Park Juyoung, Franco Robert S, Augsburger James J, Banerjee Rupak K
Department of Mechanical Engineering, University of Cincinnati, Cincinnati, OH 45221-0072, USA.
Curr Eye Res. 2007 Jul-Aug;32(7-8):659-67. doi: 10.1080/02713680701473244.
Methotrexate (MTX) is the most commonly used chemotherapeutic agent to treat primary central nervous system lymphoma (PCNSL) and intraocular lymphoma (IOL). 2-methoxyestradiol (2ME2) is a potent antitumor and anti-angiogenesis agent which, unlike other cytotoxic drugs, has minimal toxicity. In this study, anti-proliferative, apoptotic, and cell-cycle effects of 2ME2 and MTX were compared to evaluate 2ME2 efficacy in human lymphoma cells, models for non-Hodgkin B cell lymphomas.
The cells were cultured and incubated with varying concentrations of 2ME2 or MTX. A tetrazolium-based colorimetric assay was used to quantify the anti-proliferative effects of 2ME2 and MTX using a microplate reader. To detect apoptotic and cell cycle distribution changes induced by 2ME2 and MTX, the cells were stained with Annexin V-FITC and/or propidium iodide (PI) and analyzed by flow cytometry.
Lymphoma cell proliferation was inhibited by 50% at concentrations ranging from 0.4 to 1 microM for 2ME2 and 0.06 to 0.2 microM for MTX. Induction of apoptosis by 2ME2 and MTX was observed in the tested cells. 2ME2 was a G2/M-phase specific blocker whereas MTX was an S-phase specific blocker in cell cycle analyses. At 1 microM concentration, 2ME2 and MTX showed similar anti-proliferative effect on the lymphoma cell lines. In previously reported studies, for normal endothelial cells, 1 microM 2ME2 showed no appreciable toxicity, while MTX at this same concentration exhibited significant cytotoxicity. 2ME2 at a therapeutic target concentration of 1 mu M may be an effective and relatively non-toxic drug for the treatment of PCNSL with IOL.
Our study of the effect of 2ME2 and MTX on anti-proliferation, apoptosis, and cell cycling suggests that 2ME2 is a potential agent for treating PCNSL and IOL.
甲氨蝶呤（MTX）是治疗原发性中枢神经系统淋巴瘤（PCNSL）和眼内淋巴瘤（IOL）最常用的化疗药物。2-甲氧基雌二醇（2ME2）是一种有效的抗肿瘤和抗血管生成药物，与其他细胞毒性药物不同，其毒性极小。在本研究中，比较了2ME2和MTX的抗增殖、凋亡及细胞周期作用，以评估2ME2在人淋巴瘤细胞（非霍奇金B细胞淋巴瘤模型）中的疗效。
培养细胞并用不同浓度的2ME2或MTX进行孵育。使用基于四氮唑的比色法，通过酶标仪定量2ME2和MTX的抗增殖作用。为检测2ME2和MTX诱导的凋亡及细胞周期分布变化，用膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）和/或碘化丙啶（PI）对细胞进行染色，并通过流式细胞术分析。
2ME2浓度在0.4至1微摩尔/升、MTX浓度在0.06至0.2微摩尔/升时，淋巴瘤细胞增殖被抑制50%。在受试细胞中观察到2ME2和MTX诱导凋亡。在细胞周期分析中，2ME2是G2/M期特异性阻滞剂，而MTX是S期特异性阻滞剂。在1微摩尔/升浓度下，2ME2和MTX对淋巴瘤细胞系显示出相似的抗增殖作用。在先前报道的研究中，对于正常内皮细胞，1微摩尔/升的2ME2未显示出明显毒性，而相同浓度的MTX则表现出显著的细胞毒性。治疗靶点浓度为1微摩尔的2ME2可能是治疗伴有IOL的PCNSL的一种有效且相对无毒的药物。
我们对2ME2和MTX抗增殖、凋亡及细胞周期作用的研究表明，2ME2是治疗PCNSL和IOL的潜在药物。
