Study About Eye Complication Of Nasopharyngeal Carcinoma And Inhibition Of Gene By Combining SiRNA In Nasopharyngeal Carcinoma Cells

Objective:1.The purpose of clinical study is to explore the eye complication of nasopharyngeral carcinoma(NPC),to analyses the clinical manifestation,CT characteristics and pathological diagnosis of eye complications of NPC,to provide the base for early diagnosis of NPC.2.The purpose of experiment study is to explore the effect of separate and combining hTERTsiRNA,PCNAsiRNA on NPC CNE2 cell;to explore whether targeting siRNA(hTERT, PCNA separate and combining) could inhibit the expressing of related gene and related protein of NPC CNE2 cell,and tumor cells growth,meantime to explore the effect of targeting siRNA (hTERT,PCNA separate and combining ) to cell cycle and apoptosis of NPC CNE2 cell,and to discuss the future application on siRNA in the gene therapy of NPC.Methods:1.The part of clinical study:to retrospective study of 82 cases eye complications in 562 cases NPC,to study their clinical manifestation,CT characteristics and pathological diagnosis.2. The part of experiment study:l) to designing,synthesis and screen effective siRNA ofhTERT and PCNA.2) Effective hTERTsiRNA,PCNAsiRNA separate and combining was transfected into NPC CNE2 cell line by using Lip2000TM siRNA transfection reagent.The relate mRNA and protein were detected by Real-time polymerase chain reaction(Real-time PCR) and immunohistochemistry respectively.MTT assay was used to determine the inhibitory effect of siRNA on tumor cells growth and inverted phase contrast microscope and flow cytometry were used to observe the cell growth,cell cycle,and the apoptosis induced by siRNA.Results:1.The clinical studies showed that eye complication cases were 82cases of 562 NPC cases(14.6%),there were male 64 cases(70eyes) and female 18cases(21 eyes),The mean age at diagnosis was 50.5 years.The bilateral eyes were involved in 9cases.65cases came from Guangdong,6cases from Hunan,4cases from Jiangxi,4cases from Guangxi,2cases from Hainan, 1 cases from Hubei.32cases(37eyes) with blurred vision,30cases(31 eyes) with diplopia, 25cases(26eyes) with ocular movement disorder,12cases(14eyes)with decrease of visual acuity, 8cases with ptosis,6cases with acroisa,3cases with fossa orbitalis swelling,3cases with exophthalmos,1case with ophthalmalgia,36 cases in left and 37 cases in right eyes,9 cases in bilateral eyes.CT appearances:40cases suffered from skull base distracted,6cases with orbit involved,2cases(in left eyes) with orbit metastasis,6cases with sphenoidal sinus involved, 3cases maxillary sinus and 3cases ethmoidal sinus,68case with parapharyngeal space involved, 49cases with soft issue in wall of styloid process involved(there were many kind of shows in th same case,so the data were repeated in these cases).2.The experimental studies showed that 1) Separate and combining siRNA(hTERT,PCNA) transfected could inhibit the expression of hTERT,PCNA and RB1 gene,and cell growth effectually.2) Apoptosis were not observed in transfected CNE2 cells after 48h,but cell cycle stasis in G0/G1 stage.3) Separate and combining siRNA transfected could inhibit the expressio of hTERT,PCNA and RB1 protein obviously.4) Combining siRNA transfected was better than separate siRNA transfected in inhibiting the expression of related gene,protein and cell cycle growth,the difference was significant.Conclusion:1.As there were multiplicity and complexity in eye complication of NPC,so clinician should think highly of these cases.2.Separate and combining siRNA transfected (hTERT,PCNA) could inhibit the expression of related gene and protein of CNE2 cell,and interfere cell growth and cell cycle effectually,combining siRNA transfected was better than separate siRNA transfected in interfering effects to NPC CNE2 cell.This study provides new thoughts and methods for NPC gene therapy.