Detection Of The Expression Level Of MicroRNAs In Serum Of Osteosarcoma Patients Using Real-time Fluorescent Quantitative PCR

OBJECTIVE:To detect the expression lever of relevant miRNAs in serum of osteosarcoma patients and find out the potential candidate miRNA for diagnosis and treatment of osteosacrcoma.MATERIALS AND METHODS:The expression lever of relevant miRNAs in serum was detected using real-time fluorescent quantitative PCR(stem-loop RT-qPCR, TaqMan probe). Serum specimens were divided into three groups:1,the group of osteosarcoma patients who had not chemotherapy; 2,the group of osteosarcoma patients who had chemotherapy; 3,healthy control group. Each group had 10 serum specimens. Detect the expression lever of mir-29c, mir-132, mir-199a-3p, mir-221 in serum. Choose U6 snRNA as endogenous control. The correlation between the expression lever of each microRNA. and its clinicopathologic features was analyzed by SPSS 17.0 statistical software.α=0.05 was regarded as the statistical criterion.RESUITS:1. Four miRNAs can be detected in all serum specimens.2. The high expression level of miR-29c and miR-221 were detected in serum of osteosarcoma patients, compared with healthy control group, there is statistics difference(P<0.05), while miR-199a-3p and miR-132 had no significant difference(P>0.05). 3. There is no correlation between the expression level of four miRNAs in serum of osteosarcoma patients and histologic grade,histologic subtype,Enneking surgical staging,ALP(P>0.05).4. The expression lever of each miRNA has no significant difference between the group of osteosarcoma patients who had not chemotherapy and the group of osteosarcoma patients who had chemotherapy(P>0.05).CONCIUSION:1. There are miRNAs in human serum, which can be detected conveniently by real-time fluorescent quantitative PCR method. Stem-loop RT-qPCR(TaqMan probe) is a sensitive, specific and convenient method to detect miRNA quantitatively.2. miR-29c and miR-221, which have high expression level in the serum of osteosarcoma patients, can be the candidate of serum biomarker of osteosarcoma.3. There may be some specific serum-miRNA profile of osteosarcoma patients.