修回日期: 2008-06-15
接受日期: 2008-06-17
在线出版日期: 2008-08-08
目的: 探讨CCND1基因A870G多态与上海地区人群结直肠癌(colorectal cancer, CRC)遗传易感性的关系.
方法: 采用TaqMan MGB探针实时定量PCR法, 检测104例CRC与205名对照人群CCND1 A870G基因型分布及差异.
结果: CRC组和对照组870A等位基因频率分别为60.1%和52.4%, A的CRC发病风险是G的1.40倍(95%CI: 0.99-1.97, P = 0.057). 与GG基因型相比, GA基因型的CRC风险增加至1.99倍(95%CI: 0.94-4.20, P = 0.070), 而AA基因型的CRC风险显著增加至2.46倍(95%CI: 1.09-5.56, P = 0.031). 将GA、AA基因型合并计算, 则其CRC发病风险与GG基因型相比呈相似的显著性增加(OR = 2.13, 95%CI: 1.03-4.39, P = 0.040).
结论: CCND1 870A增加CRC发病风险, 可作为这一地区CRC高危人群的筛选指标.
引文著录: 荆佳晨, 熊文坚, 金惠, 稽贝纳. CCND1基因A870G多态与结直肠癌遗传易感性的相关性. 世界华人消化杂志 2008; 16(22): 2553-2556
Revised: June 15, 2008
Accepted: June 17, 2008
Published online: August 8, 2008
AIM: To investigate the possible association between CCND1 gene A870G polymorphism and susceptibility to colorectal cancer (CRC) in Shanghai population.
METHODS: CCND1 gene A870G genotypes were determined using real-time PCR technology with TaqMan MGB probes in 104 CRC cases and 205 sex-, age-matched controls.
RESULTS: The frequencies of A and G alleles were 0.601 and 0.399 in CRC cases, and 0.524 and 0.476 in controls, respectively. The A allele was marginally associated with CRC (P = 0.057) and had an increased risk for CRC (95%CI: 0.99-1.97) as compared with the G allele. Compared with GG genotype, GA genotype had a 1.99-fold increased risk (95%CI: 0.94-4.20, P = 0.070), whereas AA genotype had a 2.46-fold increased risk (95%CI: 1.09-5.56, P = 0.031) for CRC. The A-allele carriers (GA or AA genotypes) had a significantly elevated CRC risk (OR = 2.13, 95%CI: 1.03-4.39, P = 0.040) when as compared with GG genotype.
CONCLUSION: The A allele of CCND1 A870G is associated with the occurrence of CRC, which may serve as an index for screening high-risk population in Shanghai.
- Citation: Jing JC, Xiong WJ, Jin H, Ji BN. Relationship between CCND1 gene A870G polymorphism and genetic susceptibility to colorectal cancer. Shijie Huaren Xiaohua Zazhi 2008; 16(22): 2553-2556
- URL: https://www.wjgnet.com/1009-3079/full/v16/i22/2553.htm
- DOI: https://dx.doi.org/10.11569/wcjd.v16.i22.2553
CCND1基因编码产物Cyclin D1是一种关键的细胞周期调控蛋白, 促进细胞周期由G1期向S期转化, 其异常表达与肿瘤发生发展密切相关[1]. 研究发现, 人群中CCND1基因第870位点存在A或G(A870G)单核苷酸多态性. 功能学研究显示, 这种多态性影响Cyclin D1 mRNA剪切转录子的种类及特性[2]. 近来有关CCND1 A870G与包括结直肠癌(colorectal cancer, CRC)在内的肿瘤遗传易感性关系的研究备受关注. 本研究初步探讨了CCND1 A870G与上海地区人群CRC发病风险的关系.
本院2003-2006年CRC住院患者104例, 均经组织病理学确诊为腺癌. 其中男54例, 女50例, 患者年龄32-83(平均60.7±13.3)岁. CRC组对象纳入标准包括: 上海籍; 汉族; 年龄>30岁; 术前未经放疗、化疗等治疗. 剔除标准为家族性腺瘤性息肉病和遗传性非腺瘤性结肠癌. 205名对照为同时期同一地区健康无血缘关系的上海籍汉族个体. 对照组与CRC组在性别和年龄(±2岁)方面保持均衡可比. 本研究经医院伦理委员会批准, 所有研究对象均同意并填写知情同意书.
按常规蛋白酶K、酚/氯仿法提取外周血白细胞基因组DNA. 采用TaqMan MGB探针实时定量PCR法检测CCND1 A870G基因型, 由上海基康生物技术有限公司完成. 引物和探针序列: 上游引物F, 5'-ACG CTT CCT CTC CAG AGT GAT C-3'; 下游引物R, 5'-AGG CTG CCT GGG ACA TCA-3'; 等位基因A特异性TaqMan探针, 5'-FAM-TGA CCC AGT AAG TGA G-MGB-3'; 等位基因G特异性TaqMan探针, 5'-TET-TGA CCC GGT AAG TGA-MGB-3'; 扩增片段长度63 bp. PCR扩增体系(25 µL)中含: 1 ×TaqMan PCR master mix(Applied Biosystems, Foster City, CA), 1 µL(50 mg/L)基因组DNA, 1 µmol/L上游引物, 1 µmol/L下游引物, 0.2 µmol/L FAM-标记探针, 0.2 µmol/L TET-标记探针. 扩增参数: 50℃预热2 min, 95℃预变性10 min, 然后95℃变性15 s, 62℃复性1 min, 共40个循环. 最后在ABI 7900上检测荧光信号. 基因型分析结果用Sequence Detector System 2.0软件(Applied Biosystems, Foster City, CA)自动生成(图1).
统计学处理 以非条件Logistic回归法计算表示相对危险度的比值比(OR)及其95%可信区间(CI). OR值均经性别、年龄、吸烟、饮酒和CRC家族史因素校正. 两组人群CRC混淆因素比较采用χ2检验. 以P<0.05作为有统计学意义的检验水准. 数据统计应用统计分析系统SPSS10.0进行.
两组人群的性别、年龄、吸烟和饮酒构成差异无显著意义(P>0.05). CRC组和对照组的CRC家族史阳性者分别占10.2%和2.9%, 两者比较P = 0.022(表1).
| 分组 | n | 性别 | 年龄(岁) | 吸烟 | 饮酒 | CRC家族史 | |||||
| 女 | 男 | ≤60 | ＞60 | 否 | 是 | 否 | 是 | 无 | 有 | ||
| 对照 | 205 | 98(47.8) | 107(52.2) | 99(48.3) | 106(51.7) | 151(73.7) | 54(26.3) | 168(82.0) | 37(18.0) | 184(89.8) | 21(10.2) |
| CRC | 104 | 50(48.1) | 54(51.9) | 50(48.1) | 54(51.9) | 75(72.1) | 29(27.9) | 79(76.0) | 25(24.0) | 101(97.1) | 3(2.9) |
| χ2 | 0.002 | 0.001 | 0.084 | 1.543 | 5.216 | ||||||
| P值 | 0.964 | 0.971 | 0.772 | 0.214 | 0.022 | ||||||
对照组GG、GA和AA基因型频率分别为20.0%、55.1%和24.9%, 符合Hardy-Weinberg遗传平衡定律(P= 0.162), 而CRC组的基因型频率为GG 10.6%、GA 58.7%和AA 30.8%. 据此计算, CRC组中A等位基因频率为60.1%, 高于对照组的52.4%, A的CRC发病风险是G的1.40倍(95% CI = 0.99-1.97, P = 0.057). 与GG基因型相比, GA基因型的CRC风险增加至1.99倍(95% CI = 0.94-4.20, P = 0.070), 而AA基因型的CRC风险显著增加至2.46倍(95% CI = 1.09-5.56, P = 0.031). 将GA、AA基因型合并计算, 则其CRC发病风险与GG基因型相比呈相似的显著性增加(OR = 2.13, 95% CI = 1.03-4.39, P = 0.040, 表2).
CCND1基因870G→A多态性位点位于该基因第四外显子的转录剪接供给区, 可导致转录剪接的方式发生改变, 最终产生半衰期更长的Cyclin D1蛋白质, 结果核内Cyclin D1积聚, 肿瘤发生风险随之增加[2]. 与此功能学研究相一致, 本研究结果显示870A等位基因增加CRC发病风险. 检索文献, 尚未见以我国大陆人群为对象的CCND1 A870G与CRC遗传易感性关系的研究报道.
有关CCND1 A870G与CRC发病风险关系的研究在其它人群中已有报道, 但结果不完全一致, 或提示870A增加CRC风险[3-5], 或提示870G增加CRC风险[6], 或提示二者无关[7-9]. 新近Tan et al[7]对CCND1 A870G与CRC风险关系的荟萃分析结果显示, 870A显著增加CRC发病风险, 这支持我们的研究结果. Le Marchand et al[4]以多种族人群为对象的研究结果显示, 870A增加夏威夷本地人和白人、但不增加日本裔夏威夷人的CRC风险, 这提示人群差异可能是造成上述研究结果差异的重要原因之一. 此外, Huanget al[10]报道870A特异性增加年轻男性CRC风险, 与之相反, Schernhammer et al[11]报道870A特异性增加女性CRC风险. Probst-Hensch et al[12]报道, 870A特异性增加低异硫氰酸盐饮食/高活性GST基因型者的CRC风险. 这些研究结果提示, 饮食、遗传和性别等也是影响A870G与CRC发病风险关系的重要因素, 可能部分解释了上述研究结果差异的原因.
总之, CCND1基因A870G多态是上海人群重要的CRC遗传易感分子, 可作为CRC高危人群的筛选指标.
有关CCND1基因与恶性肿瘤关系的研究广泛, 但目前对该基因多态性研究仍处于初步阶段, CCND1基因A870G多态性与结直肠癌相关性的报道缺乏一致性.
曹秀峰, 主任医师, 南京医科大学附属南京第一医院肿瘤中心; 李淑德, 主任医师, 中国人民解放军第二军医大学长海医院 消化内科
近年来, CCND1基因多态性与肿瘤相关性日益受到重视, 但其结果存在分歧, 可能与样本量小、种族差异和疾病遗传异质性等有关, 需要进一步扩大样本、统一研究方法及统计标准, 深入研究.
CCND1基因A870G多态是上海人群患结直肠癌的一个遗传易感分子, 可作为该地区人群结直肠癌发病风险的筛选指标.
本文选题为临床亟待解决的临床基础研究课题, 有重要临床指导意义, 但内容略显单薄.
编辑:李军亮 电编:吴鹏朕
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