Bindoni M, Jutisz M, Ribot G
Biochim Biophys Acta. 1976 Jul 21;437(2):577-88. doi: 10.1016/0304-4165(76)90025-8.
A substance which inhibits the in vitro multiplication of 3 cell strains, 37 RC, KB and NCTC clone 929, was characterized in the sheep pineal gland and partially purified using three successive chromatography techniques, respectively on Sephadex G-25, CM-cellulose and Biogel P 60 columns. The sheep cerebral cortex and liver also contain, but at much lower concentration than in the pineal, substance(s) that behave in different tests like the factor isolated from the pineal. The nature of the antimitotic substance from the pineal is as yet unknown. It is not destroyed by treatment with proteolytic enzymes, nor by boiling with 6 M HC1. It was established that it is different from the known antiblastic drugs such as Daunomycin and Methotrexate and from some active substances known to be present in the pineal, such as melatonin, secotonin and norepinephrine, which, in the same conditions, did not show any antimitotic activity. It was shown that when the concentration of the pineal factor in the culture medium was high enough (10 mug/ml), the inhibition of the KB cells multiplication was complete and irreversible. Microscopic examination of the treated cells showed that the morphological alteration was rapid (3--6 h) and deep, with shrinkage of both cytoplasm and nucleus, while with antiblastic drugs, morphological alteration proceeded slower (1--3 days) and appeared less pronounced.
一种能抑制3种细胞株（37 RC、KB和NCTC克隆929）体外增殖的物质在绵羊松果体中得到鉴定，并分别采用Sephadex G - 25、CM - 纤维素和Biogel P 60柱上的三种连续色谱技术进行了部分纯化。绵羊大脑皮层和肝脏中也含有与从松果体中分离出的因子在不同试验中表现相似的物质，但浓度比松果体中的低得多。来自松果体的抗有丝分裂物质的性质尚不清楚。它不会被蛋白水解酶处理或用6 M HCl煮沸破坏。已确定它不同于已知的抗瘤药物如柔红霉素和甲氨蝶呤，也不同于已知存在于松果体中的一些活性物质，如褪黑素、5 - 羟色胺和去甲肾上腺素，这些物质在相同条件下未显示出任何抗有丝分裂活性。结果表明，当培养基中松果体因子的浓度足够高（10微克/毫升）时，对KB细胞增殖的抑制是完全且不可逆的。对处理过的细胞进行显微镜检查发现，形态改变迅速（3 - 6小时）且明显，细胞质和细胞核均收缩，而使用抗瘤药物时，形态改变进展较慢（1 - 3天）且不太明显。
