Purification and Characterization of New Mammary Cell Growth Factor from Rat Pituitary Tumors : Preliminary Report.

大鼠垂体肿瘤中新型乳腺细胞生长因子的纯化和表征：初步报告。

• 批准号: 01570193
• 负责人: KAMIYA Kenji
• 金额: $ 1.34万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570193/
• 关键词: Rat mammary epithelial cell line; Rat pituitary cell line; Autocrine growth factor; Glucocorticoid; Chemically defined medium; 成長因子; 乳腺上皮細胞; 下垂体由来増殖因子; 乳腺由来増殖因子

Normal rat mammary epithelial cell line was established from continuous cultures of rat mammary cells in chemically defined medium. These cells were shown to be mammary epithelial origin by in vivo transplantation assay, electron microscopic examination and immunocytochemical analysis. Using these mammary cells as target cells, in vitro bioassay system for the detection of the mitogenic activity on mammary cells was developed.Four rat pituitary cell lines, derived from mammotropic pituitary tumors (MtT-F4, MtT-F84), and radiation induced somatomammotropic pituitary tumors (PT1,PT3), were also established in chemically defined medium. As the results of in vitro analysis of mitogenic activity of conditioned medium from established pituitary cell cultures, conditioned medium from PT3 was shown to have strong growth-promoting effects on mammary cells. cc152 cells, isolated from PT3, were found to have unique growth characteristics that cc152 cells could proliferate in the chemically defined medium supplemented with insslin and hydrocortisone without any other growth-promoting factors. Furthermore, cc152-c3 cells derived from cc152 were able to proliferate in the chemically defined medium supplemented with hydrocortisone alone. Conditioned medium from cc152-c3 cells stimulated the growth of their own cells. The growth-stimulating effects of hydrocortisone on cc152 and cc152-c3 cells was inhibited by the addition of a glucocorticoid receptor antagonist, RV486. RV486 also blocked hydrocortisone-induced growth-stimulating effects of conditioned medium. These results indicated that stimulation of cc152-c3 cells with hydrocortisone resulted in secretion of an autocrine growth factor (s).A preliminary characterization of the growth factor (s) in the conditioned medium suggested that growth factor (s) was a polypeptide. Experiments are being carried out to purify and further characterize the glucocorticoid-induced growth factor (s).

正常大鼠乳腺上皮细胞系是通过在化学成分确定的培养基中连续培养大鼠乳腺细胞而建立的。通过体内移植试验、电子显微镜检查和免疫细胞化学分析表明这些细胞是乳腺上皮来源的。以这些乳腺细胞为靶细胞，开发了检测乳腺细胞促有丝分裂活性的体外生物测定系统。四种大鼠垂体细胞系，源自促乳腺垂体瘤（MtT-F4、MtT-F84）和辐射诱导的促体乳腺瘤垂体瘤（PT1、PT3）也在化学成分确定的培养基中建立。对已建立的垂体细胞培养物的条件培养基的促有丝分裂活性进行体外分析的结果表明，PT3 的条件培养基对乳腺细胞具有强烈的生长促进作用。从PT3中分离的cc152细胞被发现具有独特的生长特性，cc152细胞可以在补充有insslin和氢化可的松且不含任何其他生长促进因子的化学成分确定的培养基中增殖。此外，源自 cc152 的 cc152-c3 细胞能够在仅补充有氢化可的松的化学成分确定的培养基中增殖。 cc152-c3 细胞的条件培养基刺激了其自身细胞的生长。添加糖皮质激素受体拮抗剂 RV486 可以抑制氢化可的松对 cc152 和 cc152-c3 细胞的生长刺激作用。 RV486 还阻断氢化可的松诱导的条件培养基的生长刺激作用。这些结果表明，用氢化可的松刺激cc152-c3细胞导致自分泌生长因子的分泌。条件培养基中生长因子的初步表征表明生长因子是多肽。正在进行实验以纯化和进一步表征糖皮质激素诱导的生长因子。

项目成果

Kamiya,K.,Gould,M.N.&Clifton,K.H.: "Differential control of alveolar and ductal development in grafts of monodispersed rat mammary epithelium." Proc.Soc.Exp.Biol.Med.196. 284-292 (1991)

神谷，K.，古尔德，M.N.

Kamiya,K.,Kim,N.D.,Gould,M.N.&Clifton,K.H.: "Repair of potentially lethal damages in rat mammary clonogens following irradiation in organoid culture." Int.J.Radiat.Biol.

神谷，K.，金，N.D.，古尔德，M.N.

Kamiya,K.,Gould,M.N.& Clifton,K.H.: "Differential control of alveolar and development in grafts of monodispersed rat mammary epithelium." Proc.Soc.Exp.Biol.Med.196. 284-292 (1991)

神谷，K.，古尔德，M.N.

Kamiya,K.,Yokoro,K.,et al.: "Clonogenic cells and rat mammary cancer:Effects of hormones,X rays,and fission neutrons" Radiation Research. 120. 323-338 (1989)

Kamiya,K.、Yokoro,K. 等人：“克隆细胞和大鼠乳腺癌：激素、X 射线和裂变中子的影响”辐射研究。

Kamiya, K., Gould, M. N. and Clifron, K. H.: "Differential control of alveolar and ductal development in grafts of monodispersed rat mammary epithelium." Proc. Soc. Exp. Biol. Med.196. 284-292 (1991)

Kamiya, K.、Gould, M. N. 和 Clifron, K. H.：“单分散大鼠乳腺上皮移植物中肺泡和导管发育的差异控制。”