The Mechanism Of Action Of UVRAG Gene In Malignant Transformation Of MDS And The Effect Of Relevant Gene Mutations On The Prognosis Of MDS Transplantation

Part 1:Study on the mechanism of action of UVRAG gene in MDS malignant transformationObjective:Myelodysplastic syndromes(MDS)are heterogeneous clonal diseases that originate from bone marrow hematopoietic stem cells.Such patients present with abnormal morphology of bone marrow cells,a decrease in primary or multilineage cells,and a high risk of developing acute myeloid leukemia.The autophagy related gene UVRAG is involved in the autophagy process of cells.Our previous study found that the expression of UVRAG gradually decreased as MDS turned from low-risk to high-risk,which was negatively correlated with the malignant progression of MDS.This experiment intends to study the mechanism of the action of UVRAG in the malignant transformation of MDS.Methods:1.Transfection of MDS into leukemia cell line SKM-1 with targeted UVRAG specific interfering RNA(siRNA-UVRAG).2.qRT-PCR was used to detect the expression levels of UVRAG,Bcl-2,LC3 and Beclin-1 before and after siRNA-UVRAG interference.3.CCK-8 method was used to detect the proliferation of SKM-1 cells before and after siRNA-UVRAG interference.4.Annexin v-FITC/PI double staining and flow cytometry was used to detect the apoptosis of SKM-1 cells.Results:With the down-regulation of UVRAG,the expressions of LC3 and Beclin-1,markers of autophagy,also showed a downregulation trend and autophagy decreased.The cell proliferation rate of siRNA-UVRAG interference group was higher than that of control group.The apoptosis rate in the siRNA-UVRAG interference group was lower than that in the control group.Conclusion:1.UVRAG plays a positive regulatory role on autophagy,and is coordinated with LC3 and Beclin-1.2.In MDS malignant transformation,UVRAG may play a role by promoting autophagy and apoptosis.Part2:Effect of MDS-related gene mutations on the prognosis of transplantationObjective:In recent years,more and more studies have shown that MDS related gene mutations are significantly correlated with the prognosis of transplantation.This study analyzed and statistically analyzed the common gene mutations in patients with myelodysplastic syndrome(MDS),and discussed its influence on the total survival after hematopoietic stem cell transplantation in MDS patients.Methods:Sixty-eight patients diagnosed with MDS and transplanted in the First Affiliated Hospital of Suzhou University from October 2013 to August 2018 were selected.Genetic mutations in bone marrow samples diagnosed before transplantation were detected by second-generation gene sequencing.Statistical methods were used to analyze the effects of different genetic mutations on the prognosis of transplantation.Results:The mutations of NRAS,TP53 and GATA2 were related to the overall survival period of OS after hematopoietic stem cell transplantation.The overall survival time of patients with these gene mutations was shorter than that of patients with the wild-type gene,and the difference was statistically significant.Conclusion:1.Mutations in the genes of NRAS,TP53 and GATA2 lead to shortened overall survival and poor prognosis after transplantation.2.Mutations of NRAS,TP53 and GATA2 genes are independent prognostic factors of MDS patients after transplantation,and whether there are these mutations can be used as an important indicator to evaluate the prognosis of MDS patients after transplantation.