Dysregulation Of MiR-320 Family Members As Potential Diagnostic And Prognostic Biomarkers In Myelodysplastic Syndrome

1.ObjectiveTo explore the expression of miR-320 family members in myelodysplastic syndromes and analyze the the correlation between miR-320 family expression level and clinical parameters of MDS patients.The diagnostic role of miR-320 family members in MDS was analysed by ROC curve.The prognostic role of miR-320 family members in MDS was further explored by survival analysis,ensuring the miR-320 family members could be used for the diagnosis and prognosis of MDS.2.MethodsFirst of all,we analyzed the data screened by microRNA microarray technology,and found that there were significant differentially expressed microRNAs in MDS patients ——miR-320 family.Then we expanded the sample size to 82 patients with MDS and 24 healthy donors.We used q RT-PCR technology to detect the expression of miR-320 family members in bone marrow of MDS patients and healthy donors to verify the microarray results,and collected the clinical parameters of MDS patients(age,gender,peripheral blood,etc.)and follow up MDS patients to record the survival status of MDS patients,and the correlation between the expression of miR-320 family members and clinical parameters of MDS patients was analyzed.ROC curve was drawn to analyze the diagnostic value of miR-320 family members to MDS.Kaplan-Meier methods was used to analyze the relationship between the expression of miR-320 family members and the median survival time of MDS patients.3.Results(1)Based on the microarray data,we found that all members of the miR-320 family,including miR-320 a,miR-320 b,miR-320 c,miR-320 d and miR-320 e,were all highly expressed in bone marrow of MDS patients.By further expanding the sample size and typing according to IPSS,the samples were divided into three groups: relatively low-risk group(low risk +intermediate risk I),relatively high-risk group(intermediate risk II + high risk),s AML group.q RT-PCR was used to detect the expression of miR-320 family in each subgroup.The results showed that except for the expression of miR-320 d in the relatively low-risk group which did not reach the statistical significance(P= 0.0537),all miR-320 family members were significantly increased in each subgroup of MDS.The expression of miR-320 family members in the relative high risk group and the s AML group was significantly higher than that in the low risk group(P < 0.05).(2)We divided MDS patients into miR-320 high expression group and miR-320 low expression group according to the median expression of each member of mir-320 family.The clinicopathological parameters of the two groups were compared and found that: the count of bone marrow blasts,the ratio of advanced WHO type,the rate of abnormal chromosome of miR-320 c high expression group and miR-320 d high expression group were significantly higher than that of miR-320 c low expression group and miR-320 d low expression group(P = 0.003,P = 0.047;P = 0.006,P = 0.016;P < 0.001,P = 0.024);the ratio of advanced IPSS risk stratification of high expression group of all members of miR-320 family was higher than that of low expression group(P=0.001,P=0.024,P<0.001,P<0.001,P=0.012).(3)ROC curve analysis was used to explore the diagnostic role of miR-320 family members in MDS.The results showed that all members of miR-320 family had diagnostic value for MDS.The area under the curve was as follows:miR-320 a,0.9037(95% CI: 0.8297-0.9777,P < 0.0001);miR-320 b,0.7515(95%CI: 0.6496-0.8535,P = 0.0002);miR-320 c,0.9647(95% CI: 0.9345-0.9949,P <0.0001);miR-320 d,0.8064(95% CI: 0.7075-0.9053,P<0.0001);miR-320 e,0.9019(95% CI: 0.8407-0.9632,P<0.0001).(4)We used Kaplan-Meier methods to compare the median survival time of miR-320 high expression group and miR-320 low expression group.It was found that the median survival time of miR-320 c and miR-320 d high expression group was significantly lower than that of miR-320 c and miR-320 d low expression group(P=0.002,P=0.032).4.Conclusions(1)The expression of miR-320 family was significantly up-regulated in the bone marrow of MDS patients,and the high expression of miR-320 c and miR-320 d was significantly correlated with high BM blasts,advanced WHO type,and poor karyotype of MDS patients.High expression of all members of miR-320 family was correlated with advanced IPSS type of MDS patients.(2)miR-320 family has diagnostic value for MDS.High expression of miR-320 c and miR-320 d is associated with poor prognosis of MDS.