LncRNA HAND2-AS1 inhibits proliferation and promotes apoptosis of chronic myeloid leukemia cells by sponging with micRNA-1275

摘要

OBJECTIVE: The long noncoding RNA (lnc) HAND2-AS1 is down-regulated and microRNA-1275 (miR-1275) is up-regulated in many types of cancers. However, their expressions and the relationship between HAND2-AS1 and miR-1275 in chronic myeloid leukemia (CML) remain unknown and to be further investigated. PATIENTS AND METHODS: Bone marrow samples from 30 CML patients and 10 healthy donors were collected; HAND2-AS1 and miR-1275 were detected by RT-PCR. The correlation between HAND2-AS1 and miR-1275 was analyzed. After lentiviral (LV) HAND2-AS1 and miR-1275 inhibitors were respectively transfected into KCL22 and K562 cells, the expressions of HAND2-AS1 and miR-1275 were detected by RT-PCR. MTT assay was performed to evaluate cell viability and mRNA and proteins levels of Bcl-2, Caspase-3, MMP-2, MMP-9 were detected by RT-PCR and Western blot (WB). Luciferase reporter assay was performed to explore the binding site of HAND2-AS1 and miR-1275. RESULTS: Results showed that HAND2-AS1 was significantly downregulated than healthy control (p0.05), and HAND2-AS1 expressions on stages of AP and BP were much lower than that of CP (p0.05). The miR-1275 expression was significantly upregulated than healthy control (p0.05), and the expressions in stages of accelerated phase (AP) and blast phase (BP) were much higher than that in the stage of chronic phase (CP) in CML (p0.05). Furthermore, HAND2-AS1 was negatively correlated with miR-1275 in CML, but not in healthy control (p0.05). After lentiviral HAND2-AS1 transfection, the HAND2-AS1 expression was significantly up-regulated while miR-1275 was significantly down-regulated (p0.05). Then, the cell proliferation was inhibited after 72 h. Furthermore, the mRNA and protein levels of Bcl-2, MMP-2 and MMP-9 were significantly down-regulated (p0.05), while the expression of Caspase-3 was significantly up-regulated (p0.05). Luciferase reporter assay showed that HAND2-AS1 was a target gene of miR-1275. And after treating with miR-1275 inhibitor, HAND2-AS1 was significantly upregulated (p0.01) and cell proliferation was inhibited (p0.01). Furthermore, the expressions of Bcl-2, MMP-2, and MMP-9 were significantly decreased, while Caspase-3 was significantly increased (p0.01). CONCLUSIONS: HAND2-AS1 was downregulated and miR-1275 was upregulated in CML, and HAND2-AS1 inhibited proliferation and promoted apoptosis of CML cells by sponging with microRNA-1275, which might be a novel therapeutic target for CML.