Long noncoding RNA DANCR knockdown inhibits proliferation, migration and invasion of glioma by regulating miR-135a-5p/BMI1
Long noncoding RNA DANCR knockdown inhibits proliferation, migration and invasion of glioma by regulating miR-135a-5p/BMI1
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DOI：
10.1186/s12935-020-1123-4
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发表时间：
2020-02-18
影响因子：
5.8
通讯作者：
Wang, Xiaoming
中科院分区：
文献类型：
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作者：
Feng, Lei;Lin, Tao;Wang, Xiaoming
背景胶质瘤是世界上最常见、最具侵袭性的原发性脑肿瘤，死亡率高。已鉴定出lncRNA在各种癌症（包括神经胶质瘤）的肿瘤发生中起关键作用。然而，DANCR在胶质瘤进展中的确切机制仍不清楚。方法采用qRT-PCR方法检测DANCR、miR-135 a-5 p和BMI 1在胶质瘤组织和细胞中的表达水平。CCK-8法检测细胞增殖，transwell法检测细胞迁移和侵袭能力。通过在线软件预测miR-135 a-5 p与DANCR或BMI 1的可能结合位点，并通过荧光素酶报告试验和RNA免疫沉淀（RIP）试验进行验证。Western blot检测BMI 1蛋白的表达。建立异种移植瘤模型，研究DANCR在体内胶质瘤进展中的作用。结果DANCR在胶质瘤组织和细胞中表达上调，miR-135 a-5 p表达下调。DANCR的敲低抑制胶质瘤细胞的增殖、迁移和侵袭。此外，miR-135 a-5 p是DANCR的直接靶点，其表达升高可逆转miR-135 a-5 p抑制介导的胶质瘤进展。miR-135 a-5 p可与BMI 1特异性结合，使BMI 1在胶质瘤组织和细胞中的表达明显升高。此外，DANCR作为一种ceRNA，通过海绵状的miR-135 a-5 p调节BMI 1的表达和BMI 1介导的对胶质瘤进展的影响。此外，抑制DANCR通过调节体内miR-135 a-5 p和BMI 1表达来限制肿瘤生长。结论DANCR基因敲减可通过调控miR-135 a-5 p/BMI 1轴抑制胶质瘤细胞的增殖、迁移和侵袭，为胶质瘤的治疗提供了新的途径。
Background Glioma is the most common and aggressive primary brain tumor with high mortality rate around the world. LncRNAs have been identified to play key roles in tumorigenesis in various cancers, including glioma. However, the precise mechanism of DANCR in progression of glioma remains poorly defined. Methods The expression levels of DANCR, miR-135a-5p and BMI1 were measured by qRT-PCR in glioma tissues and cells. Cell proliferation, migration and invasion were detected by CCK-8 assay and transwell assay, respectively. The possible binding sites of miR-135a-5p and DANCR or BMI1 were predicted by online software and verified using luciferase report assay and RNA immunoprecipitation (RIP) assay. Western blot analysis was carried out to detect the protein of BMI1 expression. A xenograft tumor model was established to investigate the functions of DANCR in glioma progression in vivo. Results DANCR was upregulated and miR-135a-5p was downregulated in glioma tissues and cells. Knockdown of DANCR inhibited cell proliferation, migration and invasion in glioma cells. In addition, miR-135a-5p was a direct target of DANCR, and its elevated expression could reverse miR-135a-5p inhibition-mediated progression of glioma. Moreover, miR-135a-5p could specially bind to BMI1, and the expression of BMI1 was obviously elevated in glioma tissues and cells. Furthermore, DANCR acted as a ceRNA to regulate BMI1 expression and BMI1-mediated effects on progression of glioma by sponging miR-135a-5p. Besides, inhibition of DANCR limited tumor growth by regulating miR-135a-5p and BMI1 expression in vivo. Conclusion DANCR knockdown inhibited cell proliferation, migration and invasion in glioma cells through regulating miR-135a-5p/BMI1 axis, providing viable therapeutic avenues for treatment of glioma.