P73和MDM2基因多态性与卵巢上皮性癌发病风险的关联研究
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摘要
目的:卵巢癌是妇科恶性肿瘤死亡的最常见的病因,其中90%是卵巢上皮性癌。卵巢上皮性癌因发病隐匿,进展迅速,约有70%的患者发现时已属晚期,5年生存率仅为25%～30%。有研究显示,遗传易感性可能在卵巢癌的发生中起重要作用。因此,寻找易感基因,确立高危人群,实现早期诊断是提高卵巢癌生存率的有效途径。近年来,p73和MDM2(murine double minute 2)作为肿瘤候选基因,已成为人们研究的热点。
p73是一种候选的抑癌基因,在调控细胞周期和维持细胞稳定等方面具有重要作用,其第二外显子非编码区存在两个单核苷酸多态性(single nucleotide polymorphisms, SNPs)(G4C14/A4T14),可以形成茎环结构而影响基因表达。而MDM2是一种原癌基因,也是p73重要的负调节因子,通过抑制p73的转录活性进而减弱其功能。鉴于p73、MDM2在细胞周期控制和凋亡中的相互作用,本研究旨在探讨p73和MDM2基因单核苷酸多态性(SNPs)与卵巢上皮性癌发病风险的关系,从分子水平为卵巢癌的预防和诊治提供实验依据。
方法:采用基于医院的病例-对照研究方法,以257例卵巢上皮癌患者和257例健康志愿者妇女(对照组)为研究对象,采集所有研究对象的外周静脉血5 ml,以枸橼酸钠抗凝,采用蛋白酶K消化-饱和氯化钠盐析法提取白细胞DNA。采用聚合酶链反应-限制性片段长度多态性( polymerase chain reaction-restriction fragment length polymorphism, PCR-RFLP )方法对p73 G4C14/A4T14、MDM2 309T/G和MDM2 Del1518+/-这3个SNPs位点进行基因分型。
数据分析采用SPSS 11.5统计软件进行处理。对照组基因型频率分布行χ2检验做Hardy-Weinberg平衡分析。病例组与对照组的年龄差异采用t检验,基因型和等位基因频率分布比较采用χ2检验,以非条件logistic回归方法计算相对风险度的比值比(odds ratio, OR)及其95%可信区间(confidence interval, CI)。采用EH软件分析MDM2 309T/G和MDM2Del1518+/-基因单体型频率,2ld软件分析连锁不平衡状态。采用似然比检验(likelihood ratio test)分析p73和MDM2基因之间的交互作用。以P<0.05为差异有统计学意义。
结果:
1对照组p73 G4C14/A4T14和MDM2 309T/G,Del1518+/-多态的基因型分布均符合Hardy-Weinberg平衡(χ2=0.00, 1.00和0.64,所有P值>0.05)。
2卵巢癌组和对照组中,p73 G4C14/A4T14多态的基因型和等位基因分布频率的差异无统计学意义(P=0.20和P=0.55)。与GC/AT+AT/AT基因型比较,GC/GC基因型与卵巢癌的发病风险无关,OR值为1.24 (95%CI=0.87～1.77)。以病理类型、临床分期或确诊时年龄分层分析,p73 G4C14/A4T14多态的基因型频率在各组间的分布差异也无统计学意义(P值均大于0.05)。
3 MDM2 309T/G的3种基因型频率(T/T、T/G、G/G)在卵巢癌组和对照组中分别是30.0%、46.7%、23.3%和21.8%、47.1%、31.1%,两组比较差异有统计学意义(P=0.046)。卵巢癌组中的MDM2 309T/G SNP的G等位基因频率(46.7%)明显低于对照组(54.7%),两组比较差异有统计学意义(P=0.01)。与T/T基因型相比,携带G等位基因型(T/G+G/G基因型)可显著降低卵巢上皮癌的发病风险(OR=0.65, 95%CI=0.44～0.97)。以病理类型,临床分期和确诊年龄分层分析发现,携带G等位基因型(T/G+G/G基因型)可明显降低内膜样癌,晚期卵巢癌和50岁以上的卵巢癌病人的发病风险( OR=0.53, 95%CI=0.31～0.90; OR=0.62, 95%CI=0.40～0.97; OR=0.59, 95%CI=0.38～0.92)。
4 MDM2 Del1518+/- SNP的基因型和等位基因频率在病例组和对照组中的分布差异无统计学意义(P=0.68和P=0.45)。与(+/-)+(-/-)基因型相比,+/+基因型与卵巢上皮癌的发病风险无关(OR=1.17, 95%CI=0.83～1.65)。但以病理类型、临床分期和发病年龄分层分析显示,携带MDM2 Del1518+/+基因型有增加黏液性卵巢癌和早期卵巢癌发病风险的趋势(OR=2.01, 95%CI=0.93～4.37; OR=1.64, 95%CI =0.99～2.72),而与发病年龄无明显关联。
5采用2ld软件对MDM2 309T/G和Del1518+/-两个多态位点进行联合分析,结果显示两个位点间存在部分连锁不平衡现象(D’=0.57, P=0.00)。而采用EH软件对MDM2 309T/G和Del1518+/-两位点SNPs进行单体型分析显示,虽然4种单体型(+1518/T309, +1518/G309, -1518/T309和-1518/G309)频率在卵巢癌组与对照组中的分布差异无统计学意义(P=0.07),但与+1518/T309单体型相比,+1518/G309和-1518/G309单体型均可明显降低卵巢上皮性癌的发病风险(OR=0.73, 95%CI=0.55～0.98; OR=0.65, 95%CI=0.45～0.96)。
