CRNDE Regulates PIAS2 Expression In Acute Myeloid Leukemia Prognostic Regulation

Background: Acute Myeloid Leukemia(AML)is a malignant neoplastic disease originated from hematopoietic system.It is characterized by exorbitant proliferation of myeloid progenitor cells,resulted in abnormal aggregation of immature precursors and insufficient production of normal mature blood cells.As a malignant disease with strong heterogeneity,it occurs rapidly and has a high degree of malignancy and a poor prognosis.In recent years,although the understanding of AML has been improved and the treatment methods have been improved,AML is still the leading cause of leukemia-related death.Hence,at present,revealing its potential pathogenesis,identifying new biomarkers and improving therapeutic effect are urgent problems to be solved.Long non-coding RNA(lnc RNA)is considered as the "noise" of genomic transcription,and a by-product of RNA polymerase II transcription,which has no biological function.However,recent studies have shown that lnc RNA is involved in many important regulatory processes of gene,such as silencing of X chromosome,genomic imprinting,chromatin modification,transcriptional activation,transcriptional interference,and intranuclear transport,and has been proved to be associated with many diseases.Some studies have shown tha t long non-coding RNA lnc RNA-Colorectal Neoplasia Differentially Expressed(lnc RNA CRNDE)is associated with a variety of malignant tumors and plays a potential role as competitive endogenous RNA(ceRNA).Also some studies have reported that the expression of CRNDE in AML is increased,but its role in AML and the specific molecular mechanism are not clear yet.Purpose: To established a novel integrated competing endogenous network centred on CRNDE using online data analysis and screened the target mRNAs for prognostic significance.To confirm the differential expression of genes in AML to further validate the network and then to detect the effects of CRNDE on biological behaviour.To elucidate the molecular mechanism of the biological role of CRNDE in regulating the expression of PIAS2 by sponging hsa-mi R-148 a.Materials and methods: Part 1: A novel integrated competing endogenous network was established centred on CRNDE using online data analysis and screened the target mRNAs for prognostic significance.Part 2: Real-time PCR and Western blotting were carried out to confirm the differential expression of genes in AML to further validate the network.The relationship between genes expression level and clinicopathological features was analyzed by statistical methods.Part 3: In AML cell lines,expression level of CRNDE was knocked down with si-CRNDE transient transfection,and the effects of CRNDE on the proliferation and apoptosis of AML cell lines were detected.After single transfection of si-CRNDE or hsa-mi R-148 a mimic in AML cell lines and co-transfection of si-CRNDE and hsa-mi-148 a mimic in AML cell lines,the effects of interfering CRNDE on hsa-micro RNA-148 a and PIAS2 were detected by Real-time-PCR.The expression of PIAS2 was detected by Western blot before and after transfection.After co-transfection of PIAS2 wild/mutant plasmid and hsa-mir-148 a mimic in 293 T cells,the inhibitory effect of hsa-mir-148 a on PIAS2 was detected by double Luciferase Report analysis,in order to confirmed that PIAS2 was the downstream target gene of hsa-mir-148 a.All data results above were analyzed by SPSS19.0 and Graph Pad 7.0.All results with P < 0.05 were considered to be statistically.Result: The competing endogenous network centred on CRNDE was established,and accordingly,PIAS2 was screened as an adverse prognostic factor.The overexpression of CRNDE and PIAS2 and the underexpression of hsa-mi R-148a-5p in AML were tested and verified.High sensitivity and specificity were suggested for early discrimination of AML.CRNDE played a subtle role in promoting proliferation and inhibiting apoptosis.CRNDE directly targets mi R-148a-5p to negatively regulate its expression and then positively regulatesthe expression of PIAS2.Conclusion: CRNDE is a risk factor for AML and regulates PIAS2 mRNA by mimicking hsa-mi R-148a-5p with a potential role in AML pathogenesis,which may adjust and control AML progression.