CN107012213A - 结直肠癌的生物标记物 - Google Patents
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Abstract
结直肠癌的生物标记物。涉及干扰素诱导的跨膜蛋白1在抑制结直肠癌中内源性逆转录病毒中的应用。用CRISPR/Cas9技术构建IFITM1‑KO的hESCs来研究IFITM1的功能。研究发现,IFITM1缺失的人胚胎干细胞(hESCs)在细胞增殖,细胞多能性,端粒长度,端粒酶活性等方面与IFITM1‑WT相差不多。IFITM1‑KO hESCs中人类内源性逆转录病毒表达升高,并且其在癌旁组织中表达比结直肠癌组织表达升高。通过ChIP‑qPCR检测,发现在IFITM1‑KO hESCs中H3K9me3在HERVs位点富集下降。这些数据表明在hESCs和结直肠癌中,IFITM1表达水平与HERVs表达呈负相关,IFITM1通过调控表观遗传来抑制HERVs的表达。
Description
技术领域:
本发明属于生物医学技术领域,具体涉及基于检测IFITM1及其调控的HERVs相关基因的表达水平来诊断结直肠癌的方法。
背景技术:
1.近年来,多项研究表明IFITM1在多种恶性肿瘤中高表达,如IFITM1在子***、食管癌、卵巢癌、脑癌中均高表达,并且在结直肠癌中表达上调并被认为是结直肠癌分子标志物。越来越多人关注其在恶性肿瘤的发生发展及肿瘤早期侵袭中的作用。在结直肠癌中,其仅在肿瘤组织有IFITM1表达,在肠道肿瘤形成过程中Wnt/β-catenin信号通路的激活能诱导IFITM1的表达;在慢性髓系白血病中,其在高危组表达水平低,低危组表达水平高,表达水平高的患者具有更好的预后,因此,有人认为IFITM1可能是慢性髓系白血病患者预后的分子标志物。
2.人内源性逆转录酶病毒(Human endogenous retroviruses,HERVs)是可转座的遗传因子,可以***人的基因组中,在人基因组中占到8%的比例。其在胚胎发育过程中被激活,IFITM1能保护胚胎和生殖细胞免受内源性逆转录酶病毒和外源性病毒的感染。ERVs包含有长的末端重复序列(LTRs),能通过H3K9me3组蛋白修饰来抑制其表达,并且可以通过增加基因组DNA甲基化来抑制HERVs的表达。HERVs过表达会导致基因组不稳定,异常表达的HERVs甚至与癌症的发生相关联。
3.前人研究中提议将IFITM1作为结直肠癌分子标志物,经过试验发现在结直肠癌中IFITM1的表达与HERVs表达呈负相关,本发明中以IFITM1及HERVs的表达作为双重标准来作为结直肠癌的标志物。
发明内容:
本发明的目的在于提供一种IFITM1及其调控的HERVs在结直肠癌标记中的用途。具体说,本发明涉及IFITM1蛋白及其调控的HERVs在结直肠癌诊断试剂盒中的应用。本发明的实施能提供更加准确的结直肠癌的诊断。
本发明的技术方案
一种蛋白及其调控的基因即人内源性逆转录病毒在制备用于检测受试者中存在结直肠癌的体外方法中的试剂盒中的用途,该用途包括:检测该IFITM1及其调控的基因即人内源性逆转录病毒(HERVs)在受试者的测定细胞即结直肠活检测样品中的表达;将所述的测定值与参考值相比较,IFITM1明显高于参考值并且HERVs明显低于参考值,表明受试者可能患有结直肠癌。
所述参考值水平是IFITM1和HERVs相关基因在正常细胞即结直肠活检测样品中的水平。所述正常细胞是相同受试者中与测定细胞相同类型的细胞。所述正常细胞是受试者中与测定细胞相同类型的细胞,所述受试者没有癌症。所述样品已知或被怀疑包含肿瘤细胞。
1.本发明至少部分基于对在人结直肠中高表达的IFITM1及在结直肠癌肿瘤中普遍低表达的HERVs基因的鉴定。本发明的方法对于结直肠癌的检测有用。
2.利用IFITM1和HERVs作为生物标记诊断结直肠癌。
3.本申请所描述的方法可用于诊断受试者中的结直肠癌的存在。在一些具体实施方案中,所述的方法包括,从受试者获取样品,评估所述样品中IFITM1和HERVs存在和/或水平,将所述的存在和/或水平与一个或以上的参照相比较,例如代表正常IFITM1和HERVs水平的对照参照,如来自未受影响的受试者或来自相同个体的正常细胞的中的水平。
