Effect of YB-1 gene knockdown on human leukemia cell line K562/A02

• Published in: Zhonghua yi xue yi chuan xue za zhi Vol. 26; no. 4; p. 400
• Main Authors: Xu, Wen-lin, Zhou, Lei-lei, Chen, Qiao-yun, Chen, Chen, Fang, Li-li, Fang, Xin-jian, Shen, Hui-ling
• Format: Journal Article
• Language: Chinese
• Published: China 01.08.2009
• Subjects: Apoptosis; Cell Cycle; Cell Line, Tumor; Cell Proliferation; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Leukemia - genetics; Leukemia - metabolism; Leukemia - physiopathology; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; RNA, Small Interfering - genetics; Y-Box-Binding Protein 1
• ISSN: 1003-9406

To investigate the potential effects of YB-1 gene knockdown on gene expression profile, cell growth and apoptosis in leukemia cell line K562/A02. The recombinant eukaryotic expression plasmid containing YB-1 short hairpin RNA (shRNA) or random-sequence (HK) were transfected into K562/A02 cells by lipofectamine mediation. cDNA microarray was performed to explore the alteration of gene expression profile when YB-1 gene expression was decreased. Expression of CARD8 and RHOC genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). The proliferative ability of the cells was determined by methyl thiazolyltetrazolium (MTT) assay and cell cycle analysis. Cell apoptosis was assayed by Annexin V-FITC/PI double labeled flow cytometry. The levels of YB-1 mRNA and protein decreased dramatically in three positively transfected cells when compared with untransfected K562/A02 cells or K562/A02-HK thansfected cells. Gene expression profile was altered by transfection of YB-1 shRNA into K562/A02 cells. Amon