Sosa-García Concepción, Sánchez-Pacheco Uriel Abdallah, Tavira-Montalvan Carlos Alberto, Meneses-Acosta Angélica
Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmacy, Autonomous University of the State of Morelos, Cuernavaca, Morelos, Mexico.
Biomed Res Int. 2025 Feb 25;2025:5556353. doi: 10.1155/bmri/5556353. eCollection 2025.
Recombinant proteins represent one of the greatest achievements of modern pharmaceutical biotechnology, as they are increasingly used across nearly all branches of medicine to treat a wide range of conditions. In response to this demand, various cell engineering approaches have been developed to improve their expression. Some of these approaches involve the use of genetic elements that prevent the silencing of the gene of interest, as well as the generation of resistant cell lines to inhibit or avoid programmed cell death (PCD). This research focuses on analyzing the effects of overexpression of UCOE elements and the HSP27 protein, both individually and together, on the production of human rIFN in HEK293 cells. Our results show that 4-Kb UCOE elements have no effect on protein production in HEK293 cells, while overexpression of HSP27 prolongs the stationary phase during growth kinetics. The Qp of rIFN is 96-fold higher in clones containing the HSP27/UCOE combination compared to the clone containing only UCOE elements or to the control HEK293 cells. These results correlate with the MCP analyses, which showed that overexpression of HSP27 decreased the expression of Bax, caspase 3, cytochrome C, Beclin, and LC3II mRNA. Finally, this study suggests the potential utility of a cell engineering approach based on the overexpression of the human HSP27 protein for enhancing the production of recombinant viruses and proteins in HEK293 cells.
重组蛋白是现代药物生物技术最伟大的成就之一，因为它们越来越多地被应用于几乎所有医学分支，以治疗各种病症。为了满足这一需求，人们开发了各种细胞工程方法来提高它们的表达。其中一些方法涉及使用防止目的基因沉默的遗传元件，以及产生抗性细胞系以抑制或避免程序性细胞死亡（PCD）。本研究着重分析UCOE元件和HSP27蛋白单独及共同过表达对人rIFN在HEK293细胞中产生的影响。我们的结果表明，4-Kb UCOE元件对HEK293细胞中的蛋白产生没有影响，而HSP27的过表达延长了生长动力学中的稳定期。与仅含有UCOE元件的克隆或对照HEK293细胞相比，含有HSP27/UCOE组合的克隆中rIFN的比生产率（Qp）高96倍。这些结果与MCP分析相关，MCP分析表明HSP27的过表达降低了Bax、caspase 3、细胞色素C、Beclin和LC3II mRNA的表达。最后，本研究表明基于人HSP27蛋白过表达的细胞工程方法在增强HEK293细胞中重组病毒和蛋白产生方面的潜在效用。
