Bovine prolactin elevates hTF expression directed by a tissue-specific goat β-casein promoter through prolactin receptor-mediated STAT5a activation

• Published in: Biotechnology letters 2012-11, Vol.34 (11), p.1991-1999
• Main Authors: Jiang, Shizhong, Ren, Zhaorui, Xie, Fei, Yan, Jingbin, Huang, Shuzhen, Zeng, Yitao
• Format: Article
• Language: English
• Subjects: Activation; Analysis of Variance; Animals; antagonists; Applied Microbiology; beta-casein; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; biopharmaceuticals; Biotechnology; Blotting, Western; Caseins - genetics; Cattle; cell culture; Cell Line; Dose-Response Relationship, Drug; epithelial cells; Fundamental and applied biological sciences. Psychology; genes; Goats; Human; Humans; Life Sciences; Mammary glands; messenger RNA; Mice; Microbiology; Milk; Original Research Paper; plasmids; prolactin; Prolactin - genetics; Prolactin - metabolism; Promoter Regions, Genetic; Real-Time Polymerase Chain Reaction; Receptors, Prolactin - antagonists & inhibitors; Receptors, Prolactin - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; STAT5 Transcription Factor - genetics; STAT5 Transcription Factor - metabolism; Transducers; Transfection; Transferrin; Transferrin - biosynthesis; Transferrin - genetics; Transferrin - metabolism; Transgenic; transgenic animals
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-012-1009-1

Prolactin promotes the expression of exogenous human transferrin gene in the milk of transgenic mice. To elucidate this, a recombinant plasmid of bovine prolactin plus human transferrin vector was co-transfected into cultured murine mammary gland epithelial cells. Prolactin-receptor antagonist and shRNA corresponding to prolactin-receptor mRNA were added into the cell culture mixture to investigate the relations between prolactin-receptor and human transferrin expression after bovine prolactin inducement. Levels of human transferrin in the supernatants were increased under the presentation of bovine prolactin (from 1,076 ± 115 to 1,886 ± 114 pg/ml). With the treatment of prolactin-receptor antagonist or shRNA, human transferrin in cells was declined (1,886 ± 113 vs. 1,233 ± 85 pg/ml or 1,114 ± 75 pg/ml, respectively). An inverse correlation was found between the dosage of prolactin-receptor antagonist and expression level of human transferrin. Real-time qRT-PCR analysis showed that the relative level of signal transducer and activator of transcription 5a (STAT5a) transcript in transfected cells correlated with expression levels of human transferrin in the supernatant of the same cells. Bovine prolactin thus improved the expression of human transferrin through such a possible mechanism that bovine prolactin activated STAT5a transcription expression via combined with prolactin-receptor and suggest a potential utility of the bovine prolactin for efficient expression of valuable pharmaceutical proteins in mammary glands of transgenic animals.