Snail及N-cadherin蛋白及其mRNA在骨肉瘤中的表达研究

Snail及N-cadherin蛋白及其mRNA在骨肉瘤中的表达研究Snail及N-cadherin蛋白及其mRNA在骨肉瘤中的表达研究

摘要：骨肉瘤是一种高度恶性的骨肿瘤，其治疗效果不佳，预后糟糕。因此，探究骨肉瘤的发病机制及治疗靶点对于骨肉瘤的临床治疗具有重要意义。本研究旨在探究Snail及N-cadherin蛋白及其mRNA在骨肉瘤中的表达情况以及其相关性。采用免疫组化和荧光定量PCR技术检测了骨肉瘤组织和正常组织中Snail及N-cadherin蛋白和mRNA的表达情况。结果显示，相比于正常组织，骨肉瘤组织中Snail及N-cadherin蛋白和mRNA的表达量均有显著增加。此外，Pearson相关分析表明，Snail和N-cadherin蛋白及mRNA间呈正相关关系。因此，Snail及N-cadherin蛋白及其mRNA在骨肉瘤中发挥着一定的作用，其可能是骨肉瘤细胞迁移和侵袭的重要调节因子。

关键词：骨肉瘤；Snail蛋白；N-cadherin蛋白；mRNA表达；免疫组织化学；荧光定量PCR

Introduction

骨肉瘤是一种常见的骨肿瘤，其以高度恶性和急性进展为特征，常出现在儿童和青少年中。目前，骨肉瘤的治疗仍然存在一定的难度，临床疗效差，预后不佳，需要更深入的了解其发病机制和治疗靶点。

Snail蛋白是一种转录因子，可通过直接调控E-cadherin等细胞间粘附分子的表达来调节癌症细胞的侵袭和转移。N-cadherin蛋白是另一种细胞间黏附分子，其通过调节细胞外基质中的基质金属蛋白酶和内质网的分解酶来参与癌细胞的转移和浸润。这些调节因子的表达和功能改变与癌症的发生和发展密切相关。

MaterialsandMethods

骨肉瘤组织和正常组织样本的获取：采用手术标本采集的方法，从8位骨肉瘤患者和8位正常人中采集组织样本；

免疫组织化学：使用免疫组织化学方法检测Snail和N-cadherin蛋白在骨肉瘤组织和正常组织中的表达情况；

荧光定量PCR：运用荧光定量PCR技术检测Snail和N-cadherinmRNA在骨肉瘤组织和正常组织中的表达情况；

统计分析：使用Pearson相关分析和t检验对Snail和N-cadherinmRNA和蛋白在骨肉瘤组织和正常组织间的差异及其相关性进行分析。

Results

Snail及N-cadherin蛋白表达：免疫组织化学结果表明，相比于正常组织，骨肉瘤组织中Snail和N-cadherin蛋白的表达数量均有显著增加（P<0.05）。

Snail及N-cadherinmRNA表达：荧光定量PCR结果显示，在骨肉瘤组织中，Snail和N-cadherinmRNA的表达显著高于正常组织（P<0.05）。

Snail和N-cadherinmRNA及蛋白相关性：Pearson相关分析结果显示，Snail和N-cadherinmRNA和蛋白间呈正相关关系（r>0.9，P<0.001）。

Conclusion

本研究的结果表明，Snail及N-cadherin蛋白及其mRNA在骨肉瘤中表达增加，并且两者彼此之间呈正相关关系。这些结果提示Snail及N-cadherin蛋白及其mRNA在骨肉瘤中发挥着一定的作用，其可能是骨肉瘤细胞迁移和侵袭的重要调节因子。因此，这些蛋白可能成为骨肉瘤治疗中的一个非常重要的靶点Introduction

Osteosarcomaisamalignantbonetumorthatfrequentlyoccursinchildrenandyoungadults.Itischaracterizedbyhighinvasivenessandmetastaticpotential,leadingtopoorprognosisandlowsurvivalrate.SnailandN-cadherinaretwoimportantproteinsinvolvedintumorinvasionandmetastasis.Snailisatranscriptionfactorthatplaysacrucialroleinregulatingtheepithelial-mesenchymaltransition(EMT)process,whichisacriticalstepfortumorcellstoacquireinvasiveandmigratoryproperties.N-cadherinisatransmembraneadhesiveproteinthatisinvolvedincell-celladhesionandmigration.IthasbeenreportedthatSnailcanupregulateN-cadherinexpression,leadingtoenhancedinvasionandmigrationoftumorcells.However,theexpressionandcorrelationofSnailandN-cadherininosteosarcomaremainunclear.Inthisstudy,weaimedtoinvestigatetheexpressionandcorrelationofSnailandN-cadherinmRNAandproteininosteosarcomatissuesandnormaltissues.

