Knockdown of ALAS2 Affects Erythroid Differentiation by Down-regulating Mitophagy Receptor BNIP3L

• Published in: Zhongguo shi yan xue ye xue za zhi Vol. 28; no. 5; p. 1710
• Main Authors: Hu, Shu-Ting, Li, Xin-Yu, Chen, Han, Xu, Lyu-Hong, Fang, Jian-Pei
• Format: Journal Article
• Language: Chinese
• Published: China 01.10.2020
• Subjects: 5-Aminolevulinate Synthetase - metabolism; Apoptosis; Carrier Proteins; Cell Differentiation; Humans; K562 Cells; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mitophagy; Proto-Oncogene Proteins - genetics; Tumor Suppressor Proteins
• ISSN: 1009-2137
• DOI: 10.19746/j.cnki.issn.1009-2137.2020.05.047

AbstractObjective: To investigate the effect of ALAS2 downregulation on the expression of BNIP3L and erythroid differentiation in K562 cells. The expression of ALAS2 was down-regulated by transfection of lentivirus, then quantitative real-time PCR was performed to detect the transfection efficiency. Flow cytometry analysis was applied to evaluate apoptosis of cells, erythroid differentiation, mitochondrial membrane potential and reactive oxygen species (ROS) level. Western blot was used to detect the BNIP3L expression, Co-immunoprecipitation was performed to analyze the relationship between ALAS2 and BNIP3L. Compared with sh-NC group, knockdown of ALAS2 induced downregulation of BNIP3L mRNA and protein expression(P<0.01) and erythroid related transcription factors GATA1, Nrf2 expression, as well as reduction of ROS level(P<0.05). Mitochondrial membrane potential of control (sh-NC) group was lower than that of shALAS2 group(P<0.05), but there was no significant change of cell apoptotic rate in two groups. CD71