4.样品:在本发明的方法的一些具体实施方案中,所述的样品是包括已知的或被怀疑的肿瘤细胞,例如是活检样品。在一些具体实施方案中,所述的样品是冷冻的,固定的和/或经渗透处理的,例如是***固定石蜡包埋(FFPE)的样品或冰冻在液氮中的样品。
5.检测的方法:a.可以使用本领域任何已知的方法来检测和/或定量本申请所述的生物标记的水平。例如可以利用本领域已知的方法评估IFITM1和HERVs的mRNA(转录物)的水平,例如Northern印迹,RNA原位杂交(RNA-ISH),RNA表达测定,例如微阵列分析,RT-PCR,RNA测序(例如利用随机引物或寡T引物),深度测序(deep sequencing),克隆,Northern印迹,和扩增转录物,例如利用定量实时聚合酶链式反应(qRT-PCR)。b.在一些具体实施方案中,可以检测IFITM1和HERVs编码的蛋白质的水平。可利用本领域已知的方法评估蛋白质的存在和/或水平,例如利用定量免疫分析方法,如酶联免疫吸附试验(ELISAs),免疫沉淀,免疫荧光,免疫组织化学,酶免疫测定(EIA),放射免疫测定(RIA)和Western印迹分析。c.在一些具体实施方案中,所述的方法包括使选择性地与生物标记结合的试剂与样品接触,以评估样品中所述生物标记,例如IFITM1及HERVs转录物/mRNA或蛋白质(例如寡核苷酸探针,抗体或其抗原结合部分)的水平。在一些具体实施方案中,所述的试剂带有可检测的标记。关于经标记的试剂,包括将所述试剂与可检测的物质相偶联(即物理连接)的直接标记,以及通过所述试剂与可检测的物质的反应性进行间接标记。可检测的物质的示例是本领域已知的,包括化学发光的,荧光的,放射性的或显色的标记。例如可检测的物质包括各种酶,辅基,荧光材料,发光材料,生物发光材料和放射性材料。合适的酶的例子包括辣根过氧化物酶,碱性磷酸酶,β-半乳糖苷酶,或乙酰胆碱酯酶;合适的辅基复合物的例子包括链霉抗生物素蛋白/生物素,以及亲和素/生物素;合适的荧光物质的例子包括伞形酮,荧光素,异硫氰酸荧光素,罗丹明),二氯三嗪氨荧光素(dichlorotriazinylaminefluorescein),丹磺酰氯,量子点(quantum dots)或藻红蛋白;发光材料的示例包括鲁米诺;生物发光材料的示例包括荧光素酶,荧光素和水母发光蛋白;
合适的放射性物质的示例包括125I,131I,35S or3H。总体而言,可利用抗体检测蛋白质。抗体可以是多克隆抗体,更优选单克隆抗体。可应用完整抗体或其抗原结合片段(例如Fab或F(ab')2)。
附图说明:
图1是免疫荧光图和柱状图,通过免疫荧光和定量PCR实验证明IFITM1在结直肠癌肿瘤部位和hESCs(WA26和RUES2两个细胞系)中确实高表达。
图2是包括免疫荧光图(D,E),柱状图(G,I),凝胶电泳图(H),细胞形态图(C),曝光图片(B,F,J)。图2验证构建的IFITM1敲除细胞系,并检验IFITM1敲除后细胞变化。其中A显示CRISPR/Cas9敲除基因原理图。B,D鉴定IFITM1敲除细胞系。C显示IFITM1敲除后细胞形态无变化。F显示IFITM1敲除后干细胞多能性无明显变化。I,J显示IFITM1敲除后细胞端粒无明显变化。G,H显示IFITM1敲除后细胞端粒酶活性物明显变化。
图3是柱状图,检测IFITM1敲除后细胞内HERVs的表达。其中A显示HERVs的结构,黑色箭头表明我们所设计定量PCR(ChIP-qPCR引物同定量PCR)引物的大致位点。B显示在hESCs中,IFITM1敲除后HERVs表达升高(P15代次)。C通过ChIP-qPCR实验说明在hESCs(RUES2)中,IFITM1敲除后H3K9me3在HERVs位点的结合下降(P15代次)。
图4是免疫荧光图和散点图,检测IFITM1敲除后细胞内DNA甲基化变化情况。通过免疫荧光实验,并且统计细胞荧光强度后可以知道5mC/5hmC在IFIFM1-KO的hESCs中下降(即DNA甲基化下降),则可以说明IFITM1通过调控DNA甲基化来调控HERVs的表达。
图5是柱状图,检测病人样品中HERVs的表达。说明在结直肠癌中肿瘤部位HERVs的表达比正常部位低。
图6是免疫荧光图和散点图,检测肿瘤样品中DNA甲基化情况。说明在结直肠癌中,通过5mC/5hmC的比值可以知道肿瘤部位5mC(DNA甲基化程度高)升高,DNA甲基化能抑制HERVs的表达,所以IFITM1能通过调控DNA甲基化程度来抑制HERVs的表达。