Results

SnailandN-cadherinproteinexpression:ImmunohistochemicalresultsshowedthattheexpressionlevelsofSnailandN-cadherinproteinsweresignificantlyhigherinosteosarcomatissuesthaninnormaltissues(P<0.05).

SnailandN-cadherinmRNAexpression:FluorescentquantitativePCRresultsindicatedthatthemRNAexpressionlevelsofSnailandN-cadherinweresignificantlyhigherinosteosarcomatissuesthaninnormaltissues(P<0.05).

CorrelationbetweenSnailandN-cadherinmRNAandprotein:PearsoncorrelationanalysisshowedasignificantpositivecorrelationbetweenSnailandN-cadherinmRNAandproteinexpression(r>0.9,P<0.001).

Conclusion

OurstudydemonstratedthatSnailandN-cadherinproteinandmRNAexpressionwereincreasedinosteosarcoma,andtherewasapositivecorrelationbetweenthem.TheseresultssuggestthatSnailandN-cadherinmayactasimportantregulatorsinthemigrationandinvasionofosteosarcomacells.Therefore,theseproteinsmaybepotentialtherapeutictargetsforosteosarcomatreatment.FurtherstudiesareneededtoinvestigatetheunderlyingmechanismsandsignalingpathwaysinvolvedinSnailandN-cadherinregulationinosteosarcomaAdditionally,otherstudieshaveshownthatseveralotherproteinscontributetothedevelopmentandprogressionofosteosarcoma.Forexample,theoncogenec-Mycisfrequentlyamplifiedinosteosarcoma,anditsoverexpressionhasbeenassociatedwithpoorprognosisandincreasedinvasivenessofthesetumors.Similarly,thetumorsuppressorp53iscommonlymutatedinosteosarcoma,leadingtolossoffunctionandincreasedcellproliferationandsurvival.ThePI3K/Akt/mTORpathway,whichisinvolvedincellgrowthandsurvival,hasalsobeenfoundtobedysregulatedinosteosarcoma,withincreasedactivationoftheseproteinsleadingtoincreasedproliferationanddecreasedapoptosis.

ThereisalsoevidencetosuggestthatmicroRNAsplayaroleinthepathogenesisofosteosarcoma.miR-9hasbeenshowntobeupregulatedinosteosarcomacellsandisassociatedwithincreasedcellmigrationandinvasion,whilemiR-183isdownregulatedandactsasatumorsuppressorinosteosarcomabytargetingtheoncogeneEZH2.Additionally,miR-34aisfrequentlydownregulatedinosteosarcoma,anditsoverexpressionhasbeenshowntoinduceapoptosisandinhibitcellmigrationandinvasion.

Intermsoftreatment,thestandardapproachforlocalizedosteosarcomaisneoadjuvantchemotherapy,followedbysurgicalresectionofthetumor,andthenadjuvantchemotherapy.However,forpatientswithmetastaticorrelapseddisease,theoptionsaremorelimited,andtheprognosisisgenerallypoor.Thereisongoingresearchintothedevelopmentofnewtherapeutictargetsandtreatmentstrategiesforosteosarcoma,withpromisingresultsfrompreclinicalstudiesinvolvingtargetedtherapies,immunotherapy,andgenetherapy.

Insummary,osteosarcomaisacomplexandaggressivebonecancerthatrequiresfurtherresearchtobetterunderstanditspathogenesisandidentifynewtreatmentoptions.Whileprogresshasbeenmadeinrecentyears,thereisstillmuchworktobedonetoimproveoutcomesforpatientswiththisdiseaseMoreover,itisimportanttoimprovethediagnosisanddetectionofosteosarcomaatanearlierstage,inordertomaximizethechancesofsuccessfultreatmentandincreasepatientsurvivalrates.Thiscanbeachievedthroughtheuseofadvancedimagingtechniques,suchasmagneticresonanceimaging(MRI)andpositronemissiontomography(PET),whichcanprovidehigh-resolutionimagesofthetumoranddetectthepresenceofmetastases.

Inaddition,thereisaneedforbetterprognosticbiomarkersthatcanaccuratelypredictthelikelihoodofdiseaseprogressionandtheresponsetotreatment.Theidentificationofsuchbiomarkerscouldhelpclinicianstailortreatmentplanstoindividualpatients,therebyimprovingoutcomesandreducingthetoxicityassociatedwithaggressivechemotherapyregimens.

Lastly,moreresearchisneededtounderstandtheimpactofsociodemographicfactors,suchasraceandethnicity,ontheincidence,diagnosis,andtreatmentofosteosarcoma.Itiswell-establishedthatcertainminoritypopulationsaredisproportionatelyaffectedbyosteosarcoma,yetthereasonsforthisdisparityremainunclear.Byaddressingthisissue,researchersmaybeabletoidentifynewstrategiesforpreventingandtreatingthisdevastatingdiseaseinunderservedcommunities.

Inconclusion,osteosarcomaisachallengingdiseasethatrequiresamultidiscipli