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[Objective] To study natural compounds isothiocyanate, benzyl ester (PHI) in vitro to inhibit the proliferation of multiple myeloma cell line U266, the analysis of the impact of PHI myeloma cell methylation transferase (DNMT), explore the PHI inhibition mechanism the myeloma cells 増 colonization, as a new treatment for multiple myeloma drugs to provide experimental evidence for the PHI. [Methods] By depicting cell growth curve, MTT method observed PHI U266 cell growth inhibition; PHI induced apoptosis of cells was observed by DAPI staining; U266 cell cycle by flow cytometry analysis of PHI, RT -PCR analysis of DNA methyltransferase (DNMT) 1,3 a, 3b in multiple myeloma cell line U266 and normal human peripheral blood mononuclear cells expressed; analyzed by RT-PCR technology the different concentration PHI role U266 cells after DNMT1 DNMTs activity in cells, 3a, 3b in the cells and using the class detected by ELISA with different concentrations PHI role; using the methylation status of methylation PCR analysis of P16 gene in U266 cells, and observe PHI U266 cell line P16 gene methylation status. 【Results】 1, RT-PCR results showed DNMT1, 3a, 3b myeloma cell line U266 high expression and low expression in normal human cells. Use 1.25umol / L of 2.5umol / L of 5umol / L 10umol/LPHI U266 cells after treatment, and the proliferation ability was significantly suppressed 48 hours inhibition rates were 0.284 ± 0.089,0.554 ± 0.079,0.753 ± 0.053,0.936 ± 0.067 (p lt; 0.05), MTT resulting IC50 2.48.2 DAPI staining showed that apoptosis. 3, cell cycle analysis, PHI inhibited myeloma cells growth with colonization, the growth of tumor cells in G1 phase is discontinued. 4, PCR analysis of tumor cells DNMTs expression DNMTmRNA express abnormally high levels, RT-PCR: different concentrations of PHI cells 48 hours after treatment, with the blank control than, RT-PCR display U266 cells expressions of DNMT1, DNMT3a, and DNMT3b mRNA of decreased expression (p lt; 0.05) and dose-dependent manner. 5, using ELISA techniques in detecting tumor cells DNMT activity found PHI treatment DNMT activity was significantly inhibited. 6, methylation PCR technique detected the P16 gene hypermethylation in U266 cells, the use of PHI processing allows the reversal of the P16 gene hypermethylation status. [Conclusion] U266 cells DNMT expression was significantly higher, significantly different from normal cells. 2, PHI treatment myeloma cell line U266, significant inhibition of tumor cell DNMT activity, induction of apoptosis, DNMTmRNA reduced expression in a dose-dependent manner. 3, PHI reversal of the P16 gene is hypermethylated in U266 cells. 4, DNMT available as a new target for the treatment of myeloma, PHI is a new formulation of the potential treatment of myeloma.
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