Recombinant adenovirus of microRNA-193b inhibits proliferation of K562 cells

• Published in: Xi bao yu fen zi mian yi xue za zhi Vol. 31; no. 4; p. 452
• Main Authors: Shu, Yan, Qi, Jieyu, Wang, Canwei, Yu, Shujing, Tao, Kun
• Format: Journal Article
• Language: Chinese
• Published: China 01.04.2015
• Subjects: Adenoviridae - genetics; Adenoviridae - metabolism; Cell Proliferation; Down-Regulation; HEK293 Cells; Humans; K562 Cells; Leukemia, Erythroblastic, Acute - genetics; Leukemia, Erythroblastic, Acute - metabolism; Leukemia, Erythroblastic, Acute - physiopathology; MicroRNAs - genetics; MicroRNAs - metabolism
• ISSN: 1007-8738

To construct a recombinant adenovirus vector containing human pre-miR-193b and investigate its effect on the proliferation of chronic myelocytic leukemia K562 cells. The cDNA of pre-miR-193b was obtained by chemical synthesis and inserted into the adenoviral shuttle vector pAdTrack-CMV. The recombinant shuttle plasmid was linearized by Pme I and transformed into AdEasier cells for homologous recombination in E.coli BJ5183 with the adenoviral backbone plasmid pAdEasy-1. The recombinant adenoviral plasmid was linearized by Pac I and then used for transfecting HEK293 cells. After package and amplification in HEK293 cells, the virus titer was determined by serial dilution assay. The expression level of miR-193b in K562 cells was detected by real-time quantitative PCR. The cell proliferation was observed by MTT assay. The recombinant plasmid named pAd-miR-193b was confirmed by restriction enzyme analysis and sequencing. The recombinant adenovirus could infect K562 cells efficiently. Compared with control group, re