The Research Of The Relation Between Serum MicroRNA-329 Level And Development Of Nasopharyngeal Carcinoma

Objective To explore the serum levels of microRNA-329 and the correlation of poor prognosis in patients with nasopharyngeal carcinoma and the occurrence and development of microRNA-329 control nasopharyngeal carcinoma specific molecular mechanism is studied.Methods Nasopharyngeal carcinoma(NPC)is the most common malignant tumor of head and neck in the southern China,it is sensitive to radiation,due to the anatomical position ofnasopharynx is concealed,early patients usually treated with radiotherapy,while in patients with advanced usually need to be treated with radio therapy and adjuvant chemotherapy,the five years overall survival rate of patients is usually about 70%,and the five years recurrence and distant metastasis rate of patients is about 3 0%-40%.Recurrence anddistant metastasis of nasopharyngeal carcinoma impact on the survival and health and quality of life of patients,and the nasopharyngeal carcinoma tumor markers are low sensitivity and specificity,so we need to explore serologistic biomarkers which with high sensitivity and specificity, easy to dynamic observation so that to strengthen monitoring and early intervention on recurrence and metastasis of patients.microRNAcoRNAs(microRNANAs)is a single stranded,small non coding RNA molecules with regulated function,which length is about 18～25nt.It usually combined with its target molecule mRNA 3’noncoding region(3’-UTR),and inhibit the translation of target gene,participate in the process of life.Although the biologistic function of microRNANAs is notclear,but it is certain that they have relatively specific expression pattern,also participate invarious pathologistic and physiologistic process of life.Abnormal expression of microRNANAs arerelated to human diseases,in particularly,a large number of studies have shown,microRNANAs is a key molecule involved in the tumorigenesis and development of common tumors,such as colorectal cancer,breast cancer,lung cancer,esophageal cancel ovarian cancer.microRNANAs is considered to be oncogenes or arnti-oncogenes and it have specific spectrum.It is worth to emphasize that circulating microRNANAs are relatively stable and easy to bedetected,and can be used for dynamic observation of tumor and evaluate the clinical analysis and the prognosis of the patients,therefore,as a new tumor marker,circulating microRNANA has good prospect of clinical application.First of all,the first part we study from the level of clinical expression level of serum microRNA-329 in patients with nasopharyngeal carcinoma and explore the relevance and the related clinical index.In our October 2014 to October 2014 in our hospital for treatment of 200 cases of patients with nasopharyngeal carcinoma acquisition of serum and collect relevant clinical data of patients,as the team;In October 2014 to October 2016 in our hospital during the period of physical examination of 200 cases of normal check-up crowd serum are collected,as normal control group.Using microRNAnas chip technology to research in patients with nasopharyngeal carcinoma and the control group of normal physical examination crowd serum microRNAnas level testing,the results showed that serum microRNA-329 levels in patients with nasopharyngeal carcinoma present a significantly lower expression.Through single factor analysis and multifactor logistic non conditional analysis way of microRNA-329 test and clinical indicators of patients,the expression of microRNA-329 levels with the patient’s tumor metastasis,recurrence and survival rate of clinical indexes such as correlation analysis.This study as well as the organization,to detect the expression of microRNA-329 in compared with serum microRNA-329 levels,found that there is obvious positive correlation.The second part we study from the level of molecular biology,cell of microRNA-329 control explore the molecular mechanism of nasopharyngeal carcinoma development.We adopt western blot analysis technology,real-time fluorescent quantitative PCR technology,immune cell fluorescence staining techniques,determination of growth curve,transwell,fluorescent report enzymes as well as the experimental technology of microRNA-329 impact on the ability of cell growth and metastasis of nasopharyngeal carcinoma and explore the molecular mechanism.Our third part of the research from the level of molecular biology,animal of microRNA-329 control to verify the molecular mechanism of nasopharyngeal carcinoma development.using nude mice into tumor,tail vein injection technology and other technical of microRNA-329 impact on the ability of cell growth and metastasis of nasopharyngeal carcinoma(and its molecular mechanism for validation.Results The first part of microRNAnas chip results showed that compared with normal physical examination,serum microRNA-329 levels in patients with nasopharyngeal carcinoma is obviously lower expression.Nasopharyngeal carcinoma was associated with a significant reduction in the level of serum microRNA-329,and the levels of serum microRNA-329 recurrence and metastasis rate in patients with nasopharyngeal carcinoma,a significant negative correlation,and is related to the survival rate of patients with nasopharyngeal carcinoma.microRNA-329 levels in patients with nasopharyngeal carcinoma consistent with the level of serum microRNA-329,showing significant positive correlation.The second part of results showed that inhibition of microRNA-329 in nasopharyngeal carcinoma cells,cell transfer ability increases significantly,a marked increase in the ability of cell growth;Overexpression of microRNA-329 in nasopharyngeal carcinoma cells,cell transfer ability declined significantly,cell proliferation ability to rise.And enzyme fluorescent report experimental results show that microRNA-329 can by acting on target genes Wnt-7B,realize the inhibition of Wnt-7 bmrna level,thus inhibiting B-catenin into nuclear,inhibition of Wnt signaling pathway,so as to realize the inhibition of cell growth and metastasis ability.The third part of results showed that at least a tumor experiment results show that the cells compared with control group,the expression of microRNA-329 nasopharyngeal carcinoma cells into a tumor is more slow,into tumors had smaller,into tumor sexual difference;Tail intravenous injection into the tumor experimental results showed that compared with the control cells,expression of microRNA-329 nasopharyngeal carcinoma cells transfer ability dropped significantly.Conclusion The low expression level of serum microRNA-329 was closely related to poor prognosis in patients with nasopharyngeal carcinoma,microRNA-329 is by acting on Wnt-7 b to inhibition of Wnt signaling pathways,thereby inhibit the growth of nasopharyngeal carcinoma cells and transfer skills.Serum microRNA-329 can be used as a marker to predict the prognosis of patients with nasopharyngeal carcinoma effect,microRNA-329 and its target gene can be used as potential therapeutic targets of nasopharyngeal carcinoma.