长非编码RNA FBXL19-AS1对急性髓细胞白血病THP-1细胞增殖与侵袭的影响

Effects of long non-coding RNA FBXL19-AS1 on the proliferation and invasion of acute myeloid leukemia THP-1 cells

目的探究长非编码RNA F-box和富含亮氨酸的重复蛋白19反义RNA1 (lnc RNA FBXL19-AS1)在急性髓细胞白血病(acute myeloid leukemia,AML)THP-1细胞中的作用和机制。方法将THP-1细胞随机分为pc-NC组、pc-FBXL19-AS1组以及pc-FBXL19-AS1+miR-339-3p mimic组。pc-NC组转染pc DNA3.0-NC,pc-FBXL19-AS1组转染FBXL19-AS1过表达质粒,pc-FBXL19-AS1+miR-339-3p mimic组共转染FBXL19-AS1过表达质粒以及miR-339-3p模拟物。以细胞计数试剂盒-8(CCK-8)检测THP-1细胞增殖能力,以Transwell实验检测THP-1细胞侵袭能力,以蛋白质印迹法检测SKI原癌基因(SKI)表达。结果 72 h时,pc-NC组和pc-FBXL19-AS1组光密度分别为0.60±0.05和0.78±0.07;pc-NC组和pc-FBXL19-AS1组的侵袭细胞数分别为75.00±6.38和120.67±9.18;pc-NC组、pc-FBXL19-AS1组和pc-FBXL19-AS1+miR-339-3p mimic组中SKI蛋白表达水平分别为1.03±0.10,2.08±0.21和1.68±0.11;以上数据,pc-NC组和pc-FBXL19-AS1组相比,pc-FBXL19-AS1+miR-339-3p mimic组与pc-FBXL19-AS1组相比,差异均有统计学意义(均P <0.05)。此外,双荧光素酶报告基因实验表明FBXL19-AS1可直接靶向miR-339-3p。结论 FBXL19-AS1可促进THP-1细胞的增殖与侵袭,FBXL19-AS1可靶向miR-339-3p上调SKI的表达。

Objective To investigate the role and mechanism of long noncoding RNA F-box and leucine rich repeat protein 19 antisense RNA1(IncRNA FBXL19-AS1) in acute myeloid leukemia(AML) THP-1 cells.Methods THP-1 cells were divided into 3 groups:pc-NC group,pc-FBXL19-AS1 group,and pc-FBXL19-AS1+miR-339-3 p mimic group.pc-NC group was transfected with pcDNA3.0-NC,pc-FBXL19-AS1 group was transfected with FBXL19-AS1 overexpression plasmid,pc-FBXL19-AS1+miR-339-3 p mimic group was co-transfected with FBXL19-AS1 overexpression plasmid and miR-339-3 p mimic.Cell counting kit-8(CCK-8) was performed to detect the proliferation of THP-1 cells.Transwell assay was implemented to detect the invasion of THP-1 cells.Western blot was executed to detect SKI proto-oncogene(SKI) expression.Results At 72 h,the optical density of pc-NC group and pc-FBXL19-AS1 group were 0.60±0.05 and 0.78±0.07,respectively;the number of invasive cells were 75.00±6.38 and 120.67±9.18,respectively;the expression levels of SKI protein in the pc-NC group,pc-FBXL19-AS1 group,and pc-FBXL19-AS1+miR-339-3 p mi group were 1.03±0.10,2.08±0.21 and 1.68±0.11.Compared between pc-NC group and pc-FBXL19-AS1 group,pc-FBXL19-AS1+miR-339-3 p mimic group and pc-FBXL19-AS1 group,the differences of the factors were significant(all P <0.05).In addition,the dual-luciferase reporter gene experiment showed that FBXL19-AS1 can directly target miR-339-3 p.Conclusion FBXL19-AS1 can promote the proliferation and invasion of THP-1 cells,and FBXL19-AS1 can target miR-339-3 p to up-regulate the expression of SKI.