环状RNA hsa_circ0005273在急性髓系白血病中的表达及其调控肿瘤细胞生长和转移机制研究OA

Objective:To investigate the expression of the circular RNA hsa_circ0005273 in acute myeloid leuke-mia(AML)and its regulatory role and mechanism for tumor cell growth and metastasis.Methods:Bone marrow specimens were collected from 20 patients and from 2 healthy bone marrow donors.Normal human bone marrow stromal cell lines and AML cell lines were cultured.RT-qPCR was used to detect the expression of hsa_circ0005273 and miR-143-3p in clinical samples and cell lines.HL-60 cells were divided into control group,si-NC group(trans-fected with si-NC),and si-hsa_circ0005273 group(transfected with si-hsa_circ0005273),and RT-qPCR was used to detect transfection efficiency,and cell proliferation activity,colony formation,cell cycle distribution,apoptosis,and migration and invasion abilities were evaluated using CCK-8 assay,colony formation assay,flow cytometry,Annexin V-FITC/PI double staining,and Transwell chamber assay.The binding sites of hsa_circ0005273 and miR-143-3p were predicted using the StarBase database,and the targeting relationship between the two was verified using a dual-luciferase reporter assay.Results:Compared with the bone marrow of healthy volunteers,the relative expression level of hsa_circ0005273 was upregulated in the bone marrow of 20 AML patients(P<0.05).The relative expression lev-el of hsa_circ0005273 in AML cell lines was higher than that in normal human bone marrow stromal cell lines(all P<0.05),and the upregulation was most significant in HL-60 cells.Compared with the control group and si-NC group,the relative expression level of hsa_circ0005273 in HL-60 cells in the si-hsa_circ0005273 group was signifi-cantly downregulated,indicating successful transfection.In addition,compared with the control group and si-NC group,the proliferation activity and colony formation number of HL-60 cells in the si-hsa_circ0005273 group de-creased,the proportion of cells in the G2/M phase increased,the apoptosis rate increased,and the migration number and invasion number decreased(all P<0.05).The relative expression level of miR-143-3p in the bone marrow of 20 AML patients and various AML cell lines was lower than that in the bone marrow of healthy volunteers and the nor-mal human bone marrow stromal cell lines,and knockdown of hsa_circ0005273 upregulated the relative expression level of miR-143-3p in HL-60 cells(all P<0.05).Prediction revealed that hsa_circ0005273 and miR-143-3p had complementary binding sites.The relative luciferase activity in cells co-transfected with miR-143-3p mimic and hsa_circ0005273 WT was decreased compared with that in cells transfected with NC mimic and hsa_circ0005273 WT(all P<0.05).Conclusion:Hsa_circ0005273 is highly expressed in AML.Knockdown of hsa_circ0005273 inhibits AML cell proliferation,induces G2/M arrest,and promotes apoptosis,while also inhibiting cell migration and inva-sion.This mechanism may be related to its sponging of miR-143-3p.