6采用似然比检验分析p73 G4C14/A4T14 SNP与MDM2 309T/G、Del1518+/- 2个SNPs之间的基因-基因交互作用。结果发现,p73 G4C14/A4T14与MDM2 309T/G之间有明显的交互作用(P=0.03, OR=0.89, 95%CI=0.80～0.99);而p73 G4C14/A4T14与Del1518+/-之间无明显交互作用(P=0.36)。与GC/GCp73 G4C14/A4T14-T/TMDM2 309T/G基因型相比,携带(GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G基因型可明显降低卵巢上皮性癌的发病风险(OR=0.55,95%CI=0.31～0.98)。
结论:
1 p73 G4C14/A4T14的基因多态性可能与卵巢上皮性癌遗传易感性无关。
2携带MDM2 309T/G多态的G等位基因型可能显著降低卵巢上皮性癌的发病风险,并与卵巢上皮性癌的病理类型、临床分期及确诊年龄有显著关联,因此,309G等位基因可能是卵巢上皮性癌发病的一个潜在的保护因素。
3 MDM2 Del1518+/+基因型有增加黏液性卵巢癌和早期卵巢癌发病风险的趋势,而与发病年龄无明显关联。
4 MDM2 309T/G和Del1518+/-两个多态位点间存在部分连锁不平衡现象,+1518/G309和-1518/G309单体型均可明显降低卵巢上皮性癌的发病风险。
5 p73 G4C14/A4T14与MDM2 309T/G之间有明显交互作用,而p73 G4C14/A4T14与Del1518+/-之间无明显交互作用存在。(GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G基因型可明显降低卵巢上皮性癌的发病风险。
Objective: Ovarian cancer (OC) is the most common cause of death of the gynecological malignancies in Chinese women. Epithelial ovarian cancer (EOC) originates from ovarian surface epithelium and occupies 90% of all ovarian tumors in women. Nevertheless, it is often asymptomatic and approximately 70% of all cases, when diagnosed, present an advanced stage of the disease. So the overall 5-year survival rate is only 25 to 30% currently. The study showed that genetic susceptibility that affect the function of pathways known to in?uence the neoplastic process may be particularly relevant. Therefor, it is the effective way to improve the survival rate of ovarian cancer to look for predisposing genes, high-risk group and early diagnosis method. p73 and murine double minute2 (MDM2), as the candidate tumor gene, have been the studyed focus in recent years.
p73, as a candidate tumour suppressor gene, plays an important role in modulating cell-cycle control and sustaining cell stabilization. A dinucleotide single nucleotide polymorphisms (SNPs) at positions 4 and 14 of exon 2 (G4C14/A4T14) is regarded as a functional one, which possibly forms a stem-loop structure to influence the p73 translation. MDM2, as a proto-oncogene, is an important negative regulator of p73, which represses the transcriptional activity of p73 and thus attenuates its function. Based on the molecular interaction between p73 and MDM2 in cell-cycle control and apoptosis, this study was designed to investigate the association between single nucleotide polymorphisms (SNPs) in the promoter region of p73 and MDM2 genes and the risk of epithelial ovarian cancer. By this way, we hope to offer some evidences for the prevention and therapy of ovarian cancer at molecular level.
