Journal info
|
||||
Select Journal
Journals
Bratislava Medical Journal 2024 Ahead of print 2023 2022 2021 2020 2019 2018 2017 2016 2015 2014 2013 2012 Endocrine Regulations General Physiology and Biophysics Neoplasma Acta Virologica Studia Psychologica Cardiology Letters Psychológia a patopsych. dieťaťa Kovove Materialy-Metallic Materials Slovenská hudbaWebshop Cart
Your Cart is currently empty.
Info: Your browser does not accept cookies. To put products into your cart and purchase them you need to enable cookies.
Bratislava Medical Journal Vol.117, No.5, p.272-275, 2016 |
||
Title: Construction of a Hep-2 cell line stably transfected with Livin shRNA | ||
Author: S. L. Wang, W. T. Deng, G. F. Wen, C. W. Li, Y. J. Zeng | ||
Abstract: OBJECTIVE: The aim of this study was to construct a eukaryotic expression plasmid with a short hairpin RNA (shRNA) targeting Livin in order to obtain a stably transfected Hep-2 cell line with a reduced expression of Livin. METHODS: The shRNA targeting Livin mRNA was designed, and a shRNA plasmid and a negative control plasmid were constructed. After amplification in E. coli, restriction endonuclease digestion and sequence confirmation, the plasmids were transfected into Hep-2 cells using Lipofectamine 2000. The stably transfected cell line was screened using G418, and inhibition of Livin mRNA and protein levels were detected using real-time PCR and western blotting, respectively. RESULTS: pGenesil-Livin-shRNA eukaryotic expression plasmid was successfully constructed and identified by sequencing. Green fluorescent protein (GFP) expression was observed in Hep-2 cells transfected with shRNA plasmids by fluorescence microscopy. The expression levels of Livin mRNA and protein decreased significantly in Hep-2 cells transfected with the shRNA recombinant plasmid. The mRNA level was reduced by 47.17 %, and the protein level was reduced by 34.25 %. CONCLUSION: The shRNA eukaryotic expression plasmid targeting Livin was successfully constructed, which could significantly inhibit the expression of Livin in laryngeal cancer Hep-2 cells. This provides a basis for future research on the function of Livin in Hep-2 cells, and gene therapy for laryngeal cancer (Fig. 5, Ref. 10). |
||
Keywords: apoptosis regulatory proteins, cell culture, RNA interference, livin gene | ||
Published online: 13-May-2016 | ||
Year: 2016, Volume: 117, Issue: 5 | Page From: 272, Page To: 275 | |
doi:10.4149/BLL_2016_053 |
||
|
download file |
|