Screen And Identification Of Specially Expressed Micrornas In Nasopharyngeal Carcinoma

Objective: The chip technology analysis nasopharyngeal carcinomapatients and normal serum miRNA expression difference spectrum, screeningnasopharyngeal carcinoma, and specific serum miRNA to screening miRNAverification, looking for difference expression of serum peptide and serummiRNA for nasopharyngeal carcinoma, to early diagnosis and conditionalmonitoring and prognosis of the disease providing the basis and theoreticalfoundation, finding the diagnosis and treatment of nasopharyngeal carcinoma(NPC) new ideas.Methods:Part I Screening of Differentially expressed miRNA in NPCpatients with miRNAs chips（1）According to the serum collection standard operating procedures ofthe collection of nasopharyngeal carcinoma patients serum and normalserum specimens. All of the patients and volunteers are informedconsent and sign informed consent;（2）Using RNAextraction kit extraction patients and volunteers RNA; （3）Using spectrophotometer measurement of RNAquality；（4）Choosing miRCURYTM Array Power kit preparation fluorescentmarker probe, and chip hybridization and chip signal scan;（5）Using GenePix4000b chip scanning fluorescence intensity, withGenePix pro v5.5software for data processing and analysis;Results:（1）Collectting a certain amount of establishing affiliated hospital ofmedical school of guilin in June2011-February2012clinical pathologydiagnosed nasopharyngeal low differentiated squamous cell carcinomain cancer patients and volunteers；（2）The extraction of RNA purity and quality anaslying: extract RNAspecimen optical density measured value A260/A280greater than1.7,indicating that extract serum RNA high purity, RNA quantity is morethan10ng conform to the requirements of the gene chip to specimenNasopharyngeal carcinoma；（3）It was found that54serum miRNAs in NPC patients were two foldhigher than that in the normal, and22miRNAs were five times higherthan the normal. While88serum miRNAs in NPC patients were twotimes than the normal, and the number of miRNAs five times lower thanthe normal was16.（4）The serum miRNAs related EBV in NPC patients that were twotimes higher than the normal included EBV-miR-BART4,EBV-miR-BART18-3p,EBV-miR-BART19-5andEBV-miR-BART21-3p. Part II RT-PCR identification of serum miRNAs of high expression inNPC patients that were related EBV and screened bymicRNAs chips.（1）miRNA were extracted from nasopharyngeal carcinoma andnasopharyngeal mucosa inflammation tissues.（2）real-time fluorescent quantitative PCR reaction.（3）miRNA were extracted from the serum of NPC patients andnasopharyngeal mucosa inflammation patients.（4）The reverse transcription of miRNAs.Results:Serum miRNAs might be closely related with the incidence ofnasopharyngeal carcinoma.Conclusions:（1）It was found that54serum miRNAs in NPC patients were two foldhigher than that in the normal, and22miRNAs were five timeshigher than the normal. While88serum miRNAs in NPC patientswere two times than the normal, and the number of miRNAs fivetimes lower than the normal was16.（2）The serum miRNAs related EBV in NPC patients that were two timeshigher than the normal included EBV-miR-BART4，EBV-miR-BART18-3p,EBV-miR-BART19-5andEBV-miR-BART21-3p.（3）Serum miRNAs might be closely related with the incidence ofnasopharyngeal carcinoma.