Baglia L A, Cruz D, Shaw J E
Department of Medical Microbiology and Immunology, College of Medicine Ohio State University, Columbus 43210-1239.
Endocrinology. 1991 May;128(5):2266-72. doi: 10.1210/endo-128-5-2266.
Pituitary human PRL (hPRL) antiserum inhibits growth of B-lymphoblastoid cells in vitro, but the mechanism of inhibition is unclear. In this study the mechanism of inhibition of human B-cell growth by anti-hPRL was explored with an Epstein-Barr virus nuclear antigen (EBNA)-negative Burkitt lymphoma cell line (sfRamos) that proliferates continuously in serum-free medium with human transferrin as the only protein supplement. The data show that antiserum immunoglobulin fraction G (IgG) to pituitary hPRL, but not nonimmune serum IgG, completely inhibited the growth of sfRamos cells. In addition, anti-hPRL IgG identified a single band (29 kDa) in sfRamos spent medium, but not in fresh serum-free medium or in human transferrin, as demonstrated by sodium dodecyl sulfate-reducing polyacrylamide gel electrophoresis and Western immunoblot analysis. Polyacrylamide gel electrophoresis and Western analysis of a mixture containing sfRamos spent medium and excess pituitary hPRL established that the sfRamos 29-kDa PRL-like protein (PLP29) and pituitary hPRL (23 kDa) were electrophoretically distinct. Finally, sfRamos spent medium, but not fresh serum-free medium, was mitogenic for sfRamos and Nb2, a PRL-sensitive node rat lymphoma cell line. These findings demonstrate that PLP29 is biologically and immunologically like pituitary hPRL, but is electrophoretically different from this hormone. We suggest that PLP29 is secreted as an autocrine growth factor by sfRamos Burkitt lymphoma cells during continuous serum-free growth.
垂体人催乳素(hPRL)抗血清在体外可抑制B淋巴母细胞的生长,但抑制机制尚不清楚。在本研究中,我们使用一种爱泼斯坦-巴尔病毒核抗原(EBNA)阴性的伯基特淋巴瘤细胞系(sfRamos),该细胞系在以人转铁蛋白作为唯一蛋白质补充剂的无血清培养基中持续增殖,以此来探索抗hPRL抑制人B细胞生长的机制。数据显示,垂体hPRL的抗血清免疫球蛋白G(IgG)组分可完全抑制sfRamos细胞的生长,而非免疫血清IgG则无此作用。此外,通过十二烷基硫酸钠还原聚丙烯酰胺凝胶电泳和Western免疫印迹分析表明,抗hPRL IgG在sfRamos的消耗培养基中鉴定出一条单一的条带(29 kDa),但在新鲜无血清培养基或人转铁蛋白中未检测到。对含有sfRamos消耗培养基和过量垂体hPRL的混合物进行聚丙烯酰胺凝胶电泳和Western分析,结果表明sfRamos的29 kDa催乳素样蛋白(PLP29)和垂体hPRL(23 kDa)在电泳上是不同的。最后,sfRamos消耗培养基对sfRamos和Nb2(一种PRL敏感的大鼠淋巴瘤细胞系)具有促有丝分裂作用,而新鲜无血清培养基则无此作用。这些发现表明,PLP29在生物学和免疫学上与垂体hPRL相似,但在电泳上与该激素不同。我们认为,PLP29是sfRamos伯基特淋巴瘤细胞在持续无血清生长过程中作为自分泌生长因子分泌的。