CN111893189A - 检测肺癌相关基因甲基化在制备检测肺癌试剂盒中的应用 - Google Patents
检测肺癌相关基因甲基化在制备检测肺癌试剂盒中的应用 Download PDFInfo
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Abstract
本发明涉及诊断人LOC148898、VACM1、RNS4甲基化的方法和试剂盒。具体为通过协同检测三种肺癌标志物的甲基化DNA来诊断肺癌、提高肺癌检出率,所述的肺癌标志物是人LOC148898、VACM1、RNS4基因。本发明的试剂盒可以方便快捷地在分子水平上实现对肺癌的筛查,无创性、检测率高且针对性强,有利于肺癌的早期发现和及时治疗。
Description
技术领域
本发明涉及分子生物检测技术,具体地说,涉及一种与肺癌相关的多基因甲基化协同检测试剂盒。
背景技术
肺癌,又称原发性支气管癌,是对人类健康危害最大的恶性肿瘤之一,其死亡率居各类恶性肿瘤之首。在临床实践中,肺癌早期诊断一直是难点,但早期发现对癌症患者的有效治疗至关重要。目前,临床症状出现以后进行组织学和细胞学检查是肺癌诊断的金标准,但早期肺癌由于常无特殊症状而几乎不被医生和患者察觉,用常规的诊断方法也难以早期发现和早期定性诊断,并且活体取样检测也困难,严重影响了癌症的早期诊断和病人的预后。由于临床表现具有个体差异,多数患者就诊时已发展至晚期,因此早发现、早诊断、早治疗对延长肺癌恶性肿瘤患者生存期和降低死亡率有着重大意义。
与肺癌诊治有关的技术手段在不断改善,包括检测异常的基因突变,表观遗传修饰和利用免疫系统的潜力来控制肿瘤的生长等。而肿瘤标志物发展成为继影像诊断、病理诊断之后的肿瘤诊断治疗的新领域,对肿瘤的诊断、监测和治疗产生了重大影响;其中一个领域就是甲基化DNA检测。DNA甲基化是指在DNA甲基转移酶的作用下,S-腺苷甲硫氨酸的甲基(-CH3)共价结合到DNA分子的胞嘧啶(C)碱基的5位碳原子上,形成5-甲基胞嘧啶(5mC),但并不改变DNA的序列。近年来的研究表明,肺癌的发生发展与抑癌基因启动子区CPG岛甲基化导致的抑癌基因表达失活有关,且其可能发生在肺癌的早期,是一个很有潜力的早期诊断肺癌的分子指标。因此,联合检测多个基因启动子的甲基化状态,对肺癌的诊断、治疗、预后判断等方面具有重要意义。
目前现有技术中已经通过高通量测序,证明成百上千种基因的甲基化与肺癌的发生发展相关,然而高通量测序成本高昂,并且技术复杂,而目前已有的肺癌试剂盒大多为单基因检测,然而同一基因异常甲基化的DNA只占全部DNA的极小部分,大约0.1%~1%,这些非甲基化DNA与甲基化DNA只存在微小的差异,需要从高度复杂的“背景”中检测出异常的甲基化DNA。另外循环血中的DNA通常都已经降解(通常为几十到一百多碱基对)的片段,所以需要特别的抽提和检测技术才能获得较高的灵敏度。
发明内容
针对现有技术中存在的缺陷,本发明提供一种多基因甲基化协同检测试剂盒,更为具体的,所述多基因甲基化协同检测为针对人LOC148898(NCBI Reference Sequence:NM_138479.2)、VACM1(NCBI Reference Sequence:NM_003478.3)和RNS4(NCBI ReferenceSequence:NM_194430.1)基因甲基化的联合检测。
在本发明的第一方面,提供一种用于检测肺癌的试剂盒,所述的试剂盒中包括:特异性检测人LOC148898、VACM1和RNS4基因的DNA甲基化的检测试剂。作为优选的,所述试剂盒中包含检测人LOC148898、VACM1和RNS4基因甲基化的引物。更为优选的,所述LOC148898基因上游引物序列如SEQ ID NO.1所示,下游引物序列如SEQ ID NO.2所示;所述VACM1基因上游引物序列如SEQ ID NO.3所示,下游引物序列如SEQ ID NO.4所示;所述RNS4基因上游引物序列如SEQ ID NO.5所示,下游引物序列如SEQ ID NO.6所示。
在一种实施方案中,所述甲基化检测试剂盒,还包括:1×master和25mM MgCl2。
在一种实施方案中,所述甲基化检测试剂盒,还包括:阳性对照和阴性对照。
在本发明的另一方面,提供一种肺癌早期检测用甲基化标志物的检测方法,包括如下步骤:
(1)从待检组织中提取DNA,并对样本DNA、标准品DNA分别进行修饰处理,转化后的DNA(至少20ng)用水溶解后,作为PCR的模板;
(2)按照PCR体系比例加入基因的一对引物、Master、MgCl2,使得每对引物、MgCl2的终浓度分别为1uM、5mM;随后进行PCR扩增,依据不同甲基化比例的标准品DNA的解链温度差异做标准曲线,然后依据样本的PCR结果,用标准曲线得到样本的相对甲基化水平。
