8种人胰腺癌细胞株常见肿瘤相关基因突变特征分析  

Identification and analysis of tumor-related gene mutations in 8 human pancreatic cancer cell lines by next-generation sequencing

作　　者：王琦 陈凯 金博 杨尹默[1] 田孝东[1] Wang Qi;Chen Kai;Jin Bo;Yang Yinmo;Tian Xiaodong(Department of General Surgery,Peking University First Hospital,Beijing 100034,China;Department of Clinical Laboratory,Peking University First Hospital,Beijing 100034,China)

机构地区：[1]北京大学第一医院普通外科,100034 [2]北京大学第一医院检验科,100034

出　　处：《中华实验外科杂志》2020年第4期667-669,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81572339、81672353、81871954)。

摘　　要：目的分析8种人胰腺癌细胞株中常见肿瘤相关基因突变,为胰腺癌基础研究中选择合适的细胞株提供依据。方法体外培养8种人胰腺癌细胞株(AsPC-1、BxPC-3、Capan-1、Capan-2、MIA PaCa-2、PANC-1、Patu8988和T3M4,均购自美国模式培养物集存库),采用二代测序技术检测113个肿瘤相关基因突变情况,结合京都基因与基因组百科全书(KEGG)数据库对突变基因进行功能分析。结果二代测序结果显示,8种人胰腺癌细胞株中共确定36个基因、79个位点发生突变。鼠类肉瘤病毒癌基因(KRAS)、肿瘤蛋白P53(TP53)、细胞周期依赖性激酶抑制基因(CDKN2A)和胰腺癌缺失基因(DPC4/SMAD4)在8种胰腺癌细胞株中的突变率分别为87.5%(7/8)、100.0%(8/8)、50.0%(4/8)和37.5%(3/8)。AsPC-1和Capan-1细胞中同时存在上述4个基因的突变,BxPC-3细胞株是KRAS野生型胰腺癌,CDKN2A基因在AsPC-1、Capan-1、Capan-2和Patu8988细胞中检测到突变,而DPC4/SMAD4在AsPC-1、Capan-1和T3M4细胞中发生突变。其他发生率较高的突变基因包括AT丰富结合域1A(ARID1A)基因(AsPC-1、Capan-1、Capan-2、MIA PaCa-2和T3M4发生突变)、环磷腺苷效应元件结合蛋白(CREBBP)基因(Capan-1、Capan-2、MIA PaCa-2和T3M4发生突变)、NOTCH1(BxPC-3、Capan-2、MIA PaCa-2和T3M4发生突变)、腺瘤性息肉病(APC)基因(AsPC-1、Capan-1和Patu8988发生突变)和E1A结合蛋白P300(EP300)基因(BxPC-3、MIA PaCa-2和T3M4发生突变)。突变基因功能主要涉及Ras/丝裂原活化蛋白激酶(MAPK)信号通路、细胞周期调控、Wnt/β-连环蛋白(β-catenin)通路、NOTCH信号通路、染色质重塑复合物家族、转化生长因子β(TGF-β)信号通路、DNA损伤修复、Hedgehog和磷脂酰肌醇3激酶丝氨酸/苏氨酸激酶(PI3K-Akt)信号通路等。结论 8种胰腺癌细胞株具备人胰腺癌组织的基因学特征,不同细胞株具有不同的基因突变特点,实验研究中应根据不同细胞的基因突变情况合理选择细胞株�Objective To identify and analyze the characteristics of tumor-associated gene mutations in 8 human pancreatic cancer cell lines,in order to provide a basis for the selection of appropriate cell lines in laboratory research of human pancreatic cancer.Methods Eight human pancreatic cancer cell lines(AsPC-1,BxPC-3,Capan-1,Capan-2,MIA PaCa-2,PANC-1,Patu8988 and T3M4)were purchased from American Type Culture Collection(ATCC)and cultured in vitro,and 113 tumor-associated gene mutations were detected by next-generation sequencing technology.The frequency of mutational genes was calculated,and the functional analysis was performed according to Kyoto Encyclopedia of Genes and Genomes(KEGG)database.Results The next-generation sequencing showed that mutations in 79 loci of 36 genes were identified in 8 human pancreatic cancer cell lines.The mutation rates of Kirsten ratsarcoma viral oncogene homolog(KRAS),tumor antigen p53(TP53),cyclin-dependent kinase inhibitor(CDKN2A)and deleted in pancreatic carcinoma locus 4(DPC4/SMAD4)genes in the 8 cell lines were 87.5%(7/8,except for BxPC-3),100.0%(8/8),50.0%(4/8,in AsPC-1,Capan-1,Capan-2 and Patu8988 cells)and 37.5%(3/8,in AsPC-1,Capan-1and T3M4 cells),respectively.Other mutations with higher incidences included AT rich interactive domain 1A(ARID1A)in AsPC-1,Capan-1,Capan-2,MIA PaCa-2 and T3M4 cells,CREB-binding protein gene(CREBBP)in Capan-1,Capan-2,MIA PaCa-2 and T3M4 cells,NOTCH1 in BxPC-3,Capan-2,MIA PaCa-2and T3M4,adenomatous polyposis coli(APC)in AsPC-1,Capan-1 and Patu8988,and E1A binding protein p300(EP300)in BxPC-3,MIA PaCa-2 and T3M4.The functions of the 36 mutated genes were complex,involving Ras/MAPK signaling pathway,cell cycle transition,Wnt/β-catenin and NOTCH signaling pathway,chromatin remodeling complex family,TGF-βsignaling pathway,DNA damage repair,Hedgehog and PI3K-Akt signaling pathways.Conclusion The 8 human pancreatic cancer cell lines are genetically representative of human pancreatic cancer tissues.Different cell lines have different characteristics of g

关 键 词：胰腺癌 细胞株 基因突变 二代测序 

分 类 号：R735.9[医药卫生—肿瘤]