| Background and Objectives:Nasopharyngeal carcinoma（NPC） is the most common malignant tumor of head andneck 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 patientswith advanced usually need to be treated with radiotherapy and adjuvant chemotherapy[1],the five years overall survival rate of patients is usually about70%[2], and the five yearsrecurrence and distant metastasis rate of patients is about30%-40%[3]. Recurrence anddistant metastasis of nasopharyngeal carcinoma impact on the survival and health andquality of life of patients, and the nasopharyngeal carcinoma tumor markers are lowsensitivity and specificity[4], so we need to explore serological biomarkers which with highsensitivity and specificity, easy to dynamic observation so that to strengthen monitoring andearly intervention on recurrence and metastasis of patients.MircoRNAs (miRNAs) is a single stranded, small non coding RNA molecules withregulated function,which length is about18~25nt.It usually combined with its targetmolecule mRNA3’noncoding region (3’-UTR), and inhibit the translation of target gene，participate in the process of life[5][6][7]. Although the biological function of miRNAs is notclear, but it is certain that they have relatively specific expression pattern, also participate invarious pathological and physiological process of life. Abnormal expression of miRNAs are related to human diseases, in particularly, a large number of studies have shown, miRNAsis a key molecule involved in the tumorigenesis and development of common tumors,suchas colorectal cancer, breast cancer, lung cancer, esophageal cance， ovarian cancer[1][2].MiRNAs is considered to be oncogenes or anti-oncogenes and it have specific spectrum[3].It is worth to emphasize that circulating miRNAs are relatively stable and easy to bedetected,and can be used for dynamic observation of tumor and evaluate the clinicalanalysis and the prognosis of the patients, therefore, as a new tumor marker, circulatingmiRNA has good prospect of clinical application[4][5][6].Ki-67is markers of cell proliferation state,usually is closely related to thepathological process of cancer[7].As the soluble fragments of cytokeratin19（CK-19）,serum Cyfra21-1is especially rich in squamous cell carcinoma[8].Serum squamous cellcarcinoma antigen (SCCA) is a serological markers of squamous cell carcinoma[9].TheLDH and ALP are biochemistry markers, and are also reported associated with cancertumorigenesis[10][11].But the combined detection of serum tumor markers and miRNAswere used to study the monitoring of recurrence and distant metastasis of nasopharyngealcarcinoma has not been reported.With the application of miRCURY LNA microRNA Arrays, we tested the serumcollected before and after radiotherapy，and screened differentially expressed miRNAs inserum. Then qRT-PCR (Taqman probe method) was used to verify, meanwhile，detecting the serum Cyfra21-1, Ki-67,SCCA, LDH, ALP levels after radiotherapy, and analysised therelationship between these biomarkers and the recurrence and metastasis of nasopharyngealcarcinoma,searching for the possible tumor marker to evaluate nasopharyngeal carcinomasurvival time.Methods:1.We enrolled3cases of recurrence or distant metastasis patients in5years and3cases of clinical remission patients in5years,collecting the paired serum before and aftertreatment.With the application of miRCURY LNA microRNA Arrays, screening thedifferent expression spectrum of serum miRNAs.2.Collected29cases of recurrence or distant metastasis patients in5years and19cases of clinical remission patients in5years,collecting the paired serum before and aftertreatment.With the application of qRT-PCR (Taqman probe method),verifying the differentexpression spectrum.3.Collected the NPC paraffin blocks and serum after treatment of patients above,immunohistochemistry was used to detect the expression of Ki-67, chemiluminescencemethod was used to detect the expression of serum Cyfra21-1, SCCA,and continuousmonitoring assay was used to detect the expression of serum LDH, ALP.4.Analysising the relationship between these biomarkers and the survival time.Results：1.The miRNAs chip showed that serum miRPlusE1253is up-regulated in recurrence ordistant metastasis group than in clinical remission group before treatment, while serummiR-22, miR-29a, miR-720, miR-665, miR-26b, miR-125b is down-regulated in recurrenceor distant metastasis group than in clinical remission group before treatment,serummiRPlusE1072is up-regulated in recurrence or distant metastasis group than in clinicalremission group after treatment, serum miR-22, miR-29b, miR-143, miR491-3p, Let-7c,miR-720, miR-550, miR-30c is down-regulated in recurrence or distant metastasis groupthan in clinical remission group after treatment.All the difference were more than2times.2.qRT-PCR showed the expression level of miR-26b, miR-29a, miR-29b, miR-143,miR-125b was no significant difference between the recurrence or distant metastasis groupand clinical remission group.3.The combined detection of serum miR-29b, miR-125b before treatment and Ki-67in patients with nasopharyngeal carcinoma can established a equation to predict recurrence ordistant metastasis of nasopharyngeal carcinoma，specificity and sensitivity were75.9%and78.9%, positive predictive value and negative predictive values were84.6%and68.1%.4.COX regression analysis showed that serum miR-29a and miR-125b beforetreatment were independent prognostic factors for progression free survival and overallsurvival.Conclusion：1.The serum miR-29a and miR-125b before treatment are independent prognosticfactors for progression free survival and overall survival. Combined detection of serummiR-29a and miR-125b can be used to evaluate the progression free survival and overallsurvival of the patients.2.Combined detection of serum miR-29b, miR-125b before treatment and Ki-67canbe used to predict recurrence or distant metastasis of nasopharyngeal carcinoma patients. |
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