An in vitro human melanoma cell assay was used to work up the partial purification of (a) low molecular weight (MW) substance(s) from aqueous extracts of ovine pineal tissue shown to contain a growth-inhibiting activity. A combination of paper chromatography, ion-exchange and reversephase high performance liquid chromatography with post-column antitumor assay has been developed. This allows a specific identification of an ovine pineal factor (MW<500) which inhibits the growth of human melanoma cells in vitro. The substance was partially purified to about 1,000 times as compared to the IC100-value of the starting material (retentate5). The growth inhibition of human melanoma cells in culture was complete at a dose of 0.1 μg/ml of purified pineal factor(s). It was demonstrated that the activity of this pineal compound differs from some substances known to be present in the pineal, such as melatonin, serotonin, peridines and β-carbolines. The activity was not destroyed by treatment with proteolytic enzymes.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Abbasi A, Ali R, Zaidi ZH (1980) Effective solvent systems for separation and an improved detection method for amino acids by paper chromatography. J Biochem Biophys Methods 3: 311–313
Allen G (1981) In: Work TS, Burdon RH (eds) Laboratory techniques in biochemistry and molecular biology. Elsevier Biomed Press, Oxford, pp 161–236
Aubert C, Prade M, Bohoun C (1970) Effet de la pinéalectomie sur les tumeurs mélaniques de Hamster doré induites par l'administration (per os) d'une seule dose de 9,10-diméthyl-1,2-benzanthracène. C R Acad Sci (Paris) 271: 2465–2466
Bartsch H, Bartsch C (1981) Effect of melatonin on experimental tumors under different photoperiods and times of administration. J Neural Transm 52: 269–279
Bartsch H, Bartsch C, Flehmig B (1986) Differential effect of melatonin on slow and fast growing passages of a human melanoma cell line. Neuroendocrinol Lett 8: 289–293
Bartsch H, Bartsch C, Noteborn HPJM, Flehmig B, Ebels I, Salemink CA (1987) Growth-inhibiting effect of crude pineal extracts on human melanoma cellsin vitro is different from that of known synthetic pineal substances. J Neural Transm 69: 299–311
Bindoni M (1971) Relationship between the pineal gland and the mitotic activity of some tissues. Arch Sci Biol 55: 3–21
Bindoni M, Jutisz M, Ribot G (1976) Characterization and partial purification of a substance in the pineal gland which inhibits cell multiplicatonin vivo. Biochim Biophys Acta 437: 577–588
Blask DE (1984) The pineal: an oncostatic gland? In: Reiter RJ (ed) The pineal gland. Raven Press, New York, pp 253–284
Carr DB, Axelrod L (1981) The pineal gland, pineal dysfunction and cancer. Rev Endocrine-related Canc 8: 13–21
Das Gupta TK, Terz J (1967) Influence of pineal gland on the growth and spread of melanoma in the hamster. Cancer Res 27: 1306–1311
Dilman VM, Anisimov VN, Ostroumova MN, Morosov VG, Khavinson VKh, Azarova MA (1979) Study of anti-tumor effect of polypeptide pineal extract. Oncology 36: 274–280
Ebels I (1980) A survey of the location, isolation and identification of indoles, pteridines and some unknown active substances in sheep pineals. The possible significance of pteridines for the neuroendocrine control of neoplastic growth. J Neural Transm 49: 87–105
El-Domeiri AAH, Das Gupta TK (1976) The influence of pineal ablation and administration of melatonin on growth and spread of hamster melanoma. J Surg Oncol 8: 197–205
Lapin V (1974) Influence of simultaneous pinealectomy and thymectomy on the growth and formation of metastases of the Yoshida sarcoma in rats. Exp Pathol 9: 108–112
Lapin V (1976) Pineal gland and malignancy. Österr Z Onkol 3: 51–59
Lapin V, Ebels I (1976) Effects of some low molecular weight sheep pineal fractions and melatonin on different tumors in rats and mice. Oncology 33: 110–113
Lowry OH, Rosenbrough MJ, Farr AL, Randall (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275
Neifeld JP, Lippman ME (1980) Steroid hormone receptors in melanoma. J Invest Dermatol 74: 379–381
Noteborn HPJM, Ebels I, Pévet P, Reinharz AC, Neascu C, Salemink CA (1982) Comparison of some peptidic and proteic ovine pineal fractions with a bovine pineal E5 fraction. J Neural Transm 55: 27–44
Peterson GL (1977) A simplification of the protein assay method of Lowry et al., which is more generally applicable. Anal Biochem 83: 346–356
Quay WB (1974) Temporal mitotic patterns around a brain lesion; cellular and regional asynchronisms and an effect of pinealectomy. Chronobiologia 1: 237–258
Rampen FHJ, Mulder JH (1980) Malignant melanoma: an androgen-dependent tumour. Lancet 1: 562–565
Rao AP, Rao PN (1976) The cause of G2-arrest in Chinese hamster ovary cells treated with anticancer drugs. J Natl Cancer Inst 57: 1139–1143
Shaw HM, Milton GW, Farago G, McCarthy WH (1978) Endocrine influences on survival from malignant melanoma. Cancer 42: 669–677
Vaughan GM, Reiter RJ, Siler-Khodr TM, Sackman JW, Allen JP, Vaughan MK, McGuire WL, Johnson LY, Starr P (1978) Influence of pinealectomy on serum estrogen and progesterone levels in blind-anosmic female rats. Experientia 34: 1378–1779
Supported in part by grant no. 28-920 of the Praeventiefonds, The Hague, The Netherlands.
About this article
Cite this article
Noteborn, H.P.J.M., Bartsch, H., Bartsch, C. et al. Partial purification of (a) low molecular weight ovine pineal compound(s) with an inhibiting effect on the growth of human melanoma cells in vitro. J. Neural Transmission 73, 135–155 (1988). https://doi.org/10.1007/BF01243385
- Growth inhibition
- melanoma cells
- anti-tumor factor
- ovine pineal