Furui T, Imai A, Ochi N, Tamaya T
Department of Obstetrics and Gynecology, Gifu University School of Medicine, Japan.
Cancer. 1994 Feb 15;73(4):1239-44. doi: 10.1002/1097-0142(19940215)73:4<1239::aid-cncr2820730418>3.0.co;2-y.
Uterine endometrial carcinoma has been reported to synthesize and secrete some putative mitogens that elicit either a positive or negative proliferation response in endometrial fibroblasts. The purposes of this study were to isolate and to identify the negative growth factor(s) from endometrial carcinoma extract.
The factor was isolated by a sequence of molecular size exclusion filtration, anion exchange chromatography, gel filtration and Affi-Gel Blue chromatography, followed by NH2-terminal amino acid sequencing. Mitogenicity was determined by [3H]thymidine incorporation into the endometrial fibroblasts.
The purification procedure yielded a single active protein band (68 kDa). The protein, purified approximately 20,000-fold, evoked 90% inhibition of [3H]-thymidine incorporation into endometrial fibroblasts in the nanomolar range. This potent growth inhibitor is a previously unidentified protein molecule as revealed by amino acid sequences.
Endometrial carcinoma could produce a new protein that may act as a paracrine factor to suppress the growth of its stroma endometrial fibroblasts.
据报道，子宫内膜癌可合成并分泌一些假定的有丝分裂原，这些有丝分裂原可引起子宫内膜成纤维细胞的增殖反应，反应结果可能为阳性或阴性。本研究的目的是从子宫内膜癌提取物中分离并鉴定负性生长因子。
通过一系列分子大小排阻过滤、阴离子交换色谱、凝胶过滤和Affi-Gel蓝色谱法分离该因子，随后进行氨基末端氨基酸测序。通过将[3H]胸苷掺入子宫内膜成纤维细胞来测定有丝分裂活性。
纯化过程产生了一条单一的活性蛋白带（68 kDa）。该蛋白纯化了约20000倍，在纳摩尔范围内可引起[3H]胸苷掺入子宫内膜成纤维细胞的抑制率达90%。氨基酸序列显示，这种强效生长抑制剂是一种此前未被鉴定的蛋白质分子。
子宫内膜癌可产生一种新的蛋白质，该蛋白质可能作为旁分泌因子抑制其基质子宫内膜成纤维细胞的生长。
