CN107868788A - 一组多梳蛋白靶基因及其筛选方法和应用 - Google Patents

一组多梳蛋白靶基因及其筛选方法和应用 Download PDF

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CN107868788A
CN107868788A CN201711127622.0A CN201711127622A CN107868788A CN 107868788 A CN107868788 A CN 107868788A CN 201711127622 A CN201711127622 A CN 201711127622A CN 107868788 A CN107868788 A CN 107868788A
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谢浩
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廖艳林
王明滢
陈萌萌
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Abstract

本发明公开了一组多梳蛋白靶基因及其筛选方法和应用。首先在ModENCODE数据库中检索果蝇胚胎PcG结合相关的ChIP‑seq数据，找到有PcG结合的靶基因。结合PcGRNAi之后的RNA‑seq的数据以及统计分析，筛选得到PcG特异结合的相关的基因。本发明通过搜集已有的ChIP‑seq数据，结合PcG与靶基因之间的调控关系以及芯片等数据，筛选得到的基因可以作为PcG直接结合并发挥调控功能的基因，这些基因在细胞的增殖分化中起重要作用，同时可以用来筛选潜在的原癌基因。

Description

一组多梳蛋白靶基因及其筛选方法和应用
技术领域
本发明属于生物技术领域，具体涉及一组多梳蛋白靶基因及其筛选方法和应用。
背景技术
近些年来关于癌基因的研究是一大热点，发现了很多癌症相关的基因。截止目前，常见的癌基因家族有：一、src家族，它们都含有相似的基因编码结构，产物具有使酪氨酸磷酸化的蛋白激酶活性，定位于胞膜内面或跨膜分布；二、ras家族，家族成员基因序列差异大，但所编码的蛋白质是P21，位于细胞质膜内面，P21可与GTP结合，有GTP酶活性，并参与cAMP水平的调节；三、myc家族，编码核内DNA结合蛋白，有直接调节其他基因转录的作用；四、sis家族，只有sis基因一个成员，编码P28，与人血小板源生长因子结构类似，刺激间叶组织细胞分裂增殖；五、myb家族，编码核蛋白，能与DNA结合，为核内的一种转录因子。
据报道，多梳蛋白(Polycomb group proteins，PcG)与乳腺癌和肺癌有关。PcG靶基因在细胞分化、增殖、凋亡、生物生长发育及人类疾病发生、发展等过程中发挥着不可忽视的作用。
发明内容
解决的技术问题：本发明的目的是提供一组多梳蛋白靶基因及其筛选方法和应用，筛选得到的靶基因可以作为PcG直接结合并发挥调控功能的基因，这些基因在细胞的增殖分化中起重要作用，同时可以用来筛选潜在的原癌基因。
技术方案：
一组多梳蛋白靶基因，包括ococ、nub、iab-4、btd、opa、grn、retn、Dll、unpg、mab-21、Kr、disco-r、so、E5、ap、drm、dac、ems、slp2、Vsx1、Ptx1、Oaz、Lim1、Vsx2、dve、rpr、pdm2、lab、pros、wg、peb、Sox100B、tsh、kn、CG14762、Drl-2、CG31778、CG8157、Psc、Sox102F、CG13321、ss、Su(z)2、KP78a、GATAe、klu、KP78b、gsb-n、D、slgA、elB、lz、zfh2、Apt、bab2、CG32121、SoxN、CG10357、Hexo2、mirr、Ama、Zasp52、modSP、fkh、tio、CG33465、CG5613、CG9328、Sr-CIV、Antp、abd-A、Abd-B、bi、svp、disco、Ubx、cad、salm、vvl、pnr、vg、hth、unc-4、trh、H15、Dfd、Rh5、eya。
上述多梳蛋白靶基因的筛选方法，包括以下步骤：
步骤1，在ModENCODE数据库中以“Polycomb group proteins”为关键词检索PcG结合相关的ChIP-seq数据；
步骤2，采用果蝇的Kc167细胞对PcG RNAi之后全基因的表达量进行分析得到RNA-seq数据，再用R语言进行可视化分析，基因表达上调两倍、且P≤0.05，得到PcG targetgenes；
步骤3，将步骤1获取的基因与步骤2获取的基因取交集，获得PcG靶基因。
一种癌症辅助诊断和/或预后判断的试剂盒，包括上述多梳蛋白靶基因。
上述试剂盒，还包括有用于扩增上述多梳蛋白靶基因的引物对。
上述试剂盒在预测癌基因中的应用，是对受试者的病理标本进行权利要求1所述的多梳蛋白靶基因表达水平的检测，基于基因表达水平进行分析评估。
有益效果：本发明通过搜集已有的ChIP-seq数据，结合PcG与靶基因之间的调控关系以及芯片等数据，筛选得到的基因可以作为PcG直接结合并发挥调控功能的基因，这些基因在细胞的增殖分化中起重要作用，同时可以用来筛选潜在的原癌基因。
附图说明
图1为实施例1中381个PcG靶基因的热图；
图2为实施例1中381个PcG靶基因和其它基因上PcG结合峰的对照。
具体实施方式
实施例1
利用如下步骤筛选获得polycomb group proteins(PcG)靶基因：
A、在ModENCODE数据库中搜索PcG的ChIP-seq数据，检索关键词为“Polycombgroupproteins”，再经过一系列的生物信息学分析，共筛选出1311个PcG结合的基因。如表1：
表1：ChIP-seq筛选出的基因
B、采用果蝇的Kc167细胞做RNAi实验。对PcG RNAi之后全基因的表达量进行分析，得到一个RNA-seq数据。通过相关软件如tophat、cufflinks、cuffmerge、cuffdiff等处理RNA-seq数据，用R语言进行可视化分析，基因表达上调两倍，且P≤0.05的，认为是PcGtarget genes，总共筛选到381个基因，并画出热图等，如图1和图2。
C、将步骤A获取的1311个基因和步骤B获得的381个基因取交集，共得到88个基因，如表2。
表2：PcG靶基因
表1和表2所示基因对应的序列可在https://www.ncbi.nlm.nih.gov/gene/上查询。
这些PcG的靶基因组用于预测潜在的癌基因，所述预测是否为癌基因的步骤如下：在患者进行治疗前，对其所取病理标本进行上述靶基因表达水平的检测，分析靶基因是否在病人组织标本中异常表达，基于基因表达水平进行统计学上的聚类分析，确定有哪些可以作为癌基因。
本发明提供了一种在生物样品中进行测定癌基因的试剂盒，所述试剂盒包含：用于检测一组基因的表达水平的试剂，该组基因由以下基因构成：ococ、nub、iab-4、btd、opa、grn、retn、Dll、unpg、mab-21、Kr、disco-r、so、E5、ap、drm、dac、ems、slp2、Vsx1、Ptx1、Oaz、Lim1、Vsx2、dve、rpr、pdm2、lab、pros、wg、peb、Sox100B、tsh、kn、CG14762、Drl-2、CG31778、CG8157、Psc、Sox102F、CG13321、ss、Su(z)2、KP78a、GATAe、klu、KP78b、gsb-n、D、slgA、elB、lz、zfh2、Apt、bab2、CG32121、SoxN、CG10357、Hexo2、mirr、Ama、Zasp52、modSP、fkh、tio、CG33465、CG5613、CG9328、Sr-CIV、Antp、abd-A、Abd-B、bi、svp、disco、Ubx、cad、salm、vvl、pnr、vg、hth、unc-4、trh、H15、Dfd、Rh5、eya。

