fancd2 shrna干扰对头颈鳞癌颈淋巴结转移细胞化疗敏感性的影响和机制研究word格式论文.docx

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fancd2 shrna干扰对头颈鳞癌颈淋巴结转移细胞化疗敏感性的影响和机制研究word格式论文.docx

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fancd2 shrna干扰对头颈鳞癌颈淋巴结转移细胞化疗敏感性的影响和机制研究word格式论文

EffectofchemosensitivityandthemechanismsofFANCD2shRNAinterferenceincervicallymphnodemetastasiscellsofheadandnecksquamouscellcarcinomaAbstract:Objective:Theincidenceofheadandnecksquamouscell carcinoma(HNSCCC)rankssixthinsystemictumor.Becauseofthe centralizationofvitalorgansandplentyoflymphnodesintheheadandneck, thelymphnodemetastasisofHNSCCrestrictstheeffectofoperationand radio-chemotherapy.Atpresent,chemotherapyhaswidelyusedinclinical treatmentofHNSCC,especiallythepatientsofcervicallymphnodemetastasis. However,primaryoracquiredresistancetochemotherapyhasdecreasedthe therapeuticeffectontheHNSCCpatientswithlymphnodemetastasis.Fanconi anemia(FA)pathwaydysfunctioncanchangethesensitivityoftumorcellsto DNAcross-linkingagents,andFANCD2asacriticalgeneoftheFApathway mayplayanimportantroleinaffectingthetumorcellgrowthand chemosensitivity.Inthisexperiment,byusingtheHNSCClymphnode metastasisHSC-4 celllineswhichhasstabilityof FANCD2silentexpressionby shRNAinterference,thepossibleroleofFANCD2inHNSCCgrowingandthe effectofsensitivitytochemotherapyandtherelatedmechanismbysilencing FANCD2expressioninHNSCCnecklymphnodemetastasiscellswerestudied. Methods:UsedtheHNSCCnecklymphnodemetastasis HSC-4 cell line whichhadstablyFANCD2silencingeffectbyshRNAinterferenceinearlyexperiment,experimentalcellsweredividedintothreegroups:controlgroup (wild-typeHSC-4cellswithoutanytreatment),negativecontrolgroup (FANCD2-shRNA-C,transfectedinvalidinterferencesequenceofFANCD2in HSC-4cells),experimentalgroup(FANCD2-shRNA,containingeffective interferencesequenceandstablesilencingeffectofFANCD2inHSC-4 cells).Aftercellpassage,puromycinselection,silencingeffectofFANCD2 proteinwasdetectedinWesternBlot,andthesilencingefficiencywasalso calculated.DetectedthegrowthandproliferationofHSC-4cellsby proliferationinhibitionassay(CCK-8method)afterFANCD2shRNA interference,apoptosisandcellcycleofHSC-4cellswasdetectedbyFCM (AnnexinV/PIstainingmethod).Inordertoresearchtheinfluence onthe chemotherapysensitivitybyFANCD2shRNA inH