1. 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

2. Pin1 Wnt/β-catenin p53, p63, p73 ras/Neu-Jun NF-κB Cancer Upstream signals Signaling transduction Ser/Thr-Pro p Ser/Thr-Pro Ser/Thr-Pro Kinases Mapks GSKs CDKs Phosphalases PP2A

3. 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 11.61/10 5 during 1970-1972, increased to 32.67/10 5 during 1980-1982, to 41.54/10 5 during 1990-1992, and to 48.79/10 5 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.

4. 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.

5. 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

6. 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.

7. Genomic structure of PIN1 gene

8. 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

10. -667TT

13. Table II. PIN1 genotypes and allele frequencies and logistic regression analysis for associations with lung cancer risk

14. Patients (n = 1056)Controls (n = 1056) Crude OR ( 95% CI) Adjusted OR ( 95% CI) a P hom b GG n (%) GC+CC n (%) GG n (%) GC+CC n (%) GC+CC vs GG 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)0.852 > 60463(89.0)57(11.0)440(84.3)82(15.7)0.66(0.46-0.95)0.63(0.43-0.92) Sex Male672(90.1)74(9.9)634(85.0)112(15.0)0.62(0.46-0.85)0.62(0.45-0.86)0.837 Female276(89.0)34(11.0)261(84.2)49(15.8)0.66(0.41-1.05)0.71(0.43-1.16) Smoking Ever540(89.9)61(10.1)452(83.4)90(16.6)0.57(0.40-0.80)0.58(0.41-0.83)0.407 Never408(89.7)47(10.3)443(86.2)71(13.8)0.72(0.49-1.06)0.71(0.47-1.07) Drinking Ever208(90.8)21(9.2)187(82.4)40(17.6)0.47(0.27-0.83)0.50(0.28-0.92)0.366 Never740(89.5)87(10.5)708(85.4)121(14.6)0.69(0.51-0.92)0.69(0.51-0.93) Table III. Stratification analysis of the PIN1 -842G>C genotypes in lung cancer cases and controls

15. Patients (n = 1056)Controls (n = 1056) Crude OR ( 95% CI) Adjusted OR ( 95% CI) a P hom b GG n (%) GC+CC n (%) GG n (%) GC+CC n (%) GC+CC vs GG Family history of cancer Yes97(93.3)7(6.7)81(78.6)22(21.4)0.27(0.11-0.65)0.29(0.11-0.74)0.047 No851(89.4)101(10.6)814(85.4)139(14.6)0.70(0.53-0.91)0.70(0.53-0.93) BMI ≤23.9126(94.0)8(6.0)44(86.3)7(13.7)0.40(0.14-1.16)0.41(0.14-1.23)0.729 24.0-27.9730(89.0)90(11.0)593(84.5)109(15.5)0.67(0.50-0.91)0.66(0.49-0.89) ≥28.092(90.2)10(9.8)258(85.1)45(14.9)0.62(0.30-1.29)0.63(0.31-1.31) Table III. cont’ a ORs were adjusted by age, sex, smoking, drinking, BMI and family history of cancer b P value of the test for homogeneity between stratum-related ORs for Pin1-842.

16. All P<0.001 G-T G-C C-T C-C Luciferase assay: Pin1 -842 C variant decrease gene’s transcriptional activity

17. 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.

18. 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.

19. CONCLUSION The functional genetic variant -842G>C of Pin1 gene contributes to decreased the risk of lung cancer by diminishing the promoter activity

20. Limitations Hospital-based, retrospective study. Fewer SNPs were genotyped Restricted to a Chinese Han population

21. Dr. Lu’s lab staff, GZMC Dr. Wei’s lab staff, UT MDACCAcknowledgement

22. Thanks for your attention ! 谢谢！