双靶点药物NL-101的抗多发性骨髓瘤活性及其机制研究

Antitumor effects and the mechanisms of dual-targeting drug NL-101 on human multiple myeloma

研究具有组蛋白去乙酰化酶(histone deacetylase,HDAC)抑制和DNA损伤双靶点化合物NL-101的体内外抗多发性骨髓瘤活性及其机制研究。在10株多发性骨髓瘤细胞株中,采用CTG法检测化合物NL-101对肿瘤细胞增殖的影响以及在RPMI 8226细胞中NL-101与硼替佐米的联用效果,采用流式细胞仪检测NL-101对细胞周期的影响,Western blot检测NL-101对乙酰化组蛋白H3、乙酰化α-Tubulin和磷酸化Histone H2A.X的表达影响,并在RPMI 8226多发性骨髓瘤细胞皮下移植瘤模型中考察NL-101单用以及与硼替佐米联用的抑瘤效果。研究结果表明,NL-101能同时产生HDAC抑制剂和DNA烷化剂双重活性,不但抑制肿瘤细胞组蛋白去乙酰化水平,而且增加DNA损伤,最终诱导细胞周期阻滞和细胞凋亡。在细胞水平和动物移植瘤模型中均表现出较强的抗肿瘤活性,优于阳性对照药美法仑,而且与硼替佐米联用具有协同增效作用。NL-101有望成为一个全新的抗多发性骨髓瘤药物。

The antitumor activities of NL-101, aHDACi/DNA damage dual-targeting drug, on human multiple myeloma in vitro and in vivo were studied. Furthermore, the primary mechanisms were revealed. We detected the anti-proliferative activity of NL-101 on 10 human multiple myeloma cell lines, and the combinational effect of NL- 101 and bortezomib on RPMI 8226 cell line. The inducing effects of NL-101 on cell cycle arrest and apoptosis were detected by FACS. The effects of NL-101 on acetyled-Histone H3, total Histone H3, acetyled a-Tubulin, total a-Tubulin, phospho-Histone H2A. X and total Histone H2A. X were evaluated by Western blott. We also demonstrated the antitumor activity of NL-101 and the combinational effect of NL-101 and bortezomib on RPMI 8226 xenograft tumor model in vivo. Results showed that NL-101 possessed strong antitumor activities on human multiple myeloma cells in vitro and in vivo. NL-101 exhibited significant HDAC inhibitory activity and DNA alky-lating activity. NL-101 not only inhibited histone deacetylation level, but also increased the DNA damage in multi-ple myeloma cells. Meanwhile, NL-101 induced cell cycle arrest and apoptosis. Also, the synergistic effect of NL- 101 was discovered when combined with bortezomib in vitro and in vivo. These data demonstrated that NL-101 may be a potent agent for the treatment of human multiple myeloma in future.