Effects of HDAC Inhibitor Scriptaid on IM9 Cell Line and Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effect of HDAC inhibitor Scriptaid on multiple myeloma IM9 cells and preliminarily clarify the mechanism of Scriptaid-induced cell apoptosis.</p><p><b>METHODS</b>The cell viability, cell cycle and cell apoptosis were measured by CCK8 assay and flow cytometry respectively, the relative target gene expression levels were detected by RT-PCR, the effect of Scriptaid on p21 promoter activity was detected by using luciferase reporter assay.</p><p><b>RESULTS</b>Scriptaid inhibited IM9 cell viability in a dose-dependent manner. Scriptaid induced IM9 cell cycle arrest at G/M phase in a dose-dependent manner. Scriptaid triggered IM9 cell apoptosis was obviously, the mRNA levels of apoptosis-related proteins Caspase 9, Caspase 3 and PARP1 were also activated. The apoptosis-associated factors BAD, PTEN and p21 increased following treatment with different dose of Scriptaid, meanwhile, p21 promoter activity was also activated significantly.</p><p><b>CONCLUSION</b>HDAC inhibitor Scriptaid can promote IM9 cell apoptosis by transcriptional activation of p21 promoter in concentration-dependent manner.</p>

Effect of Interfering the ADAM10 by Lentivirul Vector-Mediated shRNA on Multiple Myeloma MM.1S Cell Proliferation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of interfering ADAM10 on proliferation and apoptosis of multiple myeloma MM.1S cells, and its possible mechanism.</p><p><b>METHODS</b>Four pairs of shRNA-coding sequences directed against different sites of ADAM10 mRNA were designed and inserted into lentiviral vector plasimd pLVshRNA-EGFP(2A) Puro for constructing the sh/ADAM10-1, sh/ADAM10-2, sh/ADAM10-3, sh/ADAM10-4 and sh/Con. These plasmids and lentiviral packaging plasmids were co-transfected into the packaging cells 293FT, then the virus particles were collected and the viral titer was assayed after concentration, and these viral particles were transfected to MM.1S cells. The flow cytometry was used to sort GFPcells. Real-time quantitative PCR, and Western blot were used to detect the effect of interfering the ADAM10 gene by lentiviral vector mediated shRNA. The proliferation-inhibition curve was plotted by CCK-8 method, the cell viability and apoptosis were detected by flow cytometry with Annexin V and 7-AAD staining, the transcripts of pro-apoptosis gene BAD, BAK, BIK, anti-apoptotic genes BCL-2, c-Myc and Notch1 target gene Hes-1 were detected by real-time PCR.</p><p><b>RESULTS</b>Lentivirus vector was successfully constructed, that could specifically interfere ADAM10 expression. Interfering ADAM10 gene could inhibit the MM.1S cell proliferation and induce apoptosis. After the interferencing ADAM10 gene the mRNA levels of pro-apoptosis gene BAD, BAK and BIK were increased, and the mRNA levels of anti-apoptotic genes BCL-2 and c-Myc were reduced. Q-PCR results showed that the mRNA level of Notch1 were increased, but that of Hes-1 were reduced.</p><p><b>CONCLUSION</b>Down-regulated ADAM10 expression can significantly inhibit multiple myeloma MM.1S cell proliferation and promote the apotosis. Its mechanism may be related to Notch1 signaling pathways.</p>