MicroRNA4293 基因多态性与女性肝癌易感性相关

蔡敏　郑万威　张骏　程烽涛　李力



·论著·

MicroRNA4293 基因多态性与女性肝癌易感性相关

蔡敏郑万威张骏程烽涛李力

200090上海同济大学附属杨浦医院消化科(蔡敏，程烽涛，李力)；复旦大学附属华山医院消化科(郑万威，张骏)

【摘要】目的研究microRNA-4293的SNP(rs12220909)与原发性肝癌易感性的关系。方法收集人体血液样本1788份，样本取自健康对照者884名，原发性肝癌患者904例。抽提血液样本中基因组DNA，应用MassARRAY法对microRNA-4293的SNP(rs12220909)位点进行基因分型；应用二元逻辑回归法分析microRNA-4293的SNP(rs12220909)位点与肝癌易感性之间的关系。结果女性肝癌患者microRNA-4293的SNP(rs12220909)中CC基因型较健康对照组显著增加(P=0.04)，在HBV相关肝癌中显著性进一步增加(P=0.02)，提示CC基因可能增加女性肝癌尤其是HBV相关肝癌的易感性。而在男性患者中，microRNA-4293的SNP(rs12220909)与肝癌易感性并不相关(P=0.33)。结论microRNA-4293的SNP(rs12220909)与女性肝癌易感性相关。

【关键词】原发性肝癌；易感性；MicroRNA-4923；SNP(rs12220909)

原发性肝癌是最常见的肝脏恶性肿瘤，具有发病率高、病死率高、易复发、易转移的特点，已严重威胁人类的健康[1,2]。由于大部分肝癌患者确诊时病情已进展至晚期，预后较差。尽管肝癌有多种治疗方法，如手术切除、介入治疗、肝移植等，但因肝功能异常和肿瘤过大等因素，只有5%～15%的患者适合手术切除治疗[3]。

microRNA(miRNA)是一类由22 个核苷酸组成的非编码单链RNA 分子，参与转录后基因表达调控。miRNA通过对靶标mRNA转录后去稳定和(或)抑制其翻译，来调节细胞增殖、分化、迁移和凋亡等功能[4]。研究发现，多种与肿瘤相关的miRNA，其靶标基因与肿瘤的发生、发展相关[5]。单核苷酸多态性(single nucleotide polymorphisms，SNP)已成为研究复杂遗传性疾病、药物基因组学和人类进化的第三代遗传标记。因为肿瘤相关基因的SNP常影响该基因的表达及功能[6]，能够改变肿瘤易感性，故此类基因SNP常作为生物学标志物应用于肿瘤研究[7]。针对肝癌miRNA表达谱的研究发现，miRNA的微小变化可引起靶mRNA翻译的显著变化，从而影响到肝癌的发生和进展[8]。

mir-4293位于第10号染色体上，目前研究发现mir-4293的SNP(rs12220909)与食管鳞状细胞癌易感性相关[9]，但关于该miRNA的研究较少，其与肝癌的相关性仍未知。

本研究对血液样本进行mir-4293的SNP(rs12220909)检测，并分析该位点与原发性肝癌易感性的关系。

资料和方法

一、样本收集

共收集人体血液样本1788份。样本取自健康者884名，原发性肝癌患者904例，其中HBV相关肝癌701例。健康人群取自江苏泰州地区，无肿瘤及肝炎病史；原发性肝癌病例取自复旦大学附属华山医院、同济大学附属杨浦医院及东方肝胆医院，所有肝癌患者均经病理确诊。

二、核酸提取和基因分型

应用血液基因组DNA抽提试剂盒(Axygen Biosciences, Union City, USA)从血液样本中抽提基因组DNA。在测序分析前，应用NanoDrop 2000c分光光度计测定DNA浓度，并用电泳法测定其纯度。应用MassARRAY法对mir-4293的SNP(rs12220909)位点进行基因分型。mir-4293的SNP(rs12220909)的染色体起止位点为chr10：14425199-14425276，SNP位置为14425221(mat)，扩增引物为ACGTTGGATGGCTGGTGACATTGCTAATTCA-CGTTGGATGGGAGAGGGGAAATCCTATTT，延伸引物为：CCCAAGGCTGTTCCTGTCA。

三、数据分析

应用二元逻辑回归法分析mir-4293的SNP(rs12220909)位点与肝癌易感性之间的关系。P<0.05为差异有统计学意义。所有统计学分析采用SPSS 16.0软件。

结果

健康对照组884名(男性644例，女性240例)，原发性肝癌患者904例(男性742例，女性162例)。研究对象的基本信息见表1。应用二元逻辑回归法，校正年龄、吸烟和饮酒等其他因素影响后发现，mir-4293的SNP(rs12220909)位点的各基因型在肝癌组与对照组差异无统计学意义(P=0.07)，见表2。在女性患者中，肝癌组CC基因型较对照组显著增加(P=0.04)，在HBV相关肝癌中显著性进一步增加(P=0.02)，见表3；而在男性患者中各基因型在肝癌组与对照组差异无统计学意义，见表4。

表1　研究对象的基本信息

表2　Mir-4293的SNP(rs12220909)与肝癌易感性分析

表3　女性患者中Mir-4293的SNP(rs12220909)与肝癌易感性分析

表4　男性患者中Mir-4293的SNP(rs12220909)与肝癌易感性分析

讨论

早期诊断和有效预防肝癌的已成为世界性的医疗问题，在目前的临床应用中迫切需要开发新型有效的标志物用于肝癌的筛查、早期诊断和判断预后，并需积极寻找新的治疗靶标与方法[10]。研究发现，miRNA表达紊乱在多种肿瘤，包括原发性肝癌的发生、发展中起到重要作用[11]。而且，一些miRNA在肝癌中特异性表达，并且与多种肿瘤基因如P53、PTENZ和RA等组成了调控网络，参与肝癌的发生、侵袭和转移[12-14]。miRNA的SNP能够影响miRNA的成熟及miRNA与靶标基因的结合，从而影响靶标基因的功能，进一步影响肝癌、非小细胞肺癌等恶性肿瘤的易感性、治疗反应及预后[15,16]。综上，miRNA及其SNP位点具有极大的潜力成为肝癌筛查、早期诊断及预后判断的新型标志物，同时为肝癌预防及治疗的研究提供新的靶标。故对于miRNA的SNP与肝癌的研究就显得尤为重要[17-19]。

本研究发现位于第10号染色体上mir-4293的SNP(rs12220909)与女性肝癌易感性相关，女性肝癌患者中CC基因型较健康对照组显著增加(P=0.04)，在HBV相关肝癌中显著性进一步增加(P=0.02)，提示CC基因可能增加女性肝癌尤其是HBV相关肝癌的易感性。而在男性患者中，mir-4293的SNP(rs12220909)与肝癌易感性并不相关。

综上所述，mir-4293的SNP(rs12220909)中CC基因型能够显著增加女性原发性肝癌及HBV相关肝癌的易感性，而在男性患者中并无显著作用。本研究可为肝癌发病性别差异的机制研究提供新方向，同时有助于推动miRNA SNP在肝癌早期诊断上的临床应用。

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(本文编辑：赖荣陶)

通信作者：李力，Email: lli10437@sina.com

Corresponding author:LI Li, Email: lli10437@sina.com

(收稿日期：2016-03-31)

Genetic polymorphisms of microRNA-4293 and hepatocellular carcinoma susceptibility in female

CAIMin,ZHENGWan-wei,ZHANGJun,CHENGFeng-tao,LILi.

TongjiUniversitySchoolofMedicine,YangpuHospital,DepartmentofDigestiveDiseases,Shanghai200090,China

【Abstract】ObjectiveTo investigate the association between single nucleotide polymorphism (SNP) mapping to microRNA-4293 (rs12220909) and hepatocellular carcinoma (HCC) susceptibility. MethodsBlood samples were collected from 1788 cases, including 884 normal control cases and 904 HCC cases. Genomic deoxyribonucleic acid was extracted from blood samples, and SNPs mapping to microRNA-4293 (rs12220909) were detected by MassARRAY. Then association between genotype of rs12220909 and HCC susceptibility was analyzed by binary logistic regression. ResultsIn female HCC patients, CC genotype of rs12220909, especially in hepatitis B virus (HBV)-related HCC patients (P=0.02), was significantly higher than that in healthy control group (P=0.04). It indicated that CC genotype might be a risk factor for female HCC, especially for HBV-related HCC. However, this association did not exist in male patients (P=0.33). ConclusionGenotype of microRNA-4293 (rs12220909) is relevant to HCC in female patients.

【Key words】Hepatocellular carcinoma; Susceptibility; MicroRNA-4923; SNP(rs12220909)