The Differential Expressing MicroRNA Profile Of Lingual Mucosa Carcinogenesis Induced By By4-Nitro Quinoline1-Oxide In C57/BL6Mice

[Object] By studying the difference expression of miRNAs in the process of tongue mucosa cancer, explore the molecular pathological mechanism of oral cancer, to look for hunman oral cancer early diagnosis, prognosis of specific molecular markers, and provide objective basis for new therapeutic targets.[Method]1.85female C57/BL6mice are divided into groups a randomly,the fist group which includes5mice drink water normally,the second group which includes75mice to drink water contained4NQ0, the third group which includes5mice drink solvents propylene glycol water,2nd,3rd groups after16weeks are given normal water.1st group:blank control, at week28put to death;2nd group:after the official start of the experiment,5mice are put to death, and every two weeks were randomly put to5mice death,until28week.3rd groups:propylene glycol solvent comparison, put to death at28week.2.To divide the mice tongue into two parts uniformly from the middle line, one half is covered in RNAlater and kept4°C refrigerator for24hours, separation membrane, the membrane is covered in new RNA later again,-80℃preservation; the other one is soaked in10%formaldehyde solution, from0,4,8,12,16,20,24,28weeks and control group of distilled water,1,2propanediol,randomly selecte three cases and make paraffin section for histopathologic analysis.3. Refer to the histopathological results of1st group,2nd group,3rd groups,select normal tissues, precancerous tissue, mucous membrane of the squamous carcinoma tissues and extract RNA, application of miRNA microarray technology and data analysis, early screening of differentially expressed miRNAs in tongue cancer development. [Result] Reference histopathological results and statistical analysis,which select week0, week12and week28,each group three sample are used to miRNA expression profile chip test:1. compared with week12of tongue mucosa and week0tongue mucosa, expression changes of miRNA molecules have42, which7are upregulation (miR-31-5p, miR-491-5p, miR-672-5p...),34are downregulation(miR-719, miR-M23-1-5p, miR-149-5p...).2. compared with week28mice tongue mucosa and week12of tongue mucosa, expression changes of microRNAs have8, which5are regulation (miR-714, miR-5099, miR-471-5p...),3are downregulation (miR-491-5p, miR-3060-3p, miR-3097-3p).3. compared with week28tongue mucosa in mice and week0mice tongue mucosa,there are42expression changes of microRNAs molecules, which12are upregultion,(miR-31-5p, miR-290-a-3p, miR-688...),30are downregulation (miR-34c-3p, miR-350-3p,miR-138-1-3p...)[Conclusion] there are significant differences of the expression of microRNAs in normal tissue, the mucosa precancerous lesions and squamous carcinoma.This differences implicate complecated molecular regulation mechanism in mucosal canceration, this mechanism still needs further research.