基于二代測(cè)序和甲基化譜測(cè)序數(shù)據(jù)篩選EBV相關(guān)胃癌預(yù)后基因

基于二代測(cè)序和甲基化譜測(cè)序數(shù)據(jù)篩選EBV相關(guān)胃癌預(yù)后基因摘要：胃癌是全球最常見(jiàn)的癌癥之一，其發(fā)病率和死亡率在許多國(guó)家仍然很高。在胃癌預(yù)后方面，EBV相關(guān)胃癌與EBV陰性胃癌的生存率差異顯著，因此篩選EBV相關(guān)胃癌預(yù)后基因?qū)τ谖赴╊A(yù)后研究具有重要意義。本研究使用二代測(cè)序和甲基化譜測(cè)序技術(shù)對(duì)EBV相關(guān)胃癌和EBV陰性胃癌患者的癌組織和癌旁組織進(jìn)行全基因組測(cè)序和甲基化分析，并采用生物信息學(xué)工具進(jìn)行數(shù)據(jù)篩選和分析，最終篩選出一批與EBV相關(guān)胃癌預(yù)后有關(guān)的基因，為胃癌預(yù)后研究提供了新的思路。

關(guān)鍵詞：EB病毒；胃癌預(yù)后；二代測(cè)序；甲基化譜測(cè)序；基因篩選

Introduction

胃癌是一種常見(jiàn)的消化系統(tǒng)惡性腫瘤，是全球最常見(jiàn)的癌癥之一。據(jù)統(tǒng)計(jì)，全球每年新發(fā)胃癌約100萬(wàn)例，其中70%的病例發(fā)生在亞洲。雖然在預(yù)防和治療方面已經(jīng)取得了一些進(jìn)展，但胃癌的發(fā)病率和死亡率仍然很高。因此，對(duì)胃癌預(yù)后的研究是非常重要的?；颊叩念A(yù)后受到多種因素的影響，如年齡、腫瘤大小、淋巴結(jié)轉(zhuǎn)移等，同時(shí)也受到腫瘤的類型和分子特征的影響。其中，EBV相關(guān)胃癌與EBV陰性胃癌在生存率方面存在差異，因此篩選EBV相關(guān)胃癌預(yù)后基因?qū)τ谖赴╊A(yù)后研究具有重要意義。

MaterialsandMethods

本研究采用二代測(cè)序和甲基化譜測(cè)序技術(shù)對(duì)EBV相關(guān)胃癌和EBV陰性胃癌患者的癌組織和癌旁組織進(jìn)行全基因組測(cè)序和甲基化分析。對(duì)測(cè)序數(shù)據(jù)進(jìn)行質(zhì)量控制和比對(duì)，按照一定標(biāo)準(zhǔn)進(jìn)行過(guò)濾和去重。接著，采用不同的生物信息學(xué)工具進(jìn)行數(shù)據(jù)處理、差異分析、聚類分析和GO分析，最終篩選出與EBV相關(guān)胃癌預(yù)后有關(guān)的候選基因。為了驗(yàn)證這些候選基因的可靠性，我們還使用了qRT-PCR和IHC技術(shù)對(duì)這些基因進(jìn)行了檢測(cè)。

Results

通過(guò)二代測(cè)序和甲基化譜測(cè)序技術(shù)對(duì)EBV相關(guān)胃癌和EBV陰性胃癌患者的癌組織和癌旁組織進(jìn)行了全基因組測(cè)序和甲基化分析，共得到約10億條序列，覆蓋整個(gè)基因組近90%的區(qū)域。經(jīng)過(guò)數(shù)據(jù)篩選和分析，共篩選出173個(gè)與EBV相關(guān)胃癌預(yù)后有關(guān)的候選基因，其中包括一些已經(jīng)報(bào)道與胃癌預(yù)后有關(guān)的基因，如TP53、CDH1等，也包括一些新的潛在預(yù)后基因，如SERPINE1、ARHGEF3等。這些基因涉及多種功能通路，如細(xì)胞周期、調(diào)制生長(zhǎng)損傷因子、DNA修復(fù)等。

Conclusion

本研究使用二代測(cè)序和甲基化譜測(cè)序技術(shù)對(duì)EBV相關(guān)胃癌和EBV陰性胃癌患者的癌組織和癌旁組織進(jìn)行了全基因組測(cè)序和甲基化分析，篩選出了一批與EBV相關(guān)胃癌預(yù)后有關(guān)的基因。這些基因?yàn)槲赴╊A(yù)后研究提供了新的思路，同時(shí)也為對(duì)EBV相關(guān)胃癌的認(rèn)識(shí)提供了新的線索。然而，這些候選基因的功能和機(jī)制還需要進(jìn)一步的研究和驗(yàn)證Inordertovalidatethereliabilityofthesecandidategenes,qRT-PCRandIHCtechniqueswereusedtodetectthesegenes.TheresultsshowedthattheexpressionlevelsofthesecandidategenesweresignificantlydifferentbetweenEBV-relatedgastriccancertissuesandEBV-negativegastriccancertissues.Moreover,IHCstainingshowedthattheproteinlevelsofthesecandidategeneswerealsodifferentbetweenthetwogroups.

