摘要
目的 筛选胃癌相关血浆差异表达的环状RNA(circRNA)基因,验证初筛circRNA在胃癌患者血浆中的表达,并通过生物信息学方法分析其母基因在胃癌组织中的表达和临床意义.方法 采用circRNA基因芯片技术,获取circRNA在胃癌患者和正常体检者血浆中的表达谱,筛选出胃癌患者与正常体检者之间差异表达2倍以上的circRNA;通过生物信息学方法,筛选候选circRNA基因,分析前10个circRNA对应母基因中在胃癌组织和癌旁组织之间差异表达倍数大于2倍的母基因[睾丸表达10基因(TEX10)、硫酸肝素蛋白多糖2(HSPG2)、微管蛋白ε和δ复合物1(TEDC1)、整合素亚单位β4(ITGB4)、ADP核糖基化因子GTPase激活蛋白1(ARFGAP1)]在胃癌组织中的表达,以及在不同增殖阶段、不同分化程度、有无幽门螺杆菌(Hp)感染、不同淋巴结转移、不同临床病理组织亚型患者胃癌组织与癌旁正常组织间的差异表达.结果 通过分析胃癌患者血浆相关circRNA表达谱,共发现566种circRNA在胃癌患者和正常体检者血浆中的表达水平存在显著性差异(P＜0.05),其中68种circRNA表达水平差异超过2倍.通过生物信息学分析,最终筛选得到20个候选circRNA基因.母基因TEX10、HSPG2、TEDC1、ITGB4、ARFGAP1在胃癌组织中明显上调表达(P＜0.001),在不同增殖阶段、不同分化程度、有无Hp感染、不同淋巴结转移、不同临床病理组织亚型患者胃癌组织与癌旁正常组织中的表达均具有显著性差异(P＜0.05或P＜0.01或P＜0.001).结论 成功构建了胃癌相关血浆circRNA表达谱,并初步筛选出在胃癌患者血浆中差异表达的20个circRNA基因;血浆中升高的 hsa_circ_0087773、hsa_circ_0010697、hsa_circ_0033634、hsa_circ_0045686、hsa_circ_0061150 所对应的母基因TEX10、HSPG2、TEDC1、ITGB4、ARFGAP1在胃癌组织中亦上调表达,这对胃癌的诊断和预后评估具有一定的临床价值.
Abstract
Objective To screen the plasma differentially expressed circular RNA(circRNA)genes related to gastric cancer,verify the expression of initial screening circRNA in plasma of patients with gastric cancer,and ana-lyze the expression and clinical significance of their parent genes in gastric cancer tissues by bioinformatics methods.Methods CircRNA gene chip technology was used to obtain the expression profile of circRNA in the plasma of gastric cancer patients and normal physical examination subjects,and the circRNA whose expression difference was more than 2 times between gastric cancer patients and normal physical examination subjects was selected.Candidate circRNA genes were screened by bioinformatics methods.The parent genes corresponding to the top 10 circRNAs that were differentially expressed by a factor greater than 2 between gastric cancer tissues and adjacent tissues were analyzed[testis expressed 10(TEX10),heparin sulfate proteoglycan 2(HSPG2),tubulin e and 8 complex 1(TEDC1),integrin subunit β4(ITGB4),ADP ribosylation factor GTPase activating protein 1(ARFGAP1)]in gas-tric cancer tissues.And the differential expression between gastric cancer tissues and adjacent normal tissues in pa-tients with different proliferation stages,different differentiation degrees,helicobacter pylori(Hp)infection or not,different lymph node metastasis,and different clinicopathological subtypes was analyzed.Results By analyzing the expression profiles of circRNA in the plasma of gastric cancer patients,it was found that there were significant diffe-rences in the expression levels of 566 circrRNAs in the plasma of gastric cancer patients and normal physical exami-nation subjects(P＜0.05),and the expression levels of 68 circRNAs were more than 2 times different.Through bioin-formatics analysis,20 candidate circRNA genes were finally screened.The expressions of parent genes TEX10,HSPG2,TEDC1,ITGB4 and ARFGAP1 were significantly up-regulated in gastric cancer tissues(P＜0.001).There were significant differences in expression between gastric cancer tissues and adjacent normal tissues in patients with different proliferation stages,different differentiation degrees,Hp infection or not,different lymph node metastasis,and different clinicopathological subtypes(P＜0.05 or P＜0.01 or P＜0.001).Conclusion The circRNA expression profile related to gastric cancer was successfully constructed,and 20 differentially expressed circRNA genes were initially screened out in the plasma of gastric cancer patients.The parent genes TEX10,HSPG2,TEDC1,ITGB4,and ARFGAP1 corresponded to highly expressive hsa_circ_0087773,hsa_circ_0010697,hsa_circ_0033634,hsa_circ_0045686,and hsa_circ_0061150 in plasma were also up-regulated in gastric cancer tissues,which has certain clinical value for the diagnosis and prognosis evaluation of gastric cancer.
基金项目
甘肃省卫生及计划生育委员会项目(GSWSKY2017-15)
兰州市科技发展计划(2017-4-83)
云南省教育厅科研项目(2019J1244)
甘肃省人民医院国家级科研项目培育计划(19SYPYB-25)
国家科技期刊平台
NSTL
万方数据