Background:NOTCH1 mutation is an essential molecular biologic aberration in chronic lymphocytic leukemia(CLL).CLL patients withNOTCH1 mutation have shown an unfavorable survival and a poor response to chemoimmunothera...Background:NOTCH1 mutation is an essential molecular biologic aberration in chronic lymphocytic leukemia(CLL).CLL patients withNOTCH1 mutation have shown an unfavorable survival and a poor response to chemoimmunotherapy.This study aims to present the mechanisms of adverse prognosis caused byNOTCH1 mutation from the perspective of the splicing factor heterogeneous nuclear ribonucleoprotein A1(hnRNPA1).Methods:The microarray data in Gene Expression Omnibus datasets were analyzed by bioinformatics and the function of hnRNPA1 was checked by testing the proliferation and apoptosis of CLL-like cell lines.Afterward,quantitative reverse transcription-polymerase chain reaction and Western blotting were applied to explore the relationship among NOTCH1,c-Myc,and hnRNPA1.Results:RNA splicing was found to play a vital part inNOTCH1-mutated CLL cells;hence,hnRNPA1 was selected as the focus of this study.Higher expression of hnRNPA1 validated in primaryNOTCH1-mutated CLL samples could promote proliferation and inhibit apoptosis in CLL.The expression of hnRNPA1 increased when NOTCH1 signaling was activated by transfection with NOTCH1 intracellular domain(NICD)-overexpressed adenovirus vector and declined after NOTCH1 signaling was inhibited by NOTCH1-shRNA.Higher expression of c-Myc was observed in NICD-overexpressed cells and hnRNPA1 expression was downregulated after applying c-Myc inhibitor 10058-F4.Moreover,in NICD-overexpressed cells,hnRNPA1 expression decreased through c-Myc inhibition.Conclusion:Overexpression of c-Myc-dependent hnRNPA1 could promote proliferation and inhibit apoptosis inNOTCH1-mutated CLL cells,which might partly account for the poor prognosis of patients withNOTCH1 mutation.展开更多
基金National Natural Science Foundation of China(Nos.81700193,81970146,82170186,and 81720108002)National Major Science and Technology Projects of China(No.2018ZX09734-007)+1 种基金China Postdoctoral Science Foundation(No.2021M691336)Jiangsu Postdoctoral Science Foundation(No.2021K083A)。
文摘Background:NOTCH1 mutation is an essential molecular biologic aberration in chronic lymphocytic leukemia(CLL).CLL patients withNOTCH1 mutation have shown an unfavorable survival and a poor response to chemoimmunotherapy.This study aims to present the mechanisms of adverse prognosis caused byNOTCH1 mutation from the perspective of the splicing factor heterogeneous nuclear ribonucleoprotein A1(hnRNPA1).Methods:The microarray data in Gene Expression Omnibus datasets were analyzed by bioinformatics and the function of hnRNPA1 was checked by testing the proliferation and apoptosis of CLL-like cell lines.Afterward,quantitative reverse transcription-polymerase chain reaction and Western blotting were applied to explore the relationship among NOTCH1,c-Myc,and hnRNPA1.Results:RNA splicing was found to play a vital part inNOTCH1-mutated CLL cells;hence,hnRNPA1 was selected as the focus of this study.Higher expression of hnRNPA1 validated in primaryNOTCH1-mutated CLL samples could promote proliferation and inhibit apoptosis in CLL.The expression of hnRNPA1 increased when NOTCH1 signaling was activated by transfection with NOTCH1 intracellular domain(NICD)-overexpressed adenovirus vector and declined after NOTCH1 signaling was inhibited by NOTCH1-shRNA.Higher expression of c-Myc was observed in NICD-overexpressed cells and hnRNPA1 expression was downregulated after applying c-Myc inhibitor 10058-F4.Moreover,in NICD-overexpressed cells,hnRNPA1 expression decreased through c-Myc inhibition.Conclusion:Overexpression of c-Myc-dependent hnRNPA1 could promote proliferation and inhibit apoptosis inNOTCH1-mutated CLL cells,which might partly account for the poor prognosis of patients withNOTCH1 mutation.