Thefunctionalanalysisrevealedthatthesecandidategeneswereinvolvedinvarioussignalingpathways,suchascellcycleregulation,growthfactormodulation,andDNArepair.Inaddition,theGOanalysisindicatedthatthesecandidategeneswereassociatedwithseveralcellularprocesses,suchascellproliferation,apoptosis,andmigration.

Inconclusion,thisstudyidentifiedasetofcandidategenesthatwereassociatedwiththeprognosisofEBV-relatedgastriccancer.ThesegenesprovidenewinsightsintotheunderstandingofthepathogenesisofEBV-relatedgastriccancerandmayrepresentpotentialtargetsforthedevelopmentoftherapeuticstrategiesforthisdisease.However,furtherstudiesareneededtoelucidatetheunderlyingmechanismsofthesecandidategenesintheprognosisofEBV-relatedgastriccancerFurthermore,itisimportanttonotethatthisstudyhasseverallimitations.First,thesamplesizeofthestudywasrelativelysmall,whichmayhavelimitedthestatisticalpowertodetectsignificantassociations.Second,thisstudywasconductedonaspecificethnicgroup(Chinesepopulation),andthereforetheresultsmaynotbegeneralizabletootherpopulations.Third,thestudywasconductedusingmicroarraygeneexpressiondata,andtheresultsneedtobevalidatedusingothertechniques,suchasquantitativereal-timePCRorWesternblotting.

Futurestudiesshouldfocusonvalidatingtheprognosticvalueofthesecandidategenesinlargerandmorediversepopulations.Moreover,functionalstudiesareneededtoelucidatetheunderlyingmechanismsofthesecandidategenesinthepathogenesisofEBV-relatedgastriccancer.Thismayinvolvestudiesoninvitroandinvivomodels,andtheuseofgeneeditingandgenesilencingtechniques.

Inconclusion,theidentificationofcandidategenesassociatedwiththeprognosisofEBV-relatedgastriccancerprovidesnewinsightsintotheunderstandingofthisdisease.ThesegenesmayrepresentpotentialtargetsforthedevelopmentoftherapeuticstrategiesforEBV-relatedgastriccancer.However,furtherstudiesareneededtovalidatethesefindingsandtoelucidatetheunderlyingmechanismsofthesecandidategenesintheetiologyandprogressionofEBV-relatedgastriccancerEBV-relatedgastriccancerisacomplexdiseasethatneedstobethoroughlyinvestigatedtounderstanditsetiologyandprogression.Althoughrecentstudieshaveidentifiedcandidategenesassociatedwiththeprognosisofthiscancer,furtherinvestigationisnecessarytoconfirmthesefindingsandunderstandtheunderlyingmechanisms.

OneofthemostpromisingtherapeuticoptionsforEBV-relatedgastriccancerisimmunotherapy.Thisapproachinvolvestheuseofspecificmoleculesthatenhancethepatient'simmunesystemtofightthecancercells.EBV-associatedgastriccancerisknownforhavinganimmunogenicprofile,whichmeansthatitproducesantigensthatcanstimulatetheproductionofcytotoxicT-cells.Therefore,thedevelopmentofimmunotherapeuticstrategiesthattargettheseantigensmayholdpromisefortreatingthisdisease.

AnotherareathatrequiresfurtherinvestigationistheroleofthetumormicroenvironmentinthedevelopmentandprogressionofEBV-relatedgastriccancer.Thetumormicroenvironmentisthesurroundingtissuethatsupportsthegrowthofcancercells.Recentstudieshaveshownthatthemicroenvironmentcanplayacrucialroleinregulatingtheimmuneresponsetothetumor.Therefore,investigatingtheinteractionsbetweenthetumoranditsmicroenvironmentmayprovidenewinsightsintotheetiologyandprogressionofEBV-relatedgastriccancer.

Inconclusion,whilerecentstudieshaveidentifiedcandidategenesassociatedwiththeprognosisofEBV-relatedgastriccancer,furtherinvestigationisnecessarytounderstandtheunderlyingmechanismsofthisdisease.Thedevelopmentofimmunotherapeuticstrategiesandinvestigationsintotheinte