摘要
目的研究长链非编码RNA LINC00854对急性髓系白血病(AML)白细胞细胞系的增殖、周期和凋亡的影响。方法利用在线GEPIA数据库数据分析LINC00854在正常人(n=70)和AML患者(n=173)中的表达情况;RTqPCR检测LINC00854在人骨髓间充质干细胞(hMSC细胞)和AML白细胞细胞系L99-15、CCRF-CEM、HL-60和THP-1中的表达水平。将AML细胞分为空白组、转染对照序列(si-NC)组、转染LINC00854干扰序列(si-LINC00854)组,四唑化合物(MTS)实验和流式细胞实验分别检测干扰LINC00854表达对HL-60和THP-1细胞增殖、周期和凋亡的影响。结果LINC00854在AML中的表达水平显著高于正常人,差异有统计学意义(P<0.05)。AML白细胞系L99-15、CCRF-CEM、HL-60和THP-1中LINC00854表达水平均高于hMSC细胞,差异有统计学意义(P<0.001);但在HL-60和THP-1细胞中LINC00854表达上调更显著,因此,选择这2株细胞进行后续实验。MTS实验结果表明,与转染si-NC组比较,在48和72 h这2个时间点,转染si-LINC00854组的HL-60和THP-1细胞的吸光度值均下降,差异有统计学意义(P<0.001),其中培养72 h后表现最显著,表明干扰LINC00854表达显著抑制HL-60和THP-1细胞增殖。与转染si-NC组比较,转染si-LINC00854组的HL-60和THP-1细胞中G1期的细胞比例显著上升,而G2期的细胞比例显著下降,差异均有统计学意义(P<0.001),表明干扰LINC00854表达显著抑制HL-60和THP-1细胞周期从S期向G2期转化。与转染si-NC组比较,转染si-LINC00854组的HL-60和THP-1细胞中凋亡细胞比例显著增加,差异有统计学意义(P<0.001),表明干扰LINC00854表达促进细胞凋亡。结论LINC00854抑制AML白细胞细胞系的增殖和周期,促进凋亡。
Objective To investigate the effects of LINC00854 on proliferation,cell cycle and apoptosis of leukocyte cell lines and acute myeloid leukemia(AML).Methods The LINC00854 expression in AML was analyzed by online GEPIA database.The expression of LINC00854 in human bone marrow mesenchymal stem cells(hMSC),and L99-15,CCRFCEM,HL-60 and THP-1 cell lines was measured using RT-qPCR.AML cells were divided into blank group,transfected control sequence(si-NC)group and transfected LINC00854 interference sequence(si-LINC00854)group.The effects of LINC00854 interference on cell proliferation,cell cycle and apoptosis of HL-60 and THP-1 cells were assessed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)and flow cytometry,respectively.Results The expression level of LINC00854 in AML was significantly higher than that in normal subjects(P<0.05).The expression levels of LINC00854 in AML leukocyte lines L99-15,CCRF-CEM,HL-60 and THP-1 were higher than that in hMSCs(P<0.001).However,the expression of LINC00854 was up-regulated more significantly in HL-60 and THP-1 cells.Therefore,these two cells were selected for the following experiments.MTS experiment result showed that compared with the transfected si-NC group,the absorbance values of the HL-60 and THP-1 cells in the transfected siLINC00854 group were significantly decreased at 48 and 72 h(P<0.001),especially cultured at 72 h,indicating that interfering with the expression of LINC00854 significantly inhibited the proliferation of HL-60 and THP-1 cells.Compared with the transfected si-NC group,the proportion of G1 phase in transfected si-LINC00854 group were significantly increased in HL-60 and THP-1 cells,while the proportion of G2 phase were significantly decreased(P<0.001),indicating that interfering with the expression of LINC00854 significantly inhibited the transition of HL-60 and THP-1 cell cycle from G1phase to S and G2 phase.Compared with the transfected si-NC group,the proportion of apoptotic cells in transfected the siLINC00854 group was significantly increased in HL-60 and THP-1 cells(P<0.001),indicating that interfering with the expression of LINC00854 promotes cell apoptosis.Conclusion LINC00854 could suppress proliferation,arrest cell cycle,and promote apoptosis in AML.
作者
卜庆婷
潘振宇
孙钟慧
BU Qing-ting;PAN Zhen-yu;SUN Zhong-hui(Department of Genetics,Northwest Women's and Children's Hospital,Xi'an,Shaanxi 710061;Department of Pharmacy,Xi'an Children's Hospital,Xi'an,Shaanxi 710003;Department of Pharmacy,Xi'an No 5 Hospital,Xi'an,Shaanxi 710082,China)
出处
《热带医学杂志》
CAS
2022年第9期1212-1217,1274,共7页
Journal of Tropical Medicine
基金
陕西省自然科学基础研究计划(2020JQ-929)
