van de Donk N W C J, Kamphuis M M J, van Dijk M, Borst H P E, Bloem A C, Lokhorst H M
Department of Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
Leukemia. 2003 Jan;17(1):211-9. doi: 10.1038/sj.leu.2402768.
An antisense oligodeoxynucleotide (ODN) complementary to the first six codons of the Bcl-2 mRNA, G3139 (oblimersen sodium; Genasense), has been shown to downregulate Bcl-2 and produce responses in a variety of malignancies including drug-resistant lymphoma. Incubation of ex vivo purified plasma cells from patients with multiple myeloma (MM) with carboxyfluorescein (FAM)-labeled antisense ODNs resulted in a time- and dose-dependent uptake in the cytoplasm and nucleus. No major differences in uptake of Bcl-2 antisense ODNs were observed among patients' samples. Incubation of purified myeloma plasma cells with G3139, but not solvent or reverse polarity control ODNs, resulted in a reduction (>75%) of Bcl-2 mRNA levels after 2 and 4 days, as measured by Real-Time PCR. Treatment with G3139 led to a sequence-specific reduction of Bcl-2 protein levels within 4 days of exposure in 10 out of 11 clinical samples from patients with chemosensitive and multidrug-resistant disease, without significant reduction of alpha-Actin, Bax, Bcl-XL, or Mcl-1 proteins. This resulted in a significantly enhanced sensitivity of the myeloma tumor cells to dexamethasone or doxorubicin-induced apoptosis. G3139 can consistently enter myeloma cells, downregulate the expression of Bcl-2, and enhance the efficacy of myeloma therapy. These data support further clinical evaluation of G3139 therapy in multiple myeloma.
一种与Bcl-2 mRNA的前六个密码子互补的反义寡脱氧核苷酸（ODN），G3139（奥布利默森钠；Genasense），已被证明可下调Bcl-2并在包括耐药淋巴瘤在内的多种恶性肿瘤中产生反应。用羧基荧光素（FAM）标记的反义ODN孵育来自多发性骨髓瘤（MM）患者的体外纯化浆细胞，导致其在细胞质和细胞核中的摄取呈时间和剂量依赖性。在患者样本中未观察到Bcl-2反义ODN摄取的主要差异。用G3139而非溶剂或反义对照ODN孵育纯化的骨髓瘤浆细胞，通过实时PCR检测，2天和4天后Bcl-2 mRNA水平降低（>75%）。在来自化疗敏感和多药耐药疾病患者的11个临床样本中的10个中，用G3139处理在暴露4天内导致Bcl-2蛋白水平序列特异性降低，而α-肌动蛋白、Bax、Bcl-XL或Mcl-1蛋白未显著降低。这导致骨髓瘤肿瘤细胞对地塞米松或阿霉素诱导的凋亡敏感性显著增强。G3139可持续进入骨髓瘤细胞，下调Bcl-2表达，并增强骨髓瘤治疗效果。这些数据支持对G3139治疗多发性骨髓瘤进行进一步的临床评估。
