Preparation and characterization of Ad-ERα-36-Fc-GFP.
- Author:
Yuqiong XIE
1
;
Chunchun LI
1
;
Xiaoye LI
1
;
Lihong CHEN
1
;
Maoxiao YAN
2
;
Jiang CAO
1
Author Information
- Publication Type:Journal Article
- Keywords: Ad-ERα-36-Fc-GFP; adenovirus packaging; decoy receptor; gene therapy
- MeSH: Adenoviridae/genetics*; Cell Proliferation; Estrogen Receptor alpha/metabolism*; Recombinant Proteins; Transfection
- From: Chinese Journal of Biotechnology 2022;38(3):1086-1095
- CountryChina
- Language:Chinese
- Abstract: ERα-36 is a novel subtype of estrogen receptor α which promotes tumor cell proliferation, invasion and drug resistance, and it serves as a therapeutic target. However, only small-molecule compounds targeting ERα-36 are under development as anticancer drugs at present. Gene therapy approach targeting ERα-36 can be explored using recombinant adenovirus armed with decoy receptor. The recombinant shuttle plasmid pDC316-Ig κ-ERα-36-Fc-GFP was constructed via genetic engineering to express an Ig κ-signaling peptide-leading secretory recombinant fusion protein ERα-36-Fc. The recombinant adenovirus Ad-ERα-36-Fc-GFP was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system. The expression of fusion protein and functional outcome of Ad-ERα-36-Fc-GFP transduction were further analyzed with triple-negative breast cancer MDA-MB-231 cells. Results showed that the recombinant adenovirus Ad-ERα-36-Fc-GFP was successfully generated. The virus effectively infected MDA-MB-231 cells which resulted in expression and secretion of the recombinant fusion protein ERα-36-Fc, leading to significant inhibition of EGFR/ERK signaling pathway. Preparation of the recombinant adenovirus Ad-ERα-36-Fc-GFP provides a basis for further investigation on cancer gene therapy targeting ERα-36.