Methods: This hospital-based case-control study included 257 patients with epithelial ovarian cancer and 257 healthy women as control. Five ml of venous blood from each subject was drawn in vacutainer tubes. The white blood cell DNA was extracted by using proteinase K-digestion followed by a salting out procedure. p73 G4C14/A4T14 and MDM2 309T/G and MDM2 Del1518+/- SNPs were genotyped by polymerase chain reaction-restrictive fragment length polymorphism (PCR-RFLP) analysis.
Statistical analysis was performed using SPSS11.5 software package. Hardy-Weinberg analysis was performed by comparing the observed and expected genotype frequencies in study groups using Chi-square test. Between cases and controls, the t test was used to examine the difference of ages and ages of menarche and the Chi-square test was used to examine the difference of gravidity and parity. Univariate comparisons of allele and genotype distribution were performed using Chi-square test. The odds ratio (OR) and 95% confidence interval (CI) were calculated using an unconditional logistic regression model. The MDM2 309T/G and MDM2 Del1518+/- haplotype frequencies and linkage disequilibrium coefficient were estimated by using EH linkage software and 2ld software. Tests for the interaction between p73 and MDM2 genes were performed using the likelihood ratio test. P<0.05 was considered significant for all statistical analyses.
Results:
1 The genotype distributions of the p73 G4C14/A4T14 and MDM2 309T/G, Del1518+/- SNPs in control group did not significantly deviate from that expected for a Hardy-Weinberg equilibrium (χ2=0.00, 1.00 and 0.64, respectively, and all P values >0.05).
2 There were no significant differences in allele frequencies and genotype distributions of the p73 G4C14/A4T14 polymorphism between cases and control women (P=0.55 and 0.20, respectively). Compared to the GC/AT+AT/AT genotypes, the GC/GC genotype had no association with the risk of epithelial ovarian cancer (OR=1.24, 95%CI=0.87～1.77). Stratification analysis showed no significant difference between the cases and control groups in allele or genotype distributions of p73 G4C14/A4T14 according to the pathological type, clinical stage and patient age of epithelial ovarian cancer (P>0.05).
3 The frequencies of three genotypes (T/T, T/G and G/G) of MDM2 309T/G in ovarian cancer cases and controls were 30.0%, 46.7%, 23.3% and 21.8%, 47.1%, 31.1%,respectively, and there was significant difference between the two groups (P=0.046). The frequencies of the G allele of the MDM2 309T/G SNP were significantly lower in ovarian cancer cases (46.7%) than those in healthy controls (54.7%), there was statistical difference between the two groups (P=0.01). Compared with the T/T genotype, the G allelotype (T/G+G/G genotype) significantly decreased the risk of developing EOC (OR=0.65, 95%CI=0.44～0.97). Stratification analysis showed that subjects carrying G allelotype (T/G+G/G genotype) significantly decreased the risk of endometrioid ovarian cancer (OR=0.53, 95%CI=0.31～0.90) according to histological subtypes. When stratified by clinical stage, individuals with the G allelotype (T/G+G/G genotype) significantly had a relation on the reduced risk of the advanced ovarian cancer (OR=0.62,95%CI=0.40～0.97). Furthermore, the lower risk of ovarian cancer was associated with the G allelotype (T/G+G/G genotype) in subjects that were 50 or older, with odds ratio of 0.59 (95%CI=0.38～0.92).
4 MDM2 Del1518+/- genotype and allele frequencies did not differ between the case group and the control group (P=0.68 and P=0.45). Compared to the (+/-)+(-/-) genotypes, the +/+ genotype did not significantly modify the risk of epithelial ovarian cancer with odds ratio of 1.17 (95%CI=0.83～1.65). But MDM2 Del1518+/+ genotype was associated with a trend for mucinous ovarian cancer by stratification analysis according to histological subtypes (OR=2.01, 95%CI=0.93～4.37). When stratified by clinical stage, individuals with the +/+ genotype tended to increased the risk of the earlier ovarian cancer (OR=1.64, 95%CI=0.99～2.72). No association was observed between the polymorphism and age at diagnosis.