在一种实施方案中,所述PCR体系为:
在一种实施方案中,本发明的检测原理为:在基因的启动子区设计一对特异性的甲基化检测的引物,然后用该引物去扩增硫化样本DNA,根据PCR扩增结果的相对荧光值来确定待测样本的甲基化水平。
与常规的肺癌诊断方法相比,采用本发明的标志物进行联合检测具有快速、灵敏和特异性好等特点,可以使肺癌病人早发现、早治疗、延长生存期,具有良好的应用前景。
附图说明
附图用来提供对本发明的进一步理解,并且构成说明书的一部分,与本发明的实施例一起用于解释本发明,并不构成对本发明的限制。在附图中:
图1为三个基因以标准品DNA为模板的PCR结果绘制的标准曲线;其中,A为LOC148898、B为VACM1,C为RNS4。
具体实施方式
以下结合附图对本发明的优选实施例进行说明,应当理解,此处所描述的优选实施例仅用于说明和解释本发明,并不用于限定本发明。
实施例1肺癌相关基因甲基化标志物的特异性引物设计:
根据NCBI(美国国立生物技术信息中心)网站上公开的LOC148898、VACM1和RNS4序列信息(NCBI Reference Sequence:NM_138479.2、NCBI Reference Sequence:NM_003478.3、NCBI Reference Sequence:NM_194430.1),使用Primer Premier3.0及MethylPrimer Express v1.0设计,由上海introvigen有限公司合成。
所合成的引物信息如下:
待测基因 | 上游引物 | 下游引物 |
LOC148898 | SEQ ID NO.1 | SEQ ID NO.1 |
VACM1 | SEQ ID NO.3 | SEQ ID NO.4 |
RNS4 | SEQ ID NO.5 | SEQ ID NO.6 |
实施例2肺癌样本中LOC148898、VACM1和RNS4甲基化水平的联合检测:
实验材料:Master购自于罗氏有限公司;DNA提取试剂盒为,QIAmp DNA Mini Kit(QIAGEN);转化试剂盒为,EZ DNA Methylation kit(Zymo Research,Orange,CA,USA);试验中使用的其他材料均为常规市售种类。
实验样本:所待检测的26例肺癌组织样本均来自秦皇岛市第一医院,选用人肺癌细胞株A549、PC-9、SPCA-1和SK-MES-1作为阳性对照,正常成人染色体DNA购买自USBiological。
实验方法:
1、取大约30mg肺癌组织或正常组织剪碎后加入到5ml的离心管中,使用DNA提取试剂盒提取待测样本的DNA,然后用转化试剂盒对提取DNA进行转化;
2、PCR反应程序:
其中每组份体积:Master 10ul、25mM MgCl2 3.2ul、灭菌水5.8ul、模板1ul。
实施例3肺癌样本中其他相关基因甲基化水平的检测(对比例):
参照实施例2的PCR反应程序和检测方法,针对现有技术中已知的肺癌高度相关基因HOXA9、PCDHGB6进行检测以及分别和实施例2中的基因进行联合检测。其中所述HOXA9、PCDHGB6的引物序列分别为:
HOXA9:
上游引物:SEQ ID NO.:7
下游引物:SEQ ID NO.:8
PCDHGB6:
上游引物:SEQ ID NO.:9
下游引物:SEQ ID NO.:10
实施例4:检测结果
1.各基因在肿瘤组织和正常组织中的甲基化分布状况(附图1,p<0.01)。
2.构建ROC曲线比较4个甲基化标志物区分肺癌病人的肿瘤组织与正常组织的诊断能力。5个标志物的ROC曲线下面积分别如下:LOC148898,0.576;VACM,0.639;RNS4,0.721;PCDHGB6,0.754;HOXA9,0.583。在最佳的cutoff值下,基因的敏感性和特异性如下:LOC148898,36.5%和89.4%;VACM,52.2%和96.0%;RNS4,26.9%和88.7%;PCDHGB6,65.5%和89.1%;HOXA9,36.9%和97.3%。然而LOC148898、VACM1和RNS4三个标志物联合检测的AUC达到0.915,敏感性和特异性分别为89.9%和92.7%,而这三个标志物分别于HOXA9和PCDHGB6联用检测效果均不如LOC148898、VACM1和RNS4联用的效果。这些结果表明,LOC148898、VACM1和RNS4联用联合检测肺癌获得了意料不到的技术效果。
各基因工作特征曲线分析(ROC曲线分析)
显然,本领域的技术人员可以对本发明进行各种改动和变型而不脱离本发明的精神和范围。这样,倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内,则本发明也意图包含这些改动和变型在内。
