D, Liu XM, Qi Y. Connexin 32 and 43 promoter methylation in Helicobacter pylori-associated gastric tumorigenesis. World J Gastroenterol 2014; 20(33): 11770-11779