doi: 10.1111/jop.12211
J Oral Pathol Med © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd wileyonlinelibrary.com/journal/jop
Genetic variants in AKT1 gene were associated with risk and survival of OSCC in Chinese Han Population Yun Wang1,†, Lin Lin1,†, Hao Xu1,2, Taiwen Li1, Yu Zhou1, Hongxia Dan1, Lu Jiang1, Ga Liao1, Min Zhou1, Longjiang Li1, Xin Zeng1, Jing Li1, Qianming Chen1 1
State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China; 2West China School of Public Health, Sichuan University, Chengdu, China
BACKGROUND: AKT1 is an important downstream effector of PTEN/PI3K/AKT signal transduction pathway. Aberrant expression and genetic variant of AKT1 gene are suggested to be involved in several types of human cancers, including OSCC. The aim of this study was to investigate the possible association between AKT1 gene polymorphisms and OSCC in Chinese Han Population. METHODS: A total of 182 OSCC patients and 207 cancer-free controls were enrolled for this hospital-based study. Five single-nucleotide polymorphisms (SNPs) on AKT1 (rs1130214, rs1130233, rs2494732, rs3730358, rs3803300) were investigated and genotyped by Sequenom Mass ARRAY & iPLEX-MALDI-TOF technology. Chi-square test, SHEsis software, and Kaplan–Meier method were used to evaluate the relationship between selected SNPs and OSCC susceptibility and progression. RESULTS: Significant difference of genotype distribution was observed between cases and control group at SNP sites rs1130214 (P = 0.006) and rs3803300 (P = 0.033, P = 0.003 for heterozygote and homozygous mutant, respectively). In the haplotype analysis, haplotype H4 which contained mutant-type allele of rs1130214 and rs3803300 was also related to OSCC risk (OR = 1.974, 95% CI = 1.048–3.718). Moreover, CT genotype of rs3730358 was associated with higher risk of OSCC progression (HR = 2.466, 95% CI = 1.017–5.981). CONCLUSION: Our results indicated that rs1130214 and rs3803300 were related to OSCC susceptibility in Chinese Han Population. In addition, rs3730358 might be associated with progression-free survival time of OSCC patients, suggesting that this SNP could be a potential prognosis marker for OSCC. J Oral Pathol Med (2014)
Correspondence: Jing Li and Xin Zeng, State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, No.14, Section 3, Renmin Nan Road, Chengdu 610041, China. Tel: +8615928152276, Fax: +86-02885501484, E-mails: [email protected] and [email protected] † These authors contributed equally to this paper. Accepted for publication April 28, 2014
Keywords: AKT1; oral squamous cell carcinoma; single-nucleotide polymorphism; survival; susceptibility
Introduction Oral squamous cell carcinoma (OSCC) is one of the most common malignancies in the world, with 50 000 new cases each year (1). Despite advancements in treatments over the past few decades, the 5-year survival rate of OSCC is still 0.05). Table 3 listed the genotype and allele frequencies of individual SNP in case and control subjects. We noticed that two SNPs, rs1130214 and rs3803300, were significantly associated with risk of OSCC. Compared with those carrying wild-type genotype and allele, patients carrying variant GT/TT genotype and T allele of rs1130214 had an increased risk of OSCC (OR = 1.966, 95% CI = 1.210–3.194; OR = 1.698, 95% CI = 1.086–2.656). As for rs3803300, variant genotypes AG and GG, especially the GG genotype, showed a strong association with higher OSCC susceptibility than the wild-type genotype AA (OR = 1.587, 95% CI = 1.036–2.432; OR = 2.948, 95% CI = 1.432–6.069). The minor G allele of rs3803300 also exhibited same association as AG/GG genotype (OR = 1.603, 95% CI = 1.190–2.160). However, there was no statistical difference in genotype and allele frequencies between cases and controls in the other three SNPs. Haplotype analysis Among the five polymorphisms, rs1130214 and rs3803300 were detected to be in strong Linkage disequilibrium (LD) with D’ = 0.99. Thus, we assessed the possible association Table 2 General characteristics for the case and control subjects
