Effects Of Long Non-coding RNA BOLA3-AS1 On Proliferation And Apoptosis Of Acute Myeloid Leukemia HL60 Cells

Objective: Our previous study showed that there was a significant difference in the expression of long non-coding RNA BOLA3-AS1(Lnc RNA BOLA3-AS1)between CD123+ and CD123-de novo AML patients.This study initially explored the effect of BOLA3-AS1 on the genes related to the proliferation and apoptosis pathways of acute myeloid leukemia cells,and explored the correlation between BOLA3-AS1 and the immunophenotype CD123/IL-3Rα of acute myeloid leukemia stem cells.Methods:1.Leukemic cell lines HL60 and K562 were cultured in vitro,and flow cytometry was used to detect the immunophenotyping of K562 and HL60 cells.Total RNA was extracted from the cell lines,and the expression of CD123/IL-3Rα and Lnc RNA BOLA3-AS1 was detected by q RT-PCR.The candidate cell line(HL60 cells)was determined according to the results of flow cytometry and q RT-PCR.2.Three specific si RNA interference sequences were designed to knock down BOLA3-AS1,and the expression level of BOLA3-AS1 in HL60 cells was knocked down by liposome transfection technology.The transfection effect was observed by fluorescence microscope and the transfection conditions were continuously optimized,and the knockdown effect was detected by RT-q PCR.The si RNA with the best effect was selected for subsequent experiments.3.CCK8 assay was used to detect the cell proliferation of si RNA interference group,transfection control group(si-NC group)and blank control group at 0h,24 h,48h and 72 h after transfection;4.q RT-PCR was used to detect the expression of CD123 and Bax,c-myc and Bcl-2 in HL60 cells after knocking down BOLA3-AS1.Results:1.The results of the test of the test of the flow cell technology indicate that cd123 is expressed in hl60 cells and k562 cells,and q RT-PCR showed that the expression of CD123 and BOLA3-AS1 in HL60 cells was significantly higher than that in K562cells(p<0.01),so HL60 cells were selected as the research subjects of subsequent experiments.2.The BOLA3-AS1 knockdown cell model was successfully established in HL60 cells.CCK8 assay showed that compared with the negative control group and the blank control group,the proliferation of the BOLA3-AS1 knockdown group was significantly inhibited from 48 hours(p<0.01).3.The expression of CD123 in HL60 cells decreased after knocking down BOLA3-AS1(p<0.01).4.After knocking down BOLA3-AS1,the expression of pro-apoptotic gene Bax in HL60 cells was significantly increased(p<0.01),and the expression of anti-apoptotic genes c-myc and Bcl-2 was significantly decreased(p<0.01).Conclusions:1.Silencing BOLA3-AS1 inhibited the growth of HL-60 cells.2.The expression of CD123 in HL-60 cells decreased after silencing BOLA3-AS1,suggesting that CD123 may be a target gene of BOLA3-AS1.3.BOLA3-AS1 may promote the malignant proliferation of HL60 cells by regulating the apoptosis-related genes Bax,c-myc and Bcl-2.