5 The results of the 2LD program analysis showed that the MDM2 309T/G and Del1518+/- polymorphisms displayed linkage disequilibrium (D’=0.57, P=0.00). The frequencies of four haplotypes (+1518/T309, +1518/G309, -1518/T309 and -1518/G309) of MDM2 309T/G and Del1518+/- were not significantly different between the case and control groups (P=0.07). Compared with the haplotype of +1518/T309, the +1518/G309 and -1518/G309 haplotypes both significantly decreased the risk of EOC (OR=0.73, 95%CI=0.55～0.98; OR=0.65, 95%CI=0.45～0.96).
6 The gene-gene interaction between the p73 G4C14/A4T14 and MDM2 309T/G or Del1518+/- SNPs on the risk of EOC was analyzed using the likelihood ratio test. The results showed that there was a significant interaction between p73 G4C14/A4T14 and MDM2 309T/G (P=0.03, OR=0.89, 95%CI=0.80～0.99). However, the significant interaction of p73 G4C14- to-A4T14 and Del1518+/- was not observed (P=0.36). Compared with GC/GCp73 G4C14/A4T14-T/TMDM2 309T/G, subjects carrying (GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G significantly decreased the risk of EOC (OR=0.55, 95%CI=0.31～0.98).
Conclusions:
1 The p73 G4C14/A4T14 SNP may have no susceptibility to the epithelial ovarian cancer.
2 MDM2 309G allele carriers significantly decreased the risk of epithelial ovarian cancer. Furthermore, there were significant association between MDM2 309T/G polymorphism and tumor histological subtypes, clinical stage and age. It may be a potentially protective factor for epithelial ovarian cancer.
3 The MDM2 Del1518+/+ genotype tended to increase the risk of mucinous ovarian cancer or earlier ovarian cancer, but not associated with the age.
4 The MDM2 309T/G and Del1518+/- SNPs showed linkage disequilibrium, the +1518/G309 and -1518/G309 haplotypes both significantly decreased the risk of EOC.
5 There was a significant interaction between p73 G4C14/A4T14 and MDM2 309T/G. But the significant interaction of p73 G4C14/A4T14 and Del1518+/- was not observed. (GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G genetype significantly decreased the risk of EOC.
p73是一种候选的抑癌基因,在调控细胞周期和维持细胞稳定等方面具有重要作用,其第二外显子非编码区存在两个单核苷酸多态性(single nucleotide polymorphisms, SNPs)(G4C14/A4T14),可以形成茎环结构而影响基因表达。而MDM2是一种原癌基因,也是p73重要的负调节因子,通过抑制p73的转录活性进而减弱其功能。鉴于p73、MDM2在细胞周期控制和凋亡中的相互作用,本研究旨在探讨p73和MDM2基因单核苷酸多态性(SNPs)与卵巢上皮性癌发病风险的关系,从分子水平为卵巢癌的预防和诊治提供实验依据。
方法:采用基于医院的病例-对照研究方法,以257例卵巢上皮癌患者和257例健康志愿者妇女(对照组)为研究对象,采集所有研究对象的外周静脉血5 ml,以枸橼酸钠抗凝,采用蛋白酶K消化-饱和氯化钠盐析法提取白细胞DNA。采用聚合酶链反应-限制性片段长度多态性( polymerase chain reaction-restriction fragment length polymorphism, PCR-RFLP )方法对p73 G4C14/A4T14、MDM2 309T/G和MDM2 Del1518+/-这3个SNPs位点进行基因分型。
数据分析采用SPSS 11.5统计软件进行处理。对照组基因型频率分布行χ2检验做Hardy-Weinberg平衡分析。病例组与对照组的年龄差异采用t检验,基因型和等位基因频率分布比较采用χ2检验,以非条件logistic回归方法计算相对风险度的比值比(odds ratio, OR)及其95%可信区间(confidence interval, CI)。采用EH软件分析MDM2 309T/G和MDM2Del1518+/-基因单体型频率,2ld软件分析连锁不平衡状态。采用似然比检验(likelihood ratio test)分析p73和MDM2基因之间的交互作用。以P<0.05为差异有统计学意义。
结果:
1对照组p73 G4C14/A4T14和MDM2 309T/G,Del1518+/-多态的基因型分布均符合Hardy-Weinberg平衡(χ2=0.00, 1.00和0.64,所有P值>0.05)。
2卵巢癌组和对照组中,p73 G4C14/A4T14多态的基因型和等位基因分布频率的差异无统计学意义(P=0.20和P=0.55)。与GC/AT+AT/AT基因型比较,GC/GC基因型与卵巢癌的发病风险无关,OR值为1.24 (95%CI=0.87～1.77)。以病理类型、临床分期或确诊时年龄分层分析,p73 G4C14/A4T14多态的基因型频率在各组间的分布差异也无统计学意义(P值均大于0.05)。
3 MDM2 309T/G的3种基因型频率(T/T、T/G、G/G)在卵巢癌组和对照组中分别是30.0%、46.7%、23.3%和21.8%、47.1%、31.1%,两组比较差异有统计学意义(P=0.046)。卵巢癌组中的MDM2 309T/G SNP的G等位基因频率(46.7%)明显低于对照组(54.7%),两组比较差异有统计学意义(P=0.01)。与T/T基因型相比,携带G等位基因型(T/G+G/G基因型)可显著降低卵巢上皮癌的发病风险(OR=0.65, 95%CI=0.44～0.97)。以病理类型,临床分期和确诊年龄分层分析发现,携带G等位基因型(T/G+G/G基因型)可明显降低内膜样癌,晚期卵巢癌和50岁以上的卵巢癌病人的发病风险( OR=0.53, 95%CI=0.31～0.90; OR=0.62, 95%CI=0.40～0.97; OR=0.59, 95%CI=0.38～0.92)。
4 MDM2 Del1518+/- SNP的基因型和等位基因频率在病例组和对照组中的分布差异无统计学意义(P=0.68和P=0.45)。与(+/-)+(-/-)基因型相比,+/+基因型与卵巢上皮癌的发病风险无关(OR=1.17, 95%CI=0.83～1.65)。但以病理类型、临床分期和发病年龄分层分析显示,携带MDM2 Del1518+/+基因型有增加黏液性卵巢癌和早期卵巢癌发病风险的趋势(OR=2.01, 95%CI=0.93～4.37; OR=1.64, 95%CI =0.99～2.72),而与发病年龄无明显关联。
5采用2ld软件对MDM2 309T/G和Del1518+/-两个多态位点进行联合分析,结果显示两个位点间存在部分连锁不平衡现象(D’=0.57, P=0.00)。而采用EH软件对MDM2 309T/G和Del1518+/-两位点SNPs进行单体型分析显示,虽然4种单体型(+1518/T309, +1518/G309, -1518/T309和-1518/G309)频率在卵巢癌组与对照组中的分布差异无统计学意义(P=0.07),但与+1518/T309单体型相比,+1518/G309和-1518/G309单体型均可明显降低卵巢上皮性癌的发病风险(OR=0.73, 95%CI=0.55～0.98; OR=0.65, 95%CI=0.45～0.96)。
6采用似然比检验分析p73 G4C14/A4T14 SNP与MDM2 309T/G、Del1518+/- 2个SNPs之间的基因-基因交互作用。结果发现,p73 G4C14/A4T14与MDM2 309T/G之间有明显的交互作用(P=0.03, OR=0.89, 95%CI=0.80～0.99);而p73 G4C14/A4T14与Del1518+/-之间无明显交互作用(P=0.36)。与GC/GCp73 G4C14/A4T14-T/TMDM2 309T/G基因型相比,携带(GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G基因型可明显降低卵巢上皮性癌的发病风险(OR=0.55,95%CI=0.31～0.98)。
结论:
1 p73 G4C14/A4T14的基因多态性可能与卵巢上皮性癌遗传易感性无关。
2携带MDM2 309T/G多态的G等位基因型可能显著降低卵巢上皮性癌的发病风险,并与卵巢上皮性癌的病理类型、临床分期及确诊年龄有显著关联,因此,309G等位基因可能是卵巢上皮性癌发病的一个潜在的保护因素。
3 MDM2 Del1518+/+基因型有增加黏液性卵巢癌和早期卵巢癌发病风险的趋势,而与发病年龄无明显关联。
4 MDM2 309T/G和Del1518+/-两个多态位点间存在部分连锁不平衡现象,+1518/G309和-1518/G309单体型均可明显降低卵巢上皮性癌的发病风险。
5 p73 G4C14/A4T14与MDM2 309T/G之间有明显交互作用,而p73 G4C14/A4T14与Del1518+/-之间无明显交互作用存在。(GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G基因型可明显降低卵巢上皮性癌的发病风险。
Objective: Ovarian cancer (OC) is the most common cause of death of the gynecological malignancies in Chinese women. Epithelial ovarian cancer (EOC) originates from ovarian surface epithelium and occupies 90% of all ovarian tumors in women. Nevertheless, it is often asymptomatic and approximately 70% of all cases, when diagnosed, present an advanced stage of the disease. So the overall 5-year survival rate is only 25 to 30% currently. The study showed that genetic susceptibility that affect the function of pathways known to in?uence the neoplastic process may be particularly relevant. Therefor, it is the effective way to improve the survival rate of ovarian cancer to look for predisposing genes, high-risk group and early diagnosis method. p73 and murine double minute2 (MDM2), as the candidate tumor gene, have been the studyed focus in recent years.
p73, as a candidate tumour suppressor gene, plays an important role in modulating cell-cycle control and sustaining cell stabilization. A dinucleotide single nucleotide polymorphisms (SNPs) at positions 4 and 14 of exon 2 (G4C14/A4T14) is regarded as a functional one, which possibly forms a stem-loop structure to influence the p73 translation. MDM2, as a proto-oncogene, is an important negative regulator of p73, which represses the transcriptional activity of p73 and thus attenuates its function. Based on the molecular interaction between p73 and MDM2 in cell-cycle control and apoptosis, this study was designed to investigate the association between single nucleotide polymorphisms (SNPs) in the promoter region of p73 and MDM2 genes and the risk of epithelial ovarian cancer. By this way, we hope to offer some evidences for the prevention and therapy of ovarian cancer at molecular level.
Methods: This hospital-based case-control study included 257 patients with epithelial ovarian cancer and 257 healthy women as control. Five ml of venous blood from each subject was drawn in vacutainer tubes. The white blood cell DNA was extracted by using proteinase K-digestion followed by a salting out procedure. p73 G4C14/A4T14 and MDM2 309T/G and MDM2 Del1518+/- SNPs were genotyped by polymerase chain reaction-restrictive fragment length polymorphism (PCR-RFLP) analysis.
Statistical analysis was performed using SPSS11.5 software package. Hardy-Weinberg analysis was performed by comparing the observed and expected genotype frequencies in study groups using Chi-square test. Between cases and controls, the t test was used to examine the difference of ages and ages of menarche and the Chi-square test was used to examine the difference of gravidity and parity. Univariate comparisons of allele and genotype distribution were performed using Chi-square test. The odds ratio (OR) and 95% confidence interval (CI) were calculated using an unconditional logistic regression model. The MDM2 309T/G and MDM2 Del1518+/- haplotype frequencies and linkage disequilibrium coefficient were estimated by using EH linkage software and 2ld software. Tests for the interaction between p73 and MDM2 genes were performed using the likelihood ratio test. P<0.05 was considered significant for all statistical analyses.
Results:
1 The genotype distributions of the p73 G4C14/A4T14 and MDM2 309T/G, Del1518+/- SNPs in control group did not significantly deviate from that expected for a Hardy-Weinberg equilibrium (χ2=0.00, 1.00 and 0.64, respectively, and all P values >0.05).
2 There were no significant differences in allele frequencies and genotype distributions of the p73 G4C14/A4T14 polymorphism between cases and control women (P=0.55 and 0.20, respectively). Compared to the GC/AT+AT/AT genotypes, the GC/GC genotype had no association with the risk of epithelial ovarian cancer (OR=1.24, 95%CI=0.87～1.77). Stratification analysis showed no significant difference between the cases and control groups in allele or genotype distributions of p73 G4C14/A4T14 according to the pathological type, clinical stage and patient age of epithelial ovarian cancer (P>0.05).
3 The frequencies of three genotypes (T/T, T/G and G/G) of MDM2 309T/G in ovarian cancer cases and controls were 30.0%, 46.7%, 23.3% and 21.8%, 47.1%, 31.1%,respectively, and there was significant difference between the two groups (P=0.046). The frequencies of the G allele of the MDM2 309T/G SNP were significantly lower in ovarian cancer cases (46.7%) than those in healthy controls (54.7%), there was statistical difference between the two groups (P=0.01). Compared with the T/T genotype, the G allelotype (T/G+G/G genotype) significantly decreased the risk of developing EOC (OR=0.65, 95%CI=0.44～0.97). Stratification analysis showed that subjects carrying G allelotype (T/G+G/G genotype) significantly decreased the risk of endometrioid ovarian cancer (OR=0.53, 95%CI=0.31～0.90) according to histological subtypes. When stratified by clinical stage, individuals with the G allelotype (T/G+G/G genotype) significantly had a relation on the reduced risk of the advanced ovarian cancer (OR=0.62,95%CI=0.40～0.97). Furthermore, the lower risk of ovarian cancer was associated with the G allelotype (T/G+G/G genotype) in subjects that were 50 or older, with odds ratio of 0.59 (95%CI=0.38～0.92).
4 MDM2 Del1518+/- genotype and allele frequencies did not differ between the case group and the control group (P=0.68 and P=0.45). Compared to the (+/-)+(-/-) genotypes, the +/+ genotype did not significantly modify the risk of epithelial ovarian cancer with odds ratio of 1.17 (95%CI=0.83～1.65). But MDM2 Del1518+/+ genotype was associated with a trend for mucinous ovarian cancer by stratification analysis according to histological subtypes (OR=2.01, 95%CI=0.93～4.37). When stratified by clinical stage, individuals with the +/+ genotype tended to increased the risk of the earlier ovarian cancer (OR=1.64, 95%CI=0.99～2.72). No association was observed between the polymorphism and age at diagnosis.
5 The results of the 2LD program analysis showed that the MDM2 309T/G and Del1518+/- polymorphisms displayed linkage disequilibrium (D’=0.57, P=0.00). The frequencies of four haplotypes (+1518/T309, +1518/G309, -1518/T309 and -1518/G309) of MDM2 309T/G and Del1518+/- were not significantly different between the case and control groups (P=0.07). Compared with the haplotype of +1518/T309, the +1518/G309 and -1518/G309 haplotypes both significantly decreased the risk of EOC (OR=0.73, 95%CI=0.55～0.98; OR=0.65, 95%CI=0.45～0.96).
6 The gene-gene interaction between the p73 G4C14/A4T14 and MDM2 309T/G or Del1518+/- SNPs on the risk of EOC was analyzed using the likelihood ratio test. The results showed that there was a significant interaction between p73 G4C14/A4T14 and MDM2 309T/G (P=0.03, OR=0.89, 95%CI=0.80～0.99). However, the significant interaction of p73 G4C14- to-A4T14 and Del1518+/- was not observed (P=0.36). Compared with GC/GCp73 G4C14/A4T14-T/TMDM2 309T/G, subjects carrying (GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G significantly decreased the risk of EOC (OR=0.55, 95%CI=0.31～0.98).
Conclusions:
1 The p73 G4C14/A4T14 SNP may have no susceptibility to the epithelial ovarian cancer.
2 MDM2 309G allele carriers significantly decreased the risk of epithelial ovarian cancer. Furthermore, there were significant association between MDM2 309T/G polymorphism and tumor histological subtypes, clinical stage and age. It may be a potentially protective factor for epithelial ovarian cancer.
3 The MDM2 Del1518+/+ genotype tended to increase the risk of mucinous ovarian cancer or earlier ovarian cancer, but not associated with the age.
4 The MDM2 309T/G and Del1518+/- SNPs showed linkage disequilibrium, the +1518/G309 and -1518/G309 haplotypes both significantly decreased the risk of EOC.
5 There was a significant interaction between p73 G4C14/A4T14 and MDM2 309T/G. But the significant interaction of p73 G4C14/A4T14 and Del1518+/- was not observed. (GC/AT+AT/AT)p73 G4C14/A4T14-(T/G+G/G)MDM2 309T/G genetype significantly decreased the risk of EOC.
引文
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