Characteristics
Levels
Age
Mean (SD) Male Female I II III IV
Gender Clinical stage
Cases (%) (N = 182)
Controls (%) (N = 207)
Pvalue
57.38 (11.791)
55.46 (10.658)
0.093
117 (56.52) 90 (43.48)
0.502
109 73 45 60 42 35
(59.89) (40.11) (24.73) (32.97) (23.08) (19.22)
3
Discussion AKT1, also known as PKBa, is a member of the AKT serine–threonine kinase family located on chromosome 14q32.32. AKT1 is a critical downstream effector of PI3K signal transduction pathway. Aberrant expression and genetic mutations in AKT1 affect a variety of cellular processes, including cell motility, proliferation, cell growth, and survival. These processes are critical to human cancer development, thus suggesting a role in OSCC (24, 25). Polymorphisms on AKT1 are associated with many types of human cancers. Rs2498801, a polymorphism on AKT1, was suggested to increase the risk of endometrial cancer in nonHispanic whites, African Americans, and Mexican Americans (26). Another two polymorphisms, rs2494738 and rs1130214, were found to be related to a more aggressive prostate cancer in Caucasian population (27). Meanwhile, the AA genotype of rs1130233 on AKT1 gene was reported to be associated with shorter time to progression and overall survival by affecting the mRNA expression level of AKT1 in non-small cell lung cancer patients (28). However, until now, the correlation between SNPs on AKT1 gene and OSCC has been unclear. This study is the first report about SNPs on AKT1 gene in OSCC. We investigated five AKT1 polymorphisms in a total of 207 cancer-free controls and 182 OSCC patients. All these SNPs have been extensively investigated in other human diseases before. The genotype and allele distribution of polymorphisms rs1130214 and rs3803300 were significantly different between these two groups, indicating that these two SNPs might be related to OSCC risk. In the current study, variant genotype and allele of rs1130214 was related to increased risk of OSCC, which was in accordance with the findings in non-small cell lung cancer and prostate J Oral Pathol Med
AKT1 gene SNPs in OSCC Wang et al.
4
Table 3 Association between selected SNP and OSCC susceptibility Cases (N = 182) (%)
Polymorphisms rs1130214 GG GT/TT G T rs1130233 GG GA AA G A rs2494732 CC CT TT C T rs3730358 CC CT/TT C T rs3803300 AA AG GG A G
Controls (N = 207) (%)
OR (95% CI)
P-value
130 52 312 52
(71.43) (28.57) (85.71) (14.29)
172 35 377 37
(83.09) (16.91) (91.06) (8.94)
1.000 1.966 (1.210–3.194) 1.000 1.698 (1.086–2.656)
1 ref 0.006 1 ref 0.019
28 95 59 151 213
(15.38) (52.20) (32.42) (41.48) (58.52)
35 93 79 163 251
(16.91) (44.93) (38.16) (39.37) (60.63)
1.000 1.277 (0.720–2.265) 0.934 (0.512–1.702) 1.000 0.916 (0.688–1.221)
1 ref 0.403 0.822 1 ref 0.549
97 71 14 265 99
(53.30) (39.01) (7.69) (72.80) (27.20)
111 80 16 302 112
(53.62) (38.65) (7.73) (72.95) (27.05)
1.000 1.016 (0.667–1.546) 1.001 (0.465–2.157) 1.000 1.007 (0.734–1.383)
1 ref 0.942 0.997 1 ref 0.964
166 16 348 16
(91.21) (8.79) (95.60) (4.40)
188 19 394 20
(90.82) (9.18) (95.17) (4.83)
1.000 0.954 (0.475–1.915) 1.000 0.906 (0.462–1.775)
1 ref 0.894 1 ref 0.773
63 93 26 219 145
(34.62) (51.10) (14.2) (60.16) (39.84)
100 93 14 293 121
(48.31) (44.93) (6.76) (70.77) (29.23)
1.000 1.587 (1.036–2.432) 2.948 (1.432–6.069) 1.000 1.603 (1.190–2.160)
1 ref 0.033 0.003 1 ref 0.002
OR, odds ratio; CI, confidence interval. The bold numbers mean the P-value is
