Lan Zhang1, Weihua Zhang1. 1. Department of Haematology, First Hospital of Shanxi Medical University, Taiyuan 030001, China, suoai707c@163.com.
Abstract
BACKGROUND: NUDT21 is a mammalian precursor mRNA(pre-mRNA) 3' end processing factor and plays an important role in the selection of poly(A) sites in 3'-untranslated region (3'-UTR). NUDT21 links alternative polyadenylation with regulation of glioblastoma and osteosarcoma progression and is found to be related to drug resistance in childhood acute leukemia. However, the effect of NUDT21 on leukemia cells and the underlying mechanism are unknown. METHODS: We knocked down NUDT21 in K562 cells and applied qRT-PCR and western blotting to quantitate the mRNA and protein expression. Cell proliferating and apoptosis were investigated subsequently by flow cytometry, BrdU, Caspase3/7. RNA microarray and intracellular signaling array were used to determine the important cell signaling pathways. RESULTS: We clarified that the mRNA expression levels of NUDT21 are higher in primary chronic myelocytic leukemia patients and K562 leukemic cells compared with healthy controls and PBMCs. Downregulation of NUDT21 expression in K562 cells inhibits proliferation and promotes apoptosis. Screening by mRNA chip and intracellular signaling array, we found that MAPK/ERK pathway represented the main molecular mechanism underlying the effects of NUDT21 knockdown in K562 cells. CONCLUSION: NUDT21 played an important role in promoting proliferation and inhibiting apoptosis in leukemia K562 cells. The underlying mechanisms involved the modulation of PTEN and a set of downstream molecules including ERK1/2. IMPACT STATEMENT: The present work shows that the expression of NUDT21 was upregulated in chronic myelocytic leukemia and K562 cells. Silencing NUDT21 inhibited the proliferation and promoted the apoptosis of K562 cells. Subsequent experiments confirmed that NUDT21 promoted K562 proliferation through regulating the expression of p-ERK. Our findings may provide insights into the molecular mechanism underlying the effects of NUDT21 on leukemia cells and a novel strategy for the treatment of leukemia.
BACKGROUND: NUDT21 is a mammalian precursor mRNA(pre-mRNA) 3' end processing factor and plays an important role in the selection of poly(A) sites in 3'-untranslated region (3'-UTR). NUDT21 links alternative polyadenylation with regulation of glioblastoma and osteosarcoma progression and is found to be related to drug resistance in childhood acute leukemia. However, the effect of NUDT21 on leukemia cells and the underlying mechanism are unknown. METHODS: We knocked down NUDT21 in K562 cells and applied qRT-PCR and western blotting to quantitate the mRNA and protein expression. Cell proliferating and apoptosis were investigated subsequently by flow cytometry, BrdU, Caspase3/7. RNA microarray and intracellular signaling array were used to determine the important cell signaling pathways. RESULTS: We clarified that the mRNA expression levels of NUDT21 are higher in primary chronic myelocytic leukemia patients and K562 leukemic cells compared with healthy controls and PBMCs. Downregulation of NUDT21 expression in K562 cells inhibits proliferation and promotes apoptosis. Screening by mRNA chip and intracellular signaling array, we found that MAPK/ERK pathway represented the main molecular mechanism underlying the effects of NUDT21 knockdown in K562 cells. CONCLUSION: NUDT21 played an important role in promoting proliferation and inhibiting apoptosis in leukemia K562 cells. The underlying mechanisms involved the modulation of PTEN and a set of downstream molecules including ERK1/2. IMPACT STATEMENT: The present work shows that the expression of NUDT21 was upregulated in chronic myelocytic leukemia and K562 cells. Silencing NUDT21 inhibited the proliferation and promoted the apoptosis of K562 cells. Subsequent experiments confirmed that NUDT21 promoted K562 proliferation through regulating the expression of p-ERK. Our findings may provide insights into the molecular mechanism underlying the effects of NUDT21 on leukemia cells and a novel strategy for the treatment of leukemia.
Authors: Justin Brumbaugh; Bruno Di Stefano; Xiuye Wang; Marti Borkent; Elmira Forouzmand; Katie J Clowers; Fei Ji; Benjamin A Schwarz; Marian Kalocsay; Stephen J Elledge; Yue Chen; Ruslan I Sadreyev; Steven P Gygi; Guang Hu; Yongsheng Shi; Konrad Hochedlinger Journal: Cell Date: 2017-12-14 Impact factor: 41.582
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