Establishment of Human Embryonic Kidney-239 Cell Line with High-Stress Resistance and Protein Expression Capacity

Background: High yield production of recombinant proteins with proper glycosylation, folding, solubility and bioactivity is one of the most important challenges in the field of bio-phar-maceutical industries. Human embryonic kidney-239 (HEK-239) cell line is used as mammalian expression system for recombinant protein production. Any manipulation to improve ability of this cell line for expression of desired recombinant proteins is great of interest. NFE2L2 gene, as master gene in cellular redox hemostasis, and its transcription factor, nuclear factor E2-related factor-2 (Nrf-2), play important role in cell proliferation and oxidant/ antioxidant balance. Here in we es-tablished Nrf2-overexpressing-stable HEK293 cell line with UV-resistance capacity for future using in term of recombinant protein production.Materials and Methods: HEK-293 cell line was transfected with Nrf2 containing pcDNA3. geneticin was used to select stable clones. RT-PCR and western blot were done to confirm Nrf2 expression after transfection. Water-soluble tetrazolium salts-1 (WST-) assay was performed to determine proliferation rate of UV irradiation stress induced HEK-239. The expression of Quinone Oxidoreductase 1 (NQO1) and certain glutathione S-transferases (GSTs), as Nrf2 downstream genes, was evalu-ated by real-time PCR in UV exposed HEK-293 and HEK-293. Results: Transfected HEK-293 expressed Nrf2 gene at tran-scriptional and translational levels. WST-1 results indicated that the Nrf2-overexpressing HEK293 showed higher prolifera-tion rate even after exposure to UV irradiation in comparison with non-transfected ones. High expression levels of NQO1 and GSTs were detected in transfected and UV stress induced HEK-293.Conclusion: This Nrf2-overexpressing-stable HEK293 with higher growth rate augmented for more resistance against stress condition could use for high yield production of recombinant proteins.

کلیدواژه ها:

HEK-293 Cell Line ، Nrf2 ، Recombinant Protein