J Oral Pathol Med © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd wileyonlinelibrary.com/journal/jop
Genetic variants in AKT1 gene were associated with risk and survival of OSCC in Chinese Han Population Yun Wang1,†, Lin Lin1,†, Hao Xu1,2, Taiwen Li1, Yu Zhou1, Hongxia Dan1, Lu Jiang1, Ga Liao1, Min Zhou1, Longjiang Li1, Xin Zeng1, Jing Li1, Qianming Chen1 1
State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China; 2West China School of Public Health, Sichuan University, Chengdu, China
BACKGROUND: AKT1 is an important downstream effector of PTEN/PI3K/AKT signal transduction pathway. Aberrant expression and genetic variant of AKT1 gene are suggested to be involved in several types of human cancers, including OSCC. The aim of this study was to investigate the possible association between AKT1 gene polymorphisms and OSCC in Chinese Han Population. METHODS: A total of 182 OSCC patients and 207 cancer-free controls were enrolled for this hospital-based study. Five single-nucleotide polymorphisms (SNPs) on AKT1 (rs1130214, rs1130233, rs2494732, rs3730358, rs3803300) were investigated and genotyped by Sequenom Mass ARRAY & iPLEX-MALDI-TOF technology. Chi-square test, SHEsis software, and Kaplan–Meier method were used to evaluate the relationship between selected SNPs and OSCC susceptibility and progression. RESULTS: Significant difference of genotype distribution was observed between cases and control group at SNP sites rs1130214 (P = 0.006) and rs3803300 (P = 0.033, P = 0.003 for heterozygote and homozygous mutant, respectively). In the haplotype analysis, haplotype H4 which contained mutant-type allele of rs1130214 and rs3803300 was also related to OSCC risk (OR = 1.974, 95% CI = 1.048–3.718). Moreover, CT genotype of rs3730358 was associated with higher risk of OSCC progression (HR = 2.466, 95% CI = 1.017–5.981). CONCLUSION: Our results indicated that rs1130214 and rs3803300 were related to OSCC susceptibility in Chinese Han Population. In addition, rs3730358 might be associated with progression-free survival time of OSCC patients, suggesting that this SNP could be a potential prognosis marker for OSCC. J Oral Pathol Med (2014)
Correspondence: Jing Li and Xin Zeng, State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, No.14, Section 3, Renmin Nan Road, Chengdu 610041, China. Tel: +8615928152276, Fax: +86-02885501484, E-mails: [email protected] and [email protected] † These authors contributed equally to this paper. Accepted for publication April 28, 2014
Keywords: AKT1; oral squamous cell carcinoma; single-nucleotide polymorphism; survival; susceptibility
Introduction Oral squamous cell carcinoma (OSCC) is one of the most common malignancies in the world, with 50 000 new cases each year (1). Despite advancements in treatments over the past few decades, the 5-year survival rate of OSCC is still 0.05). Table 3 listed the genotype and allele frequencies of individual SNP in case and control subjects. We noticed that two SNPs, rs1130214 and rs3803300, were significantly associated with risk of OSCC. Compared with those carrying wild-type genotype and allele, patients carrying variant GT/TT genotype and T allele of rs1130214 had an increased risk of OSCC (OR = 1.966, 95% CI = 1.210–3.194; OR = 1.698, 95% CI = 1.086–2.656). As for rs3803300, variant genotypes AG and GG, especially the GG genotype, showed a strong association with higher OSCC susceptibility than the wild-type genotype AA (OR = 1.587, 95% CI = 1.036–2.432; OR = 2.948, 95% CI = 1.432–6.069). The minor G allele of rs3803300 also exhibited same association as AG/GG genotype (OR = 1.603, 95% CI = 1.190–2.160). However, there was no statistical difference in genotype and allele frequencies between cases and controls in the other three SNPs. Haplotype analysis Among the five polymorphisms, rs1130214 and rs3803300 were detected to be in strong Linkage disequilibrium (LD) with D’ = 0.99. Thus, we assessed the possible association Table 2 General characteristics for the case and control subjects
