EXTRACTION OF FACTORS RELATED TO THE ONCOGENESIS, PROGRESSION AND SENSITIVITY TO THE THERAPEUTIC AGENTS IN HUMAN MYELOMA CELLS AS IN VITRO MODEL

我们分析了ATRA（全反式维甲酸）、双膦酸盐（YM529）、抗her2单克隆抗体、HMG-CoA还原酶抑制剂（他汀类药物，如普伐他汀和辛伐他汀）、缺氧和沙利度胺对人骨髓瘤细胞的影响。12个骨髓瘤细胞系中有2个在补充ATRA后表现出生长增强，其特征是IL-10的产生、膜Fas的下调和p21/Cip1 CDK-I信息上调的降低。YM529对人骨髓瘤细胞株的生长抑制作用呈剂量依赖性。细胞聚集在细胞周期的[2n< <4n]，随后形成凋亡的亚g1部分。ATRA、沙利度胺或α干扰素联合治疗可增强YM529的生长抑制作用。虽然通过RT-PCR分析的HER家族基因mRNA水平在骨髓瘤细胞中明显低于乳腺癌细胞，但一些细胞系表达了一定数量的HER2和HER4蛋白。此外，抗her2单克隆抗体rhumAbHER2在研究的8个骨髓瘤细胞系中有6个产生了显著的生长抑制作用，这些抑制作用与先前研究的乳腺癌细胞相似。rhumAbHER2诱导p21家族CDK-Is上调，VEGF基因下调。此外，联合抗雌激素治疗具有累加性生长抑制作用。我们证实与非骨髓瘤血液系相比，骨髓瘤中VEGF的表达和产生水平增加，骨髓瘤中缺氧抵抗和增强VEGF- a的产生，以及由于沙利度胺诱导的tnf - α上调引起的凋亡和G1停滞而直接抑制骨髓瘤细胞的生长。尽管浓度高于临床使用的浓度，10个骨髓瘤系中有4个被普伐他汀抑制生长。相关因素的研究表明IL-6的抑制作用是重要的。事实上，在不表达IL-6的KMS-21BM细胞中，rhIL-6消除了普伐他汀诱导的生长抑制。当用不同浓度的辛伐他汀以剂量依赖的方式培养时，大多数骨髓瘤细胞系（13株中的12株）显示出生长抑制。辛伐他汀联合其他生物反应调节剂如ATRA或DEX对生长有额外的影响。此外，抗氧化物可阻止辛伐他汀诱导的生长抑制和细胞凋亡。少

We have analyzed the effects of ATRA(all-trans retinoic acid), bisphosphonate(YM529), anti-HER2-monoclonal antibody, HMG-CoA reductase inhibitors(STATINs, e.g., pravastatin, and simvastatin), hypoxia and thalidomide on human myeloma cells. Two out of 12 myeloma cell lines showed enhanced growth on supplementation of ATRA and were characterized by IL-10 production, down regulation of membrane Fas and reduced upregulation of p21/Cip1 CDK-I message. The growth inhibitory effect of YM529 on human myeloma cell lines was dose dependent. The cells accumulated in [2n< <4n] of the cell cycle and subsequently formed an apoptotic sub-G1 fraction. Combined treatment with ATRA, thalidomide, or interferon alpha enhanced the growth inhibitory effects of YM529. Although the mRNA levels of HER family genes analyzed by RT-PCR were significantly mower in myeloma cells than breast cancer cells, some cell lines expressed a certain amount of HER2 and HER4 proteins. In addition, an anti-HER2 monoclonal antib … More ody, rhumAbHER2, caused significant growth inhibition in six out of eight myeloma cell lines studied and these inhibitory effects were similar to those in the breast cancer cells studied previously. The rhumAbHER2 induced upregulation of p21 family CDK-Is and down-regulation of VEGF genes. Moreover, combination treatment with antiestrogen had an additive growth inhibitory effect. We demonstrated increased expression and production levels of VEGF in myeloma compared to non-myelomatous hematological lines, resistance to hypoxia and enhancement of VEGF-A production by hypoxia in myeloma, and direct growth inhibition of myeloma cells due to apoptosis and G1 arrest caused by TNF-alpha upregulation induced by thalidomide. Although the concentrations were higher than those used clinically, 4 out of 10 myeloma lines showed growth inhibition by pravastatin. The study of factors related to the inhibition indicated IL-6 is important. Indeed, rhIL-6 abolished pravastatin-induced growth inhibition in KMS-21BM cells which did not express IL-6. Most of myeloma lines(12 out of 13) examined showed growth inhibition when cultured with various concentrations of simvastatin in a dose-dependent manner. Simvastatin in combination with other biological response modifiers such as ATRA or DEX had additional effects on growth. In addition, anti-oxides prevented the simvastatin-induced growth inhibition and apoptosis. Less