Effects Of Gene Silencing Of Aminolevulinic Acid Synthetase MRNA On The Expression Of Protein Band 3 In Erythrocyte

Objectives Our present project is to explore the effects ofeALAS-mRNA is silenced by shRNA on erythroid differentiation of K562 cells.Methods K562 cells were divided into three groups: iron deficiency group,iron-rich group and shRNA (iron-rich) group respectively. Then the cells werecultured 120h with the optimized erythroid differentiation condition. Theexpression of band 3 mRNA and protein were examined by Realtime RT-PCRand Western blot. Results The expression of band 3 mRNA in shRNAtransfection group was lower than iron-rich group. Of note, the lowestexpression was achieved in iron deficiency group. The expression of band3mRNA was consistent with band 3 protein which was detected with Westernblot. Conclusions The shRNA vector targeting eALAS mRNA can efficiently silence the eALAS expression, and reduce eALAS expression inerythroid differentiation of K562 cells could suppress the band 3 proteinexpression. Iron deficiency can markedly reduce the expression band 3 protein.Our study demonstrate for the first time that iron regulates the expression ofband 3 protein through effecting the level of eALAS-mRNA.