Characteristics
Levels
Age
Mean (SD) Male Female I II III IV
Gender Clinical stage
Cases (%) (N = 182)
Controls (%) (N = 207)
Pvalue
57.38 (11.791)
55.46 (10.658)
0.093
117 (56.52) 90 (43.48)
0.502
109 73 45 60 42 35
(59.89) (40.11) (24.73) (32.97) (23.08) (19.22)
3
Discussion AKT1, also known as PKBa, is a member of the AKT serine–threonine kinase family located on chromosome 14q32.32. AKT1 is a critical downstream effector of PI3K signal transduction pathway. Aberrant expression and genetic mutations in AKT1 affect a variety of cellular processes, including cell motility, proliferation, cell growth, and survival. These processes are critical to human cancer development, thus suggesting a role in OSCC (24, 25). Polymorphisms on AKT1 are associated with many types of human cancers. Rs2498801, a polymorphism on AKT1, was suggested to increase the risk of endometrial cancer in nonHispanic whites, African Americans, and Mexican Americans (26). Another two polymorphisms, rs2494738 and rs1130214, were found to be related to a more aggressive prostate cancer in Caucasian population (27). Meanwhile, the AA genotype of rs1130233 on AKT1 gene was reported to be associated with shorter time to progression and overall survival by affecting the mRNA expression level of AKT1 in non-small cell lung cancer patients (28). However, until now, the correlation between SNPs on AKT1 gene and OSCC has been unclear. This study is the first report about SNPs on AKT1 gene in OSCC. We investigated five AKT1 polymorphisms in a total of 207 cancer-free controls and 182 OSCC patients. All these SNPs have been extensively investigated in other human diseases before. The genotype and allele distribution of polymorphisms rs1130214 and rs3803300 were significantly different between these two groups, indicating that these two SNPs might be related to OSCC risk. In the current study, variant genotype and allele of rs1130214 was related to increased risk of OSCC, which was in accordance with the findings in non-small cell lung cancer and prostate J Oral Pathol Med
AKT1 gene SNPs in OSCC Wang et al.
4
Table 3 Association between selected SNP and OSCC susceptibility Cases (N = 182) (%)
Polymorphisms rs1130214 GG GT/TT G T rs1130233 GG GA AA G A rs2494732 CC CT TT C T rs3730358 CC CT/TT C T rs3803300 AA AG GG A G
Controls (N = 207) (%)
OR (95% CI)
P-value
130 52 312 52
(71.43) (28.57) (85.71) (14.29)
172 35 377 37
(83.09) (16.91) (91.06) (8.94)
1.000 1.966 (1.210–3.194) 1.000 1.698 (1.086–2.656)
1 ref 0.006 1 ref 0.019
28 95 59 151 213
(15.38) (52.20) (32.42) (41.48) (58.52)
35 93 79 163 251
(16.91) (44.93) (38.16) (39.37) (60.63)
1.000 1.277 (0.720–2.265) 0.934 (0.512–1.702) 1.000 0.916 (0.688–1.221)
1 ref 0.403 0.822 1 ref 0.549
97 71 14 265 99
(53.30) (39.01) (7.69) (72.80) (27.20)
111 80 16 302 112
(53.62) (38.65) (7.73) (72.95) (27.05)
1.000 1.016 (0.667–1.546) 1.001 (0.465–2.157) 1.000 1.007 (0.734–1.383)
1 ref 0.942 0.997 1 ref 0.964
166 16 348 16
(91.21) (8.79) (95.60) (4.40)
188 19 394 20
(90.82) (9.18) (95.17) (4.83)
1.000 0.954 (0.475–1.915) 1.000 0.906 (0.462–1.775)
1 ref 0.894 1 ref 0.773
63 93 26 219 145
(34.62) (51.10) (14.2) (60.16) (39.84)
100 93 14 293 121
(48.31) (44.93) (6.76) (70.77) (29.23)
1.000 1.587 (1.036–2.432) 2.948 (1.432–6.069) 1.000 1.603 (1.190–2.160)
1 ref 0.033 0.003 1 ref 0.002
OR, odds ratio; CI, confidence interval. The bold numbers mean the P-value is
