Expression and clinical significance of lncRNA RBM5-AS1 in acute myeloid leukemia

doi:10.3969/j.issn.1673-8640.2023.01.008

Abstract

Abstract:

Objective To investigate the expression and clinical significance of long non-coding RNA binding motif protein 5-antisense strand 1（lncRNA RBM5-AS1） in acute myeloid leukemia（AML）. Methods Totally，72 AML patients diagnosed and treated in Nanyang Central Hospital from January 2017 to January 2019 were enrolled as AML group，43 AML patients with complete remission（CR） after chemotherapy were regarded as AML-CR group，and 45 patients with iron deficiency anemia were enrolled as anemia group. The level of lncRNA RBM5-AS1 in bone marrow of patients in each group was determined by real-time quantitative polymerase chain reaction（RT-qPCR）. The diagnostic value of lncRNA RBM5-AS1 in AML was analyzed by receiver operating characteristic（ROC） curve. The relationship between the level of lncRNA RBM5-AS1 and the prognosis of AML patients was analyzed by Kaplan-Meier curve. The influencing factors of the prognosis of AML patients were analyzed by Cox regression analysis. Human AML cell line HL-60 was cultured in vitro and randomly classified into control group，sh-NC group and sh-lncRNA RBM5-AS1 group. The level of lncRNA RBM5-AS1 in HL-60 cells was determined by RT-qPCR. CCK-8 and Transwell assay were used to determine the proliferation，invasion and migration of HL-60 cells. The expressions of Wnt/β-catenin pathway related proteins（Wnt5a，β-catenin，c-Myc and Cyclin D1） were determined by western blot. Results The expression level of lncRNA RBM5-AS1 in AML group was higher than those in AML-CR group and anemia group（P<0.05），while there was no statistical significance for lncRNA RBM5-AS1 between AML-CR group and anemia group（P>0.05）. The results of ROC curve analysis showed that the area under curve of lncRNA RBM5-AS1 in the diagnosis of AML was 0.853，with a sensitivity of 87.50% and a specificity of 84.40%. According to the average level of lncRNA RBM5-AS1，AML patients were classified into high expression group（37 cases） and low expression group（35 cases）. There was statistical significance in French-American-British （FAB）type，the proportion of bone marrow blasts and white blood cell counts between the 2 groups（P<0.05）. The 3-year cumulative survival rate of patients in high expression group of lncRNA RBM5-AS1 was lower than that in low expression group（P<0.05）. Multivariate analysis showed that FAB M4-M5 classification and high expression of lncRNA RBM5-AS1 were risk factors affecting the prognosis of AML patients（P<0.05）. Interference with lncRNA RBM5-AS1 expression inhibited the proliferation，invasion and migration，the protein expressions of Wnt5a，β-catenin，c-Myc，Cyclin D1 of HL-60 cells（P<0.05）. Conclusions The level of lncRNA RBM5-AS1 is highly expressed in the bone marrow fluid of AML patients，which may affect cell proliferation，migration and invasion by regulating Wnt/β-catenin pathway，and thus plays a role in the diagnosis and prognostic evaluation of AML.

Key words: Long non-coding RNA binding motif protein 5-antisense strand 1, Acute myeloid leukemia, Prognosis

CLC Number:

R446.7

WANG Ruijuan, LI Chao, DUAN Lijuan, SHANG Miao, YANG Ruyu. Expression and clinical significance of lncRNA RBM5-AS1 in acute myeloid leukemia[J]. Laboratory Medicine, 2023, 38(1): 39-45.

Figures/Tables 10

References 23

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基因名称	正向引物（5'~3'）	反向引物（5'~3'）
lncRNA RBM5-AS1	TGGGAATGGGGAAGAGAAC	TCTGTTCCTGGGTCTGTGTGC
GAPDH	AACGGATTTGGTCGTATTG	GGAAGATGGTGATGGGATT

基因名称	正向引物（5'~3'）	反向引物（5'~3'）
lncRNA RBM5-AS1	TGGGAATGGGGAAGAGAAC	TCTGTTCCTGGGTCTGTGTGC
GAPDH	AACGGATTTGGTCGTATTG	GGAAGATGGTGATGGGATT

组别	例数	lncRNA RBM5-AS1
AML组	72	1.91±0.34^*#
AML-CR组	43	1.13±0.21
贫血组	45	1.04±0.18
F值		185.890
P值		0.000

组别	例数	lncRNA RBM5-AS1
AML组	72	1.91±0.34^*#
AML-CR组	43	1.13±0.21
贫血组	45	1.04±0.18
F值		185.890
P值		0.000

组别	例数	性别		年龄		NCCN分级		FAB分型
组别	例数	男/例	女/例	<45岁/例	≥45岁/例	低、中危/例	高危/例	M1+M2+M3/例	M4+M5/例
高表达组	37	22	15	14	23	14	23	16	21
低表达组	35	19	16	19	16	20	15	27	8
χ²值		0.196		1.96		2.69		8.593
P值		0.658		0.162		0.101		0.003
组别	CR			骨髓原始细胞比例		白细胞计数/（×10⁹/L）	红细胞计数/（×10¹²/L）	血小板计数/（×10⁹/L）	血红蛋白/（g/L）
组别	是/例	否/例		<50%/例	≥50%/例	白细胞计数/（×10⁹/L）	红细胞计数/（×10¹²/L）	血小板计数/（×10⁹/L）	血红蛋白/（g/L）
高表达组	13	24		20	17	16.42±2.38	2.49±0.27	28.35±4.53	43.82±5.69
低表达组	17	18		8	27	10.17±1.84	2.59±0.33	30.51±5.12	46.11±6.43
χ²值	1.336			7.366		12.416	1.411	1.898	1.602
P值	0.248			0.007		0	0.163	0.062	0.114