LncRNA DANCR调节miR-656/BMPR1A轴对脑胶质瘤细胞恶性行为的影响

doi:10.19723/j.issn.1671-167X.2026.03.023

摘要/Abstract

摘要：

目的: 探讨长链非编码RNA(long non-coding RNA，LncRNA)分化拮抗非蛋白编码RNA(differentiation antagonizing non-protein coding RNA，DANCR)调节微小RNA-656(miR-656)/骨形态发生蛋白受体1A(bone morphogenetic protein receptor type 1A，BMPR1A)轴对脑胶质瘤细胞免疫微环境的影响。方法: 实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction，qRT-PCR)检测脑胶质瘤细胞中DANCR、miR-656、BMPR1A的表达水平。通过转染或共转染至U87细胞，分为沉默(short hairpin, sh)-DANCR组、过表达(plasmid cytome galoirus, pc)DNA 3.1DANCR组、阴性对照(negative control, NC)sh组、pcDNA 3.1组、sh-DANCR+miR-656抑制剂组、sh-DANCR+NC抑制剂组、sh-DANCR+pcDNA 3.1 BMPR1A组、sh-DANCR+pcDNA 3.1组，并以未处理的U87细胞为空白组。CCK8检测U87细胞增殖；Transwell实验检测侵袭与迁移；流式细胞仪检测细胞凋亡；qRT-PCR检测细胞中DANCR、miR-656、BMPR1A mRNA表达；DANCR与miR-656靶向关系通过双荧光素酶验证；BMPR1A及免疫逃逸因子[程序性死亡受体1(programmed death-1，PD-1)和程序性死亡配体1(programmed death-ligand 1, PD-L1)]通过Western blot检测。结果: U87、A172、LN229和U251细胞较NHA细胞中DANCR、BMPR1A mRNA表达显著增加，miR-656表达显著降低(P＜0.05)；与空白组、sh-DANCR组相比，sh-DANCR组U87细胞增殖率，侵袭、迁移数，DANCR、BMPR1A mRNA、BMPR1A、PD-1、PD-L1表达显著降低，凋亡率、miR-656表达显著增加(P＜0.05)；与pcDNA 3.1组相比，pcDNA 3.1 DANCR组U87细胞增殖率，侵袭、迁移数，DANCR、BMPR1A mRNA、BMPR1A、PD-1、PD-L1表达显著增加，凋亡率、miR-656表达显著降低(P＜0.05)；与sh-DANCR+NC抑制剂组相比，sh-DANCR+miR-656抑制剂组U87细胞增殖率，侵袭、迁移数，BMPR1A mRNA、BMPR1A、PD-1、PD-L1表达显著增加，凋亡率、miR-656表达显著降低(P＜0.05)，DANCR表达的差异无统计学意义(P>0.05)；与sh-DANCR+pcDNA 3.1组相比，sh-DANCR+pcDNA 3.1 BMPR1A组U87细胞增殖率，侵袭、迁移数，BMPR1A mRNA、BMPR1A、PD-1、PD-L1表达显著增加，凋亡率显著降低(P＜0.05)，miR-656表达、DANCR表达差异无统计学意义(P>0.05)。DANCR、miR-656具有靶向负调节关系。结论: LncRNA DANCR调节miR-656/BMPR1A轴改善脑胶质瘤细胞免疫微环境，抑制癌细胞恶性行为发展。

关键词: LncRNA DANCR, miR-656/BMPR1A轴, 脑胶质瘤细胞, 免疫微环境, 恶性行为发展

Abstract:

Objective: To investigate the effect of long non-coding RNA (LncRNA) differentiation antagonizing non-protein coding RNA (DANCR) on the immune microenvironment of glioma cells by regulating the miR-656/bone morphogenetic protein receptor type 1A (BMPR1A) axis. Methods: The expression levels of DANCR, miR-656 and BMPR1A in glioma cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The U87 cells were transfected or co-transfected to form the following groups: sh-DANCR (transfected with sh-DANCR), overexpression (pcDNA 3.1) DANCR (transfected with pcDNA 3.1 DANCR), NC sh (transfected with negative control sh), pcDNA 3.1 (transfected with pcDNA 3.1 vector), sh-DANCR + miR-656 inhibitor (co-transfected with sh-DANCR and miR-656 inhibitor), sh-DANCR + NC inhibitor (co-transfected with sh-DANCR and NC inhibitor), sh-DANCR + pcDNA 3.1 BMPR1A (co-transfected with sh-DANCR and pcDNA 3.1 BMPR1A), and sh-DANCR + pcDNA 3.1 (co-transfected with sh-DANCR and pcDNA 3.1 BMPR1A). The untreated U87 cells were used as the blank group. The proliferation of U87 cells was detected by CCK-8; invasion and migration were detected by Transwell assay; apoptosis was detected by flow cytometry; the expression of DANCR, miR-656, and BMPR1A mRNA in cells was detected by qRT-PCR; the targeting relationship between DANCR and miR-656 was verified by dual-luciferase; and BMPR1A and immune escape factors [programmed death receptor 1 (PD-1) and programmed death-ligand 1 (PD-L1)] were detected by Western blot. Results: The mRNA expressions of DANCR and BMPR1A in U87, A172, LN229 and U251 cells were significantly increased, while the expression of miR-656 was significantly decreased compared with those in NHA cells (P < 0.05). Compared with the blank group and sh-DANCR group, the proliferation rate, invasion, migration number, DANCR, BMPR1A mRNA, BMPR1A, PD-1, PD-L1 expression of U87 cells in the sh-DANCR group were obviously reduced, while the apoptosis rate and miR-656 expression were obviously increased (P < 0.05). Compared with the pcDNA 3.1 group, the proliferation rate, invasion, migration number, DANCR, BMPR1A mRNA, BMPR1A, PD-1, and PD-L1 expression of U87 cells in the pcDNA 3.1 DANCR group were obviously increased, while the apoptosis rate and miR-656 expression were obviously reduced (P < 0.05). Compared with the sh-DANCR +NC inhibitor group, the proliferation rate, invasion, migration number, BMPR1A mRNA, BMPR1A, PD-1, and PD-L1 expression of U87 cells in the sh-DANCR+miR-656 inhibitor group were obviously increased, and the apoptosis rate and miR-656 expression were obviously reduced (P < 0.05), while the expression of DANCR was not obvious (P>0.05). Compared with the sh-DANCR+pcDNA 3.1 group, the proliferation rate, invasion, migration number, BMPR1A mRNA, BMPR1A, PD-1, and PD-L1 expression of U87 cells in the sh-DANCR+pcDNA 3.1 BMPR1A group obviously increased, and the apoptosis rate obviously decreased (P < 0.05), while here was no statistically obvious difference in miR-656 expression and DANCR expression (P>0.05). DANCR and miR-656 had a targeted negative regulatory relationship. Conclusion: LncRNA DANCR improves the immune microenvironment of glioma cells and inhibits the malignant behavior development of cancer cells by regulating the miR-656/BMPR1A axis.