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<110> 秦皇岛市第一医院
<120> 检测肺癌相关基因甲基化在制备检测肺癌试剂盒中的应用
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Claims (5)
1.一种用于联合检测肺癌的甲基化标志物组合,其特征在于,所述甲基化标志物为人LOC148898、VACM1和RNS4基因。
2.如权利要求1所述的甲基化标志物组合,其特征在于,所述LOC148898基因上游引物序列如SEQ ID NO.1所示,下游引物序列如SEQ ID NO.2所示;所述VACM1基因上游引物序列如SEQ ID NO.3所示,下游引物序列如SEQ ID NO.4所示;所述RNS4基因上游引物序列如SEQID NO.5所示,下游引物序列如SEQ ID NO.6所示。
3.一种用于检测肺癌的甲基化试剂盒,其特征在于,所述试剂盒中包含SEQ ID NO.:1-6所示的引物序列组合。
4.如权利要求3所述的用于检测肺癌的甲基化试剂盒,其特征在于,所述试剂盒中还包括阳性对照和阴性对照。
5.如权利要求4所述的用于检测肺癌的甲基化试剂盒,其特征在于,所述试剂盒中还包括:1×master和25mM MgCl2。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090123925A1 (en) * | 2005-09-23 | 2009-05-14 | Collie-Duguid Elaina S R | Cancer therapy prognosis and target |
US20100184052A1 (en) * | 2007-06-01 | 2010-07-22 | Paul Roepman | Prognostic gene expression signature for non small cell lung cancer patients |
US20160153053A1 (en) * | 2010-08-31 | 2016-06-02 | The General Hospital Corporation | Cancer-related biological materials in microvesicles |
CN108548923A (zh) * | 2018-04-20 | 2018-09-18 | 山东省千佛山医院 | 肺小细胞癌早期特异性自身抗体panel诊断试剂盒 |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090123925A1 (en) * | 2005-09-23 | 2009-05-14 | Collie-Duguid Elaina S R | Cancer therapy prognosis and target |
US20100184052A1 (en) * | 2007-06-01 | 2010-07-22 | Paul Roepman | Prognostic gene expression signature for non small cell lung cancer patients |
US20160153053A1 (en) * | 2010-08-31 | 2016-06-02 | The General Hospital Corporation | Cancer-related biological materials in microvesicles |
CN108548923A (zh) * | 2018-04-20 | 2018-09-18 | 山东省千佛山医院 | 肺小细胞癌早期特异性自身抗体panel诊断试剂盒 |
Non-Patent Citations (2)
Title |
---|
FENG GAO, ET AL.: ""The functions and properties of cullin-5, a potential therapeutic target for cancers"", 《AM J TRANSL RES》 * |
张艳秋: ""肺癌相关非编码RNA的筛选及功能研究"", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
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