ALBERT

Description: Tumor relapse and cytomegalovirus(CMV) infection are very important concerns in the therapy of hematopoietic malignancies by bone marrow transplantation.It was reported that there was a higher rate of CMV reactivation in patients with multiple myeloma(MM) after autologous or CD34+ stem cell selected transplantation,but no attention so far has been given to a possible pathogenetic interplay between CMV and MM cells.CMV could infect many kinds of cells, and could inhibit apoptotic responses in several cell systems.Our studies were to investigate the alteration of apoptosis in MM cell line cells infected by CMV as well as the possible mechanism. After CMV AD169 was propagated in HF fibroblasts,MM cell line KM3 and RPMI 8226 cells were infected by 100,10,1 TCID50 of CMV,then were cultured with serum-free RPMI 1640.RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene,Flow Cytometry was used to detect the CMV pp65 antigen positive cells and transmission electron microscope was used to detect CMV particles in the cells as well as the morpholoy and inner structure of the apoptotic MM cells. AnnexinV-FITC and Propidium Iodide Staining Solution were added to MM cells and apoptotic cells were detected with Flow Cytometry.To explore the possible mechanism that CMV infection inhibit the apoptotic responses in MM cells,RT-PCR assay was used to detecte the mRNA expression of IL-6 and VEGF in CMV-infected and mock-infected cells; ELISA assay was used to detecte the protein expression of IL-6 in the culture medium; EMSA assay was used to detecte the NF-κB activity of the cells.We got these results as below: CMV-infected MM cells could express IE mRNA compared with the uninfected;CMV particles could be found in the cells infected by 100 TCID50 of CMV compared with the mock-infected cells,and there were much more CMV pp65 antigen positive cells when KM3 cells were infected by 100,10 TCID50 of CMV,which was significant higher than the control group P