| Expression of epoxyeicosatrienoic acids in multipleAim:To investigate the levels of 11,12-EET and 14,15-EET in multiple myeloma cells and serum of multiple myeloma patients.Methods:Elisa assay was used to examine 11,12-EET and 14,15-EET levels in U266 and RPMI8226 cells. Serum of multiple myeloma patients was isolated, and EETs, VEGF levels in the serum were also detected by Elisa assay. The CYP2J2 mRNA levels in MM cells were examined by RT-PCR.Results:The result of Elisa assay showed that multiple myeloma cells contained 11,12-EET and 14,15-EET.2×106 U266 cells contained 239789.80±50297.81pg/ml of 11,12-EET and 642293.81±28930.74pg/ml of 14,15-EET.11,12-EET was 188487.56±58858.03pg/ml and 14,15-EET was 536489.31±21331.71pg/ml in the same number of RPMI8226 cells. Serum of multiple myeloma patient also contained 11,12-EET and 14,15-EET, which were significantly higher than healthy control (P<0.01). But in our results, the EETs concentrations were not correlated with VEGF and the poor prognostic factors of MM such as LDH andβ2-microglobulin (P>0.05). The subfamily member of P450 expoxynases CYP2J2 mRNA was detected in U266 cells, but was not detected in RPMI8226 cells.Conclusion:Both 11,12-EET and 14,15-EET are expressed in multiple myeloma cells and serum of multiple myeloma patients. The EETs levels in patients serum are higher than healthy control, which suggest EETs may play an important role in multiple myeloma. Aim:In the present study, we investigate the effect of P450-EETs pathway in multiple myeloma proliferation and the related mechanism.Methods:Elisa assay was used to examine 11,12-EET and 14,15-EET levels in U266 and RPMI8226 cells treated by 17-ODYA. MTT assay was used to investigate the effects of 11,12-EET,14,15-EET and 17-ODYA on U266 and RPMI8226 proliferation. Flow cytometry was used to examine the apoptosis and cell cycle changes in multiple myeloma cells treated by 17-ODYA. The expression of apoptosis and cell cycle related protein was detected by western blotting.Results:17-ODYA decreased 11,12-EET and 14,15-EET levels in U266 and RPMI8226 cells. Addition of exogenous EETs promoted the proliferation of MM cells, whereas 17-ODYA inhibited the viability and exogenous EETs reversed 17-ODYA-mediated decrease of proliferation.17-ODYA enhanced MM cells apoptosis and induced cells arrested at G0/G1 phase through regulating the expression levels of related proteins to suppress the proliferation of MM cells.Conclusion:P450-EETs pathway plays an important role in multiple myeloma proliferation, and 17-ODYA surpresses this effect and may be a new tool for multiple myeloma treatment. Aim:The goal of the present study is to investigate the expression of epoxyeicosatrienoic acids (EETs) and its role in the angiogenesis of multiple myeloma (MM).Methods:MM cell lines of U266 and RPMI8226 were cultured, and the EETs levels (11,12-EET and 14,15-EET) in the supernatant were determined by Elisa assay. Human umbilical vein endothelial cells (HUVEC) were cultured and used for analysis of the angiogenesis activity of the two MM cell lines, which was examined in vitro and in vivo with the usement of MTT, chemotaxis, tube formation and matrigel plug assays.Results:We confirmed that 11,12-EET and 14,15-EET were secreted into the supernatant of the culutured MM cells. The levels of the two EETs in the supernatant of U266 cells were significantly higher than those in the RPMI8226 cell supertanant (P<0.05), which was positively correlated with the respective angiogenesis activity of the two different MM cell lines. 17-octadecynoic acid (17-ODYA), as a specific inhibitor of P450 enzyme, suppressed HUVEC proliferation and tube formation induced by multiple myeloma cells. Furthermore,17-ODYA decreased the EETs levels in the supernatant of multiple myeloma.Conclusion:These results reveal that EETs may play an important role in the angiogenesis of multiple myeloma, and the inhibitor 17-ODYA suppresses this process.Aim:To investigate the effects of 17-ODYA on the motility of multiple myeloma cells and the mechanisms involved.Methods:The migration and invasion assay were performed by transwell model analyses. The activity of matrix metalloproteinase (MMP)-2 and-9 were detected by gelatin zymography analysis. The expression of MMP-2, MMP-9, p-Akt, Akt and HIF-1αwere determined by Western blotting.Results:17-ODYA inhibited the motility and EETs levels of myeloma cells in a time-dependent manner. Incubated of myeloma cells with 17-ODYA resulted in the reduction of MMP-2 and MMP-9 protein expression and gelatinolytic activity.17-ODYA down-regulated the phosphorylation of Akt, but had no effect on HIF-1αConclusion:17-ODYA inhibits multiple myeloma cells migration and invasion. The inhibition of invasion is associated with the reduction of the enzymatic activity and expression of MMPs, which may be regulated by the phosphorylation of Akt. |
PDF Full Text Request