Key words: LncRNA DANCR, MiR-656/BMPR1A axis, Glioma cells, Immune microenvironment, Malignant behavior development

中图分类号:

R739.41

王欧洋, 朱鹏磊, 林杰, 吴昊. LncRNA DANCR调节miR-656/BMPR1A轴对脑胶质瘤细胞恶性行为的影响[J]. 北京大学学报（医学版）, 2026, 58(3): 616-623.

Ouyang WANG, Penglei ZHU, Jie LIN, Hao WU. Effect of LncRNA DANCR on the immune microenvironment of glioma cells by regulating the miR-656/BMPR1A axis[J]. Journal of Peking University (Health Sciences), 2026, 58(3): 616-623.

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Primer name	Orientation	Sequence(5′ to 3′)
DANCR	Forward	GCGCCACTATGTAGCGGGTT
DANCR	Reverse	TCAATGGCTTGTGCCTGTAGTT
miR-656	Forward	GTCAGAAAATGGAGTAACCTTA
miR-656	Reverse	GTCAGAAAATGGAGTAACCTTA
BMPR1A	Forward	TAGTTCGCTGAACCAATAAAGG
BMPR1A	Reverse	GTCAGAAAATGGAGTAACCTTA
β-actin	Forward	GCTAATATCTATAATC
β-actin	Reverse	GAGGCTATCTTCATAGAT
U6	Forward	CTCGCTTCGGCAGCACA
U6	Reverse	AACGCTTCACGAATTTGCGT

Group	DANCR	miR-656	BMPR1A mRNA
NHA	0.94±0.10	0.92±0.10	1.02±0.11
LN229	1.51±0.16^*	0.66±0.07^*	1.41±0.15^*
U251	1.55±0.16^*	0.62±0.07^*	1.47±0.15^*
A172	1.62±0.17^*	0.58±0.06^*	1.57±0.16^*
U87	2.03±0.21^*	0.42±0.05^*	2.03±0.21^*

Group	Proliferation rate/%
Blank	94.22±4.51
NC sh	94.11±4.49
sh-DANCR	47.52±4.82^a，b
pcDNA 3.1	94.24±4.49
pcDNA 3.1 DANCR	125.34±6.85^c
sh-DANCR+NC inhibitor	47.67±4.79
sh-DANCR+miR-656 inhibitor	72.14±7.31^d
sh-DANCR+pcDNA 3.1	47.72±4.81
sh-DANCR+pcDNA 3.1 BMPR1A	72.54±7.38^e

Groups	Invasion count	Migration count
Blank	112.46±11.34	102.61±10.47
NC sh	108.94±11.27	105.81±10.62
sh-DANCR	70.88±7.18^{a, b}	51.46±5.24^{a, b}
pcDNA 3.1	111.74±11.20	104.55±10.53
pcDNA 3.1 DANCR	150.54±15.21^c	145.84±14.75^c
sh-DANCR+NC inhibitor	71.44±7.21	52.37±5.31
sh-DANCR+miR-656 inhibitor	108.72±11.04^d	91.48±9.25^d
sh-DANCR+pcDNA 3.1	71.67±7.20	52.53±5.29
sh-DANCR+pcDNA 3.1 BMPR1A	109.34±11.07^e	91.66±9.28^e

Group	Apoptosis rate/%
Blank	12.51±1.28
NC sh	12.63±1.29
sh-DANCR	37.82±3.86^{a, b}
pcDNA 3.1	12.58±1.28
pcDNA 3.1 DANCR	7.67±0.79^c
sh-DANCR+NC inhibitor	37.64±3.79
sh-DANCR+miR-656 inhibitor	21.04±2.18^d
sh-DANCR+pcDNA 3.1	37.74±3.82
sh-DANCR+pcDNA 3.1 BMPR1A	21.17±2.24^e