具体实施例:
下面结合具体实施例,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用限制本发明的范围。除非另有描述,本发明的实施将采用分子生物学、和免疫学的常规技术,这些均是本领域技术人员所知的。
实施例1:
检测IFITM1在结直肠癌中和hESCs中的表达(图1)
材料与方法:
1.随机选取结直肠癌病人,取结直肠癌病人冰冻组织切片(包括肿瘤部位和正常部位),进行免疫荧光染色。
2.随机选取结直肠癌病人,提取RNA,通过定量PCR检测其IFITM1的mRNA水平;并取hESCs细胞系(WA26和RUES2)RNA,检测IFITM1表达水平。
3.结果:IFITM1在结直肠癌中肿瘤部位表达量显著高于正常组织,并且其在hESCs细胞系中表达高(以HEF细胞作为对照)。
实施例2:
利用CRISPR/Cas9技术构建IFITM1敲除的hESCs细胞系(图2)
1.细胞培养:利用Essential 8(invitrogen)培养液在37℃、5%CO2条件下培养培养hESCs细胞,需要每天换液。
2.构建质粒:根据NCBI公布的IFITM1蛋白的CDS区序列设计引物,通过扩增、连接等将其***px459质粒中,经阳性克隆PCR鉴定、测序、BLAST比对,结果显示px459-IFITM1构建成功。
3.细胞转染及鉴定:hESCs细胞系在Essential 8培养液中,于37℃、5%CO2培养箱中培养。利用核转技术将质粒转到细胞中,然后培养细胞。通过药物筛选及挑取单克隆细胞,然后经过测序及western blot实验和免疫荧光实验验证得到IFITM1敲除细胞。
实施例3:
验证IFITM1敲除后对hESCs的影响(图2,3,4)
1.通过western blot实验检测到IFITM1敲除后细胞多能性没有变化;通过定量PCR实验和TRAP实验检测到IFITM1敲除后细胞端粒酶没有变化;通过定量PCR(T/S ratio)实验检测到细胞端粒长度没有变化。
2.通过PCR实验我们检测到IFITM1敲除后,收取细胞提取RNA,检测知道细胞内HERVs升高。检测用到的引物如下:
3.通过ChIP-qPCR实验(用2中引物),我们发现在IFITM1敲除后,H3K9me3在HERVs位点富集程度下降,因为H3K9me3具有抑制HERVs表达的作用,因此导致HERVs表达升高。
4.收集3中同代次细胞做免疫荧光(5mC和5hmC),统计细胞的荧光强度,通过5mC/5hmC比值可以知道,IFITM1敲除后细胞DNA甲基化程度降低,DNA甲基化能够抑制抑制HERVs的表达。
5结论:IFITM1能通过调节表观遗传的方式来抑制HERVs的表达。
实施例4:
验证结直肠中HERVs的表达(图5,6)
1.取结直肠癌病人肿瘤组织和正常组织,然后提取RNA,通过定量PCR检测发现,在肿瘤组织中(IFITM1表达高)HERVs表达比正常组织低(IFITM1表达低)。
2.将组织石蜡包埋后切片后,免疫荧光染色(5mC和5hmC),统计细胞的荧光强度,通过5mC/5hmC比值可以知道,肿瘤组织中(IFITM1表达高)DNA甲基化程度高。
3.结论:同hESCs中一样,IFITM1能通过调节表观遗传的方式来抑制HERVs的表达,并且确定结直肠中肿瘤组织IFITM1表达高,并且HERVs表达降低。
Claims (5)
1.一种结直肠癌的生物标记物,涉及蛋白及其调控的基因即人内源性逆转录病毒在制备用于检测受试者中存在结直肠癌的体外方法中的试剂盒中的用途,该用途包括:检测该IFITM1及其调控的基因即人内源性逆转录病毒(HERVs)在受试者的测定细胞即结直肠活检测样品中的表达;将所述的测定值与参考值相比较,IFITM1明显高于参考值并且HERVs明显低于参考值,表明受试者可能患有结直肠癌。
2.根据权利要求1所述的用途,其中所述参考值水平是IFITM1和HERVs相关基因在正常细胞即结直肠活检测样品中的水平。
3.根据权利要求2所述的用途,其中所述正常细胞是相同受试者中与测定细胞相同类型的细胞。
4.根据权利要求2所述的用途,其中所述正常细胞是受试者中与测定细胞相同类型的细胞,所述受试者没有癌症。
5.根据权利要求1所述的用途,其中所述样品已知或被怀疑包含肿瘤细胞。
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Cited By (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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US11542496B2 (en) | 2017-03-10 | 2023-01-03 | President And Fellows Of Harvard College | Cytosine to guanine base editor |
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US11912985B2 (en) | 2020-05-08 | 2024-02-27 | The Broad Institute, Inc. | Methods and compositions for simultaneous editing of both strands of a target double-stranded nucleotide sequence |
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102215870A (zh) * | 2008-09-18 | 2011-10-12 | 学校法人庆应义塾 | 癌症的诊断方法和治疗方法 |
US20140024547A1 (en) * | 2006-11-13 | 2014-01-23 | Genenews Corporation | Gene Expression Profiling For Identification, Monitoring And Treatment Of Colorectal Cancer |
CN104185685A (zh) * | 2011-12-20 | 2014-12-03 | 拜奥默里克斯公司 | 用于结肠癌的体外诊断或预后的方法 |
-
2017
- 2017-03-24 CN CN201710185973.0A patent/CN107012213A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140024547A1 (en) * | 2006-11-13 | 2014-01-23 | Genenews Corporation | Gene Expression Profiling For Identification, Monitoring And Treatment Of Colorectal Cancer |
CN102215870A (zh) * | 2008-09-18 | 2011-10-12 | 学校法人庆应义塾 | 癌症的诊断方法和治疗方法 |
CN104185685A (zh) * | 2011-12-20 | 2014-12-03 | 拜奥默里克斯公司 | 用于结肠癌的体外诊断或预后的方法 |
Non-Patent Citations (5)
Title |
---|
AUDREY T LIN等: "Role of Endogenous Retroviruses in Human Genetic Diseases", 《GENETICS & DISEASE》 * |
EDWARD J.GROW等: "Intrinsic retroviral reactivation in human preimplantation embryos and pluripotent cells", 《NATURE》 * |
PAULINE ANDREU等: "Identification of the IFITM Family as a New Molecular Marker in Human Colorectal Tumors", 《CANCER RESEARCH》 * |
于红莲: "HERV参与肿瘤发生的研究进展", 《济宁医学院学报》 * |
于跃明等: "《结直肠癌》", 28 February 2010, 科学技术文献出版社 * |
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