Inhibition of NEDD8 NEDDylation induced apoptosis in acute myeloid leukemia cells via p53 signaling pathway

Abstract

MLN4924 is a potent and selective small-molecule inhibitor of NEDD8-activating enzyme, which showed anti-tumor effect in several types of malignant tumor types. However, the mechanism of action of MLN4924 in acute myeloid leukemia (AML) requires further investigation. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was conducted to detect the mRNA levels of genes. Gene expression was knocked down by short hairpin RNA (shRNA). Moreover, the protein expression was detected by Western blotting (WB) assay. The proliferation and apoptosis of AML cells were measured by cell counting kit-8 (CCK8) assay and flow cytometry (FCM). In the present study, we observed that the mRNA expression levels of NEDD8, UBA3, UBE2M and RBX1 in AML patients were up-regulated compared with healthy controls, which were correlated with worse overall survival (OS) of patients. Besides, knockdown of UBA3, UBE2M and RBX1 inhibited the NEDDylation of CULs and increased the protein expression of p53 and p21 in MOLM-13 cell line. In AML cells, MLN4924 inhibited cell proliferation, promoted cell apoptosis, and induced cell cycle arrest at the G2/M phase. As revealed by experiments in vivo and in vitro, the NEDDylation of CULs was significantly inhibited and the p53 signaling pathway was activated after MLN4924 treatment. So we concluded that NEDD8, UBA3, UBE2M and RBX1 may serve as the prognostic biomarkers and novel therapeutic targets for AML. Inhibition of the NEDDylation pathway resulted in an anti-leukemia effect by activating the p53 signaling pathway.

Integrated Bioinformatic Analysis to Evaluate Target Genes and Pathways in Chronic Lymphocytic Leukemia

“…A recent study has revealed that CLL has an increased in ubiquitin-like alterations, with numerous proteins in pertinent pathways being implicated [22]. Studies have shown that the prognostic biomarkers for AML are NEDD8, UBE2M and RBX1 [23]. In our study, we discovered that CLL patients had significantly higher expression levels of the genes for the ubiquitin-like proteins NEDD8, UBE2M, RBX1, FBXO21, SKP1, KLHL9, and CAND1.…”

Roles of Nucleolar Factor RCL1 in Itraconazole Resistance of Clinical Candida albicans Under Different Stress Conditions

“…Subsequently, qRT-PCR analysis was conducted using 2×SYBR Green PCR Master Mix (SR1110, Solarbio, Beijing, China) as outlined previously. 19 The transcription level of RCL1 was normalized to ACTB (β-actin), and the relative expression of RCL1 was calculated using the 2 −ΔΔCt method. The primers used in the qRT-PCR assay were as follows: RCL1-Forward-5'-GCCAACAGGATGCGAGGTTA-3', RCL1-Reverse-5'-TCAGCGACTA AGGTGATGCC-3'; ACTB-Forward-5'-TGTTGACCGAAGCTCCAATGA-3', ACTB-Reverse-5'-ACCGGTGGTTCTACC AGAAG-3'.…”

ncRNAs Orchestrate Chemosensitivity Induction by Neddylation Blockades

“…Furthermore, MLN4924 was demonstrated to possess antitumor properties by inhibiting proliferation, migration, and invasion of cancer cells as well as inducing their apoptosis or senescence and autophagy in vitro and in vivo [40][41][42][43][44]. Apoptosis induced by the pharmaceutical inhibition of neddylation using MLN4924 was shown to suppress tumor growth in a variety of cancers including leukemia [45][46][47], urothelial carcinoma [40], colorectal cancer [48], and Ewing sarcoma [49]. In addition, MLN4924 exhibited anti-tumor effects via the suppression of angiogenesis in various cancer cell types including cervical cancer (HeLa), renal cell carcinoma (Caki-2), human urothelial cell carcinoma (BFTC-905), and pharyngeal squamous cell carcinoma (FaDu) cells [50].…”