Duan Guangchao, Zhang Chuanlin, Xu Changke, Xu Chao, Zhang Lei, Zhang Yan
Department of Spine Surgery, Shangqiu First People's Hospital, Shangqiu, Henan 476100, P.R. China.
Int J Oncol. 2019 Jan;54(1):17-28. doi: 10.3892/ijo.2018.4600. Epub 2018 Oct 19.
Long non‑coding (lnc)RNAs have been demonstrated to be involved in the development of various types of cancers, such as osteosarcoma (OS). Long non‑coding (lnc)RNA metastasis associated lung adenocarcinoma transcript 1 (MALAT1) expression was reported to be highly expressed in OS and promoted the development of this disease; however, the underlying molecular mechanism by which MALAT1 promotes the progression of OS requires further investigation. In the present study, the expression of MALAT1 and miR‑34a was detected by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). The abundance of cyclin D1 (CCND1) was detected by RT‑qPCR and western blotting. Cell viability, migration and invasion were examined by MTT and Transwell assays. The interaction between miR‑34a and MALAT1 or CCND1 was probed by a dual luciferase reporter assay and RNA immunoprecipitation. Xenograft tumor assay was performed to verify the roles of MALAT1 and miR‑34a in tumor growth in vivo. The results demonstrated that MALAT1 and CCND1 mRNA expression levels were upregulated and miR‑34a was downregulated in OS tissues and cells. Additionally, MALAT1 expression was correlated with tumor size, clinical stage and distant metastasis in patients with OS. In addition, MALAT1 promoted OS cell viability, invasion and migration, while MALAT1 silencing exhibited opposing effects. Moreover, MALAT1 functioned as a ceRNA to suppress miR‑34a expression and in turn upregulate CCND1 in OS cells. Rescue experiments further demonstrated that MALAT1 knockdown partially reversed anti‑miR‑34a‑mediated promotion on OS cell viability, migration and invasion; overexpression of CCND1 partially reversed the effects of MALAT1 silencing on OS progression. Furthermore, in vivo experiments also revealed that MALAT1 promoted OS tumor growth via miR‑34a inhibition and upregulating the expression of CCND1. In conclusion, the present study suggested that MALAT1 exerted its oncogenic function in OS by regulating the miR‑34a/CCND1 axis in OS, which may provide novel insight into the diagnosis and therapy for OS.
长链非编码(lnc)RNA已被证明参与多种类型癌症的发生发展,如骨肉瘤(OS)。据报道,长链非编码(lnc)RNA转移相关肺腺癌转录本1(MALAT1)在骨肉瘤中高表达,并促进该疾病的发展;然而,MALAT1促进骨肉瘤进展的潜在分子机制仍需进一步研究。在本研究中,通过逆转录定量聚合酶链反应(RT-qPCR)检测MALAT1和miR-34a的表达。通过RT-qPCR和蛋白质免疫印迹法检测细胞周期蛋白D1(CCND1)的丰度。通过MTT法和Transwell实验检测细胞活力、迁移和侵袭能力。通过双荧光素酶报告基因检测和RNA免疫沉淀实验探究miR-34a与MALAT1或CCND1之间的相互作用。进行异种移植瘤实验以验证MALAT1和miR-34a在体内肿瘤生长中的作用。结果表明,在骨肉瘤组织和细胞中,MALAT1和CCND1 mRNA表达水平上调,而miR-34a表达下调。此外,MALAT1表达与骨肉瘤患者的肿瘤