The Value Of Ultrasonic Parameters Am/At Combined with CYP27A1 Gene in the Diagnosis of Pediatric Cervical Lymphoma and the Functional Study of CYP27A1 Gene
Objective To investigate the value of ultrasound parameter ratio of medulla department and total area of lymph nodes(Am/At)combined with cytochrome P450 family 27 subfamily A member 1(CYP27A1)gene in the diagnosis of pediatric cervical lymphoma and the function of CYP27A1 gene.Methods The Am/At of children with benign and malignant cervical lymph node lesions were examined by ultrasonography.The mRNA level of CYP27A1 in peripheral blood lymphocytes was detected by RT-qPCR.The benign and malignant cervical lymphadenopathy were diagnosed by ROC curve analysis of Am/At and CYP27A1.OCI-LY8 cells were divided into 5 groups:Control group,shNC group,shCYP27A1 group,OENC group and OECYP27A1 group.OCI-LY8 cells in Control group were not transfected,while Lipofectamine 2000 was used to transfect shRNA or lentivirus into OCI-LY8 cells.MTT method was used to detect cell proliferation,flow cytometry was used to detect apoptosis,and Transwell was used to detect cell invasion.The mRNA level of CYP27A1 in OCI-LY8 cells was detected by RT-qPCR.The protein levels of CYP27A1,Bcl-2,Bax,MMP-2,MMP-9,p-PI3K,PI3K,p-AKT and AKT in OCI-LY8 cells were detected by Western blot.Results Compared with the benign group,the Am/At of malignant group significantly decreased(0.31±0.13 vs 0.15±0.08,t=6.090,P＜0.001),the level of CYP27A1 mRNA in the peripheral blood lymphocytes of the malignant group was significantly increased(0.33±0.14 vs 0.65±0.22,t=8.120,P＜0.001).Taking＞0.34 as the cutoff value,the AUC of Am/At combined with CYP27A1 in the diagnosis of benign and malignant cervical lymph nodes was 0.937(Z=18.324,P＜0.0001),the sensitivity was 90.32％,and the specificity was 82.35％.Compared with control group and shNC group,the levels of CYP27A1 mRNA and protein in shCYP27A1 group were decreased(P＜0.05),the protein levels of Bcl-2,MMP-2 and MMP-9 were decreased(P＜0.05),the phosphorylation levels of PI3K and AKT were decreased(P＜0.05),the relative cell viability and number of invasion cells were decreased(P＜0.05),the protein levels of Bax and apoptosis rate were increased(P＜0.05).Compared with control group and OENC group,the levels of CYP27A1 mRNA and protein in shCYP27A1 group were increased(P＜0.05),the protein levels of Bcl-2,MMP-2 and MMP-9 were increased(P＜0.05),the phosphorylation levels of PI3K and AKT were increased(P＜0.05),the relative cell viability and number of invasion cells were increased(P＜0.05),the protein levels of Bax and apoptosis rate were decreased(P＜0.05).Conclusion This study shows that Am/At combined with CYP27A1 detection may be helpful to improve the diagnostic accuracy of children with lymphoma,and down-regulation of CYP27A1 may inhibit the proliferation and invasion of DLBCL cells by inhibiting PI3K/AKT signal pathway.
Pediatric Cervical LymphomaUltrasonic ParametersCYP27A1Lymph NodesRatio of Medulla Department and Total Area of Lymph Nodes
万方数据
维普
