The Study of Recombinant Interfering Lentiviruses and Overex-pressed Adenovirus Vectors Targeting Human c-Cbl Gene:Con-struction,Identification and Efficacy
10.19746/j.cnki.issn1009-2137.2024.01.044
- VernacularTitle:靶向人c-Cbl基因重组干扰慢病毒与过表达腺病毒载体的构建、鉴定以及病毒功效研究
- Author:
Qi-Xin SUN
1
;
Bing-Yi WU
;
Qian-Qian YAO
;
Zhi-Wei HUANG
;
Zhi-Gang ZHU
Author Information
1. 广州市第一人民医院(华南理工大学第二附属医院)老年血液肿瘤科,广东广州 510180
- Keywords:
c-Cbl gene;
lentivirus vectot;
adenovirus vector
- From:
Journal of Experimental Hematology
2024;32(1):274-281
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct recombinant lentivirus and adenovirus which regulate the expression of c-Cbl gene and evaluate their efficacy.Methods:The interference lentivirus and overexpressed adenovirus targeting human c-Cbl gene were constructed by gene recombination technology.Quantitative PCR and western blotting were used to detect the expression changes in c-Cbl gene and its transcription after leukemia cells(HL60,THP1)were infected by virus.Results:Three recombinant interfering lentiviral vectors targeting human c-Cbl genes to successfully constructed and were identified by DNA sequencing,and the titers of the packaged viruses were all greater than 1 x 108 TU/ml.Among them,shRNA-2 lentivirus had the highest interference efficiency,and the expression of c-Cbl gene and CBL protein were decreased about 95％and 60％respectively after leukemia cells were infected with shRNA-2;In addition,the recombinant overexpression adenovirus targeting human c-Cbl gene was packaged successfully with the virus titer greater than 1 x 109 TU/ml.When leukemia cells were infected with adenovirus,the expression of c-Cbl gene and CBL protein were up-regulated about 10 times and 1.5 times respectively.Conclusion:Both recombinant interfering lentivirus and overexpression adenovirus can efficiently infect leukemia cells and affect the expressions of c-Cbl gene and CBL protein.It will lay a preliminary foundation for the subsequent study on the function of c-Cbl gene in tumor cells.
