Solanum Nigrum Total Alkaloids Induced Apoptosis Of Myeloma Cells And Notch1Effects

Myeloma RPMI8226cells in this topic selected for the study, the total alkaloids of Solanum nigrum test object, as the main role of aspects of RPMI8226cells within the Notchl, of Annexin V-F1TC/PI flow cytometry was used to observe the myeloma RPMI8226cells withereddeath, Notch1mRNA of expression detected by RT-PCR. To investigate the apoptotic effect of Solanum nigrum alkaloids myeloma RPMI8226cells, and Notch1.Objective:1.observation of Solanum nigrum alkaloids on the role of apoptosis in myeloma cell lines RPM18226cells, to find the optimal concentration and the optimum time of Soianum nigrum of total alkaloids.Notch1signaling pathway2.The total alkaloids from the role of Solanum nigrum in RPM18226myeloma cells, try to elaborate the mechanism of inhibition of cell proliferation and induce apoptosis.Methods:1.RPM18226cells cultured and passaged every2to3days once the experiment select logarithmic phase cells.2.By MTT method to explore the Solanum nigrum total alkaloids inhibit RPMI8226cell growth activity, concentration and time to join the Solanum nigrum total alkaloids,set800,400.200,100.50,25.12.5.6.25.3.125mg/L. a total of nine concentrations ofgradient,set up another blank control group, n=5wells. Continue to foster.24,48and72hours, cell growth activity by MTT assay.3.Annexin V/PI flow cytometry assay of apoptosis cells in each group dosing after48h, centrifuged cells were collected, washed two times the pre-cooling PBS at4℃, and then use250ul binding buffer to resuspend the cells, and adjust its concentration to106/ml, and then take100ul cells were suspended in5ml flow tubes, add5ul by Annexin V-FITC and10ul20ug/ml propidium iodide solution, mix well after dark at room temperature for15minutes, then resuspended in400ulPBS,400mesh filter, the sample cell apoptosis was detected by flow cytometry (FACS). For apoptosis data ModFitLT software apoptosis relative quantitative analysis.4.RT-PCR assay of Notch1gene expression were taken after total alkaloids from various Solanum nigrum intervention cells, extracted phase cells mRNA by reverse transcription technology, synthetic phase cell cDNA by the design of the Notchlgene primers, PCR amplification of Notch1gene sequence and the gene of the beta-action internal control, the use of the intervention in the software to detect cell expression of Notch1gene.5.The statistical analysisSPSS17.0software analysis, data were expressed as mean±standard deviation related to sample non-parametric analysis of variance, P<0.05significant difference, P>0.05for the difference was i(?)ficant resistance.Results:1.Solanum nigrum total alkaloids inhibit RPMI8226cell proliferation, was dose and time dependent. RPMI8226cells48h after the25mg/L of Solanum nigrum total alkali treatment in RPMI8226cell growth is inhibited. 2.Annexin Ⅴ/PI method confirmed the total alkaloids of Solanum nigum can be induced by human multiple myeloma cell lines RPMI8226apoptosis.3.RT-PCR results showed that Solanum nigrum total alkaloids can down-regulate the expression of Notch1and the role of concentration and time of a certain concentration of total alkaloids of Solanum nigrum.Conclusion:1.Solanum nigrum total alkaloids inhibit myeloma cell lines RPM18226cell proliferation was dose and time dependent.2.Annexin Ⅴ/PI method confirmed the total alkaloids of Solanum nigrum can be induced by human multiple myeloma cell lines RPMI8226apoptosis.3.Solanum nigrum alkaloids inhibit Notch1expression in myeloma cell lines RPMI8226cells. The mechanism of the Solanum nigrum total alkaloids induced apoptosis of myeloma cells may be caused by down-regulating Notch1downstream of NF-κB inactivation resulting in conduction was inhibited, thus affecting the expression of other apoptosis-related factors or related pathways conduction interruption.