Claims (5)

1.一组多梳蛋白靶基因，其特征在于：包括ococ、nub、iab-4、btd、opa、grn、retn、Dll、unpg、mab-21、Kr、disco-r、so、E5、ap、drm、dac、ems、slp2、Vsx1、Ptx1、Oaz、Lim1、Vsx2、dve、rpr、pdm2、lab、pros、wg、peb、Sox100B、tsh、kn、CG14762、Drl-2、CG31778、CG8157、Psc、Sox102F、CG13321、ss、Su(z)2、KP78a、GATAe、klu、KP78b、gsb-n、D、slgA、elB、lz、zfh2、Apt、bab2、CG32121、SoxN、CG10357、Hexo2、mirr、Ama、Zasp52、modSP、fkh、tio、CG33465、CG5613、CG9328、Sr-CIV、Antp、abd-A、Abd-B、bi、svp、disco、Ubx、cad、salm、vvl、pnr、vg、hth、unc-4、trh、H15、Dfd、Rh5、eya。
2.权利要求1所述的多梳蛋白靶基因的筛选方法，其特征在于：包括以下步骤：
步骤1，在ModENCODE数据库中以“Polycomb group proteins”为关键词检索PcG结合相关的ChIP-seq数据；
步骤2，采用果蝇的Kc167细胞对PcG RNAi之后全基因的表达量进行分析得到RNA-seq数据，再用R语言进行可视化分析，基因表达上调两倍、且P≤0.05，得到PcG target genes；
步骤3，将步骤1获取的基因与步骤2获取的基因取交集，获得PcG靶基因。
3.一种癌症辅助诊断和/或预后判断的试剂盒，其特征在于：包括权利要求1所述的多梳蛋白靶基因。
4.根据权利要求3所述的癌症辅助诊断和/或预后判断的试剂盒，其特征在于：还包括有用于扩增上述多梳蛋白靶基因的引物对。
5.权利要求3所述的试剂盒在预测癌基因中的应用，其特征在于：对受试者的病理标本进行权利要求1所述的多梳蛋白靶基因表达水平的检测，基于基因表达水平进行分析评估。
CN201711127622.0A 2017-11-15 2017-11-15 一组多梳蛋白靶基因及其筛选方法和应用 Pending CN107868788A (zh)

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