MicroRNA4293 基因多態(tài)性與女性肝癌易感性相關(guān)

蔡敏　鄭萬威　張駿　程烽濤　李力

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·論著·

MicroRNA4293 基因多態(tài)性與女性肝癌易感性相關(guān)

蔡敏鄭萬威張駿程烽濤李力

200090上海同濟大學(xué)附屬楊浦醫(yī)院消化科(蔡敏，程烽濤，李力)；復(fù)旦大學(xué)附屬華山醫(yī)院消化科(鄭萬威，張駿)

【摘要】目的研究microRNA-4293的SNP(rs12220909)與原發(fā)性肝癌易感性的關(guān)系。方法收集人體血液樣本1788份，樣本取自健康對照者884名，原發(fā)性肝癌患者904例。抽提血液樣本中基因組DNA，應(yīng)用MassARRAY法對microRNA-4293的SNP(rs12220909)位點進行基因分型；應(yīng)用二元邏輯回歸法分析microRNA-4293的SNP(rs12220909)位點與肝癌易感性之間的關(guān)系。結(jié)果女性肝癌患者microRNA-4293的SNP(rs12220909)中CC基因型較健康對照組顯著增加(P=0.04)，在HBV相關(guān)肝癌中顯著性進一步增加(P=0.02)，提示CC基因可能增加女性肝癌尤其是HBV相關(guān)肝癌的易感性。而在男性患者中，microRNA-4293的SNP(rs12220909)與肝癌易感性并不相關(guān)(P=0.33)。結(jié)論microRNA-4293的SNP(rs12220909)與女性肝癌易感性相關(guān)。

【關(guān)鍵詞】原發(fā)性肝癌；易感性；MicroRNA-4923；SNP(rs12220909)

原發(fā)性肝癌是最常見的肝臟惡性腫瘤，具有發(fā)病率高、病死率高、易復(fù)發(fā)、易轉(zhuǎn)移的特點，已嚴(yán)重威脅人類的健康[1,2]。由于大部分肝癌患者確診時病情已進展至晚期，預(yù)后較差。盡管肝癌有多種治療方法，如手術(shù)切除、介入治療、肝移植等，但因肝功能異常和腫瘤過大等因素，只有5%～15%的患者適合手術(shù)切除治療[3]。

microRNA(miRNA)是一類由22 個核苷酸組成的非編碼單鏈RNA 分子，參與轉(zhuǎn)錄后基因表達(dá)調(diào)控。miRNA通過對靶標(biāo)mRNA轉(zhuǎn)錄后去穩(wěn)定和(或)抑制其翻譯，來調(diào)節(jié)細(xì)胞增殖、分化、遷移和凋亡等功能[4]。研究發(fā)現(xiàn)，多種與腫瘤相關(guān)的miRNA，其靶標(biāo)基因與腫瘤的發(fā)生、發(fā)展相關(guān)[5]。單核苷酸多態(tài)性(single nucleotide polymorphisms，SNP)已成為研究復(fù)雜遺傳性疾病、藥物基因組學(xué)和人類進化的第三代遺傳標(biāo)記。因為腫瘤相關(guān)基因的SNP常影響該基因的表達(dá)及功能[6]，能夠改變腫瘤易感性，故此類基因SNP常作為生物學(xué)標(biāo)志物應(yīng)用于腫瘤研究[7]。針對肝癌miRNA表達(dá)譜的研究發(fā)現(xiàn)，miRNA的微小變化可引起靶mRNA翻譯的顯著變化，從而影響到肝癌的發(fā)生和進展[8]。

mir-4293位于第10號染色體上，目前研究發(fā)現(xiàn)mir-4293的SNP(rs12220909)與食管鱗狀細(xì)胞癌易感性相關(guān)[9]，但關(guān)于該miRNA的研究較少，其與肝癌的相關(guān)性仍未知。

本研究對血液樣本進行mir-4293的SNP(rs12220909)檢測，并分析該位點與原發(fā)性肝癌易感性的關(guān)系。

資料和方法

一、樣本收集

共收集人體血液樣本1788份。樣本取自健康者884名，原發(fā)性肝癌患者904例，其中HBV相關(guān)肝癌701例。健康人群取自江蘇泰州地區(qū)，無腫瘤及肝炎病史；原發(fā)性肝癌病例取自復(fù)旦大學(xué)附屬華山醫(yī)院、同濟大學(xué)附屬楊浦醫(yī)院及東方肝膽醫(yī)院，所有肝癌患者均經(jīng)病理確診。

二、核酸提取和基因分型

應(yīng)用血液基因組DNA抽提試劑盒(Axygen Biosciences, Union City, USA)從血液樣本中抽提基因組DNA。在測序分析前，應(yīng)用NanoDrop 2000c分光光度計測定DNA濃度，并用電泳法測定其純度。應(yīng)用MassARRAY法對mir-4293的SNP(rs12220909)位點進行基因分型。mir-4293的SNP(rs12220909)的染色體起止位點為chr10：14425199-14425276，SNP位置為14425221(mat)，擴增引物為ACGTTGGATGGCTGGTGACATTGCTAATTCA-CGTTGGATGGGAGAGGGGAAATCCTATTT，延伸引物為：CCCAAGGCTGTTCCTGTCA。

三、數(shù)據(jù)分析

應(yīng)用二元邏輯回歸法分析mir-4293的SNP(rs12220909)位點與肝癌易感性之間的關(guān)系。P<0.05為差異有統(tǒng)計學(xué)意義。所有統(tǒng)計學(xué)分析采用SPSS 16.0軟件。

結(jié)果

健康對照組884名(男性644例，女性240例)，原發(fā)性肝癌患者904例(男性742例，女性162例)。研究對象的基本信息見表1。應(yīng)用二元邏輯回歸法，校正年齡、吸煙和飲酒等其他因素影響后發(fā)現(xiàn)，mir-4293的SNP(rs12220909)位點的各基因型在肝癌組與對照組差異無統(tǒng)計學(xué)意義(P=0.07)，見表2。在女性患者中，肝癌組CC基因型較對照組顯著增加(P=0.04)，在HBV相關(guān)肝癌中顯著性進一步增加(P=0.02)，見表3；而在男性患者中各基因型在肝癌組與對照組差異無統(tǒng)計學(xué)意義，見表4。

表1　研究對象的基本信息

表2　Mir-4293的SNP(rs12220909)與肝癌易感性分析

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

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

討論

早期診斷和有效預(yù)防肝癌的已成為世界性的醫(yī)療問題，在目前的臨床應(yīng)用中迫切需要開發(fā)新型有效的標(biāo)志物用于肝癌的篩查、早期診斷和判斷預(yù)后，并需積極尋找新的治療靶標(biāo)與方法[10]。研究發(fā)現(xiàn)，miRNA表達(dá)紊亂在多種腫瘤，包括原發(fā)性肝癌的發(fā)生、發(fā)展中起到重要作用[11]。而且，一些miRNA在肝癌中特異性表達(dá)，并且與多種腫瘤基因如P53、PTENZ和RA等組成了調(diào)控網(wǎng)絡(luò)，參與肝癌的發(fā)生、侵襲和轉(zhuǎn)移[12-14]。miRNA的SNP能夠影響miRNA的成熟及miRNA與靶標(biāo)基因的結(jié)合，從而影響靶標(biāo)基因的功能，進一步影響肝癌、非小細(xì)胞肺癌等惡性腫瘤的易感性、治療反應(yīng)及預(yù)后[15,16]。綜上，miRNA及其SNP位點具有極大的潛力成為肝癌篩查、早期診斷及預(yù)后判斷的新型標(biāo)志物，同時為肝癌預(yù)防及治療的研究提供新的靶標(biāo)。故對于miRNA的SNP與肝癌的研究就顯得尤為重要[17-19]。

本研究發(fā)現(xiàn)位于第10號染色體上mir-4293的SNP(rs12220909)與女性肝癌易感性相關(guān)，女性肝癌患者中CC基因型較健康對照組顯著增加(P=0.04)，在HBV相關(guān)肝癌中顯著性進一步增加(P=0.02)，提示CC基因可能增加女性肝癌尤其是HBV相關(guān)肝癌的易感性。而在男性患者中，mir-4293的SNP(rs12220909)與肝癌易感性并不相關(guān)。

綜上所述，mir-4293的SNP(rs12220909)中CC基因型能夠顯著增加女性原發(fā)性肝癌及HBV相關(guān)肝癌的易感性，而在男性患者中并無顯著作用。本研究可為肝癌發(fā)病性別差異的機制研究提供新方向，同時有助于推動miRNA SNP在肝癌早期診斷上的臨床應(yīng)用。

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(本文編輯：賴榮陶)

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)