β-Arrestin1-Associated LNCRNA RP11-135A1.2 Facilitates The Expression Of HES1 Gene In Acute T Cell Lymphoblastic Leukemia Cell

Objective:Our previous study has shown that the signal molecule β-arrestin1, was enriched in pediatric acute lymphoblastic leukemia, and it could promotes the self-renewal of the leukemia-initiating cell-enriched subpopulation in B-lineage acute lymphoblastic leukemia related to DNMT1 activity. However, the effects and mechanism of β-arrestinl regulating T cell acute lymphoblastic leukemia (T-ALL) is unknown. In this study, we utilized Jurkat cells, the acute T lymphobcytic leukemia cell line, to investigate whether long noncoding RNA (lncRNA) genes are the key downstream targets of P-arrestinl and its effect molecules in human T cell acute lymphoblastic leukemia.Methods:We established stable cells Jurkat-siβ1 and Jurkat-β1 by applying lentiviral vectors to knock-down or overexpress P-arrestinl in T-ALL Jurkat cells, and Jurkat cells transfected with non-specific-siRNA was as Jurkat-Ctrl. Cell cycle was examined by flow cytometry in Jurkat-sipβ1, Jurkat-β1 and Jurkat-Ctrl cells. The cell proliferation was identified by CCK-8 assay. Then a genome-wide lncRNA expression study were carried out by using the Human LncRNA Array v3.0. To validate the results of lncRNA array, qRT-PCR and Western blotting were used to further analyze the selected lincRNAs and their nearby genes.Results:The results of flow cytometry and CCK8 assays were shown that β-arrestinl facilitated cell growth and affected cell cycle of T-ALL Jurkat cells. The lncRNA microarray detected a number of lncRNAs and mRNAs that were differentially expressed in Jurkat-siβ1 cells compared with Jurkat-Ctrl, including 15 long intergenic non-coding RNAs (lincRNAs), as well as those were associated with the nearby coding genes. By using Real-time RT-PCR and Western blotting, we verified that β-arrestinl enhanced the transcription level of RP11-135A1.2, and increased the expression of its nearby gene HES1 in both mRNA and protein levels. Further study showed that the expression of HES1 was positively regulated by RP11-135A1.2 in Jurkat cells.Conclusion:In T-ALL Jurkat cells, β-arrestinl enhances the expression of HES1 through promoting the transcription of LincRNA RP11-135A1.2, to regulate its cell cycle.