Generation of a lentiviral vector system to efficiently express bioactive recombinant human prolactin hormones

Publication date: Available online 30 September 2019Source: Molecular and Cellular EndocrinologyAuthor(s): Hernán Alarcón, Elena Bonzon-Kulichenko, Rocío Peinado, Filip Lim, Jesús Vázquez, Antonio RodríguezAbstractThe contribution of the pleiotropic hormone Prolactin (PRL) to several physiological and pathological processes is still unknown. To clarify the role of PRL in these processes during the last decade, different human PRL antagonists have been produced to either partially or fully block the wild type hormone activity. In this work, we have cloned these wild type and antagonist sequences in lentivectors (LV) to express them as recombinant self-processing polypeptides by employing a P2A sequence (hPRL-P2A-GFP). We show that these LVs can efficiently transduce and express the hPRL proteins in different cell types and that the P2A sequence does not affect their activities. Additionally, we have tested their activities in paracrine and autocrine cell culture experiments. Our results demonstrate that these recombinant hPRL-P2A proteins are bioactive in both paracrine and autocrine modes, highlighting the potential usefulness of these hPRL-containing LVs for determining the contribution of hPRL to different biological processes.Graphical abstract
Source: Molecular and Cellular Endocrinology - Category: Endocrinology Source Type: research