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The polymorphisms and haplotypes
of Pin1 gene are associated with the
risk of lung cancer
吕嘉春,赵红军，杨磊，刘斌，纪卫东，宾晓农
广州医学院 化学致癌研究所、呼吸疾病国家重点实验室
承国家自然科学基金项目(30200235、30371196、30671813、30872178)资助
Upstream signals
Kinases
Ser/Thr-Pro
Mapks
GSKs
CDKs
Phosphalases
Ser/Thr-Pro
p Ser/Thr-Pro
PP2A
Signaling transduction
ras/Neu-Jun
p53, p63, p73
Pin1
NF-κB
Wnt/β-catenin
Cancer
BACKGROUND
 Lung cancer is one of the leading causes of death in the world, and
so does in Guangzhou.
 The death rate of lung cancer in Guangzhou city was
11.61/105 during 1970-1972, increased to 32.67/105 during 1980-1982,
to 41.54/105 during 1990-1992, and to 48.79/105 during 2000-2002.
 Pro-directed phosphorylation is an important signaling mechanism
controlling diverse cellular processes, including cell-cycle
progression, and cell proliferation and differentiation.
 The mechanisms controlling Pro-directed phosphorylation can result
in cell transformation and oncogenesis.
BACKGROUND
 Pin1 specifically regulates the conformation of Pro-directed
phosphorylation sites.
 Pin1 substrates include many essential cancer-related proteins, such
as von Hippel-Lindau tumor suppressor, myc, GSK -3 beta, cyclin D1,
Cdc25, cdc2/cyclin B, p73, and p53.
 Aberrant over expressions of Pin1 have been reported in many
cancers, including lung, breast and liver cancers.
 Inhibition of Pin1 in cancer cells can trigger apoptosis or suppress
the transformed phenotype. Moreover, the Pin1 knockout -/- female
mice showed impaired growth of mammary cells.
 The -842G>C polymorphism has been found to be associated with low risk of
head neck and breast cancer in American white population.
HYPOTHESIS
The genetic variations in the promoter
region of Pin1 gene and their possible
interaction with environmental factors
may play a role in the risk of lung cancer
STUDY DESIGN
 The studied SNPs were selected based on the dbSNP
database and re-sequencing data.
 A hospital-based case-control study 1056 newly
diagnosed patients with sporadic lung cancer were
recruited in Guangzhou.
 1056 cancer-free controls were recruited from healthy
subjects in the community health centers.
 The cases and controls frequency matched by age (±5
years) and sex
 All subjects were Chinese Han.
Genomic structure of PIN1 gene
GENOTYPING-PCR-RFLP
 Since the Pin1-842G>C (rs2233678) and -667T>C
(rs2233679) are close in distance, we used one pair of
primers
5’-CGG GCT CTG CAG ACT CTA TT -3’ (FP)
5’-AAA TTT GGC TCC TCC ATC CT -3’(RP)
Restriction enzyme: BanII for -842G>C
SacI for -667T>C
-667TT
Table I. Frequency distributions of selected variables in lung
cancer cases and controls
Variables
Age (years)
 60
> 60
Cases (n=1056)
n
%
Controls (n=1056)
n
%
pa
536
520
(50.8)
(49.2)
534
522
(50.6)
(49.4)
0.9306
746
310
(70.6)
(29.4)
746
310
(70.6)
(29.4)
1.0000
394
(37.3)
207
(19.6)
455
(43.1)
a
Two-sided 2 test;
366
176
514
(34.7)
(16.7)
(48.6)
Sex
male
female
Smoking
current
former
never
0.0283
Table I. cont’
Cases (n=1056)
Controls (n=1056)
Variables
Drinking
current
former
never
Family history
n
%
n
%
165
64
827
(15.6)
(6.1)
(78.3)
186
41
829
(17.6)
(3.9)
(78.5)
pa
0.0429
0.9417
yes
no
BMI
<18
18-25
>25
Sub-ethnic group
TeoChewese(潮汕)
Hakka (客家)
Cantonese（广府）
Other Hans(他省汉)
104
952
(9.9)
(90.1)
103
953
(9.8)
(90.2)
134
820
102
(12.7)
(77.6)
(9.7)
51
702
303
(4.8)
(66.5)
(28.7)
<0.0001
90
198
683
85
(8.5)
(18.8)
(64.7)
(8.0)
81
195
700
80
(7.7)
(18.5)
(66.3)
(7.5)
0.8358
Table II. PIN1 genotypes and allele frequencies and logistic regression
analysis for associations with lung cancer risk
Genotypes
-842G>C
GG
GC
CC
P trend
GC+CC
C allele
-667C>T
CC
CT
TT
P trend
CT+TT
T allele
Haplotypes
G-T
G-C
C-T
C-C
Cases
n (%)
Controls
n (%)
948(89.8)
103(9.7)
5(0.5)
895(84.7)
154(14.6)
7(0.7)
108(10.2)
0.053
161(15.3)
0.079
367 (34.7)
512 (48.5)
177 (16.8)
352 (33.3)
522 (49.4)
182 (17.3)
689 (65.3)
0.410
704 (66.7)
0.420
863(40.9)
1136(53.8)
39(1.8)
74(3.5)
832(39.4)
1112(52.6)
54(2.6)
114(5.4)
Pa
Crude
OR (95% CI)
Adjusted
OR (95% CI) b
0.0025
1.00 (ref.)
0.63(0.48-0.83)
0.68(0.21-2.14)
0.0008
0.64(0.49-0.83)
1.00 (ref.)
0.64(0.48-0.84)
0.77(0.23-2.52)
0.0018
0.64(0.49-0.84)
1.00 (ref.)
0.94(0.78-1.14)
0.93(0.72-1.20)
0.6179
0.94(0.78-1.12)
1.00 (ref.)
0.89(0.73-1.08)
0.83(0.64-1.08)
0.6984
0.87(0.73-1.06)
1.00 (ref)
0.97(0.75-1.26)
0.50(0.17-1.49)
0.40(0.20-0.78)
1.00 (ref)
1.11(0.85-1.46)
0.64(0.21-1.95)
0.41(0.21-0.82)
0.0007
0.5132
0.5322
0.007
Two-sided 2 test for either genotype distribution or allele frequency.
b
Adjusted for age, sex, smoking status, and alcohol, family cancer history.
a
Table III. Stratification analysis of the PIN1 -842G>C
genotypes in lung cancer cases and controls
Patients (n = 1056)
GG
n (%)
GC+CC
n (%)
Controls (n = 1056)
Crude OR
( 95% CI)
Adjusted OR
( 95% CI)a
GC+CC vs GG
GG
n (%)
GC+CC
n (%)
GC+CC vs GG
Phomb
Age (years)
 60
485(90.5)
51(9.5)
455(85.2)
79(14.8)
0.61(0.42-0.88)
0.65(0.44-0.95)
> 60
463(89.0)
57(11.0)
440(84.3)
82(15.7)
0.66(0.46-0.95)
0.63(0.43-0.92)
Male
672(90.1)
74(9.9)
634(85.0)
112(15.0)
0.62(0.46-0.85)
0.62(0.45-0.86)
Female
276(89.0)
34(11.0)
261(84.2)
49(15.8)
0.66(0.41-1.05)
0.71(0.43-1.16)
Ever
540(89.9)
61(10.1)
452(83.4)
90(16.6)
0.57(0.40-0.80)
0.58(0.41-0.83)
Never
408(89.7)
47(10.3)
443(86.2)
71(13.8)
0.72(0.49-1.06)
0.71(0.47-1.07)
Ever
208(90.8)
21(9.2)
187(82.4)
40(17.6)
0.47(0.27-0.83)
0.50(0.28-0.92)
Never
740(89.5)
87(10.5)
708(85.4)
121(14.6)
0.69(0.51-0.92)
0.69(0.51-0.93)
0.852
Sex
0.837
Smoking
0.407
Drinking
0.366
Table III. cont’
Patients (n = 1056)
GG
n (%)
GC+CC
n (%)
Controls (n = 1056)
GG
n (%)
GC+CC
n (%)
Crude OR
( 95% CI)
GC+CC vs
GG
Adjusted OR
( 95% CI)a
Phomb
GC+CC vs GG
Family
history of
cancer
Yes
97(93.3)
7(6.7)
81(78.6)
22(21.4)
0.27(0.11-0.65)
0.29(0.11-0.74)
No
851(89.4)
101(10.6)
814(85.4)
139(14.6)
0.70(0.53-0.91)
0.70(0.53-0.93)
≤23.9
126(94.0)
8(6.0)
44(86.3)
7(13.7)
0.40(0.14-1.16)
0.41(0.14-1.23)
24.0-27.9
730(89.0)
90(11.0)
593(84.5)
109(15.5)
0.67(0.50-0.91)
0.66(0.49-0.89)
≥28.0
92(90.2)
10(9.8)
258(85.1)
45(14.9)
0.62(0.30-1.29)
0.63(0.31-1.31)
0.047
BMI
a
b
ORs were adjusted by age, sex, smoking, drinking, BMI and family history of cancer
P value of the test for homogeneity between stratum-related ORs for Pin1-842 .
0.729
Luciferase assay: Pin1 -842 C variant decrease gene’s transcriptional activity
G-T
G-C
C-T
Luciferase activity
(Relative level)
C-C
G-T
G-C
C-T
C-C
350
300
250
200
150
100
50
0
A549
L78
All P<0.001
16HBE
Population stratification
• Multilevel Logistic regression null model:
the sub-ethnic group to the risk of lung cancer
OR=0.971，95%CI=0.862–1.094；P=0.6277
• Multilevel Logistic regression 2 level model:
the sub-ethnic group as level 2, age, sex, smoking,
drinking and Pin1-842G>C genotypes as level 1,
Pin1-842G>C: OR=0.639, 95%C.I.=0.488–0.836；
P=0.0011.
After controlling the confounding effect of sub-ethnic
groups, the main effect of Pin1-842G>C is still
significant.
False positive report probability (FPRP) for association between
Pin1 -842 G>C polymorphism with lung cancer risk
OR=0.64 (0.49-0.84), Power=0.927
null OR=0.67 (or 1/1.5)
Prior probability =0.01
FPRP=0.06 → noteworthy at 0.2 level
The finding of this study is unlikely by chance.
CONCLUSION
The functional genetic variant -842G>C
of Pin1 gene contributes to decreased
the risk of lung cancer by diminishing
the promoter activity
Limitations
• Hospital-based, retrospective study.
• Fewer SNPs were genotyped
• Restricted to a Chinese Han population
Acknowledgement
Dr. Lu’s lab staff, GZMC
Dr. Wei’s lab staff, UT MDACC
Thanks for your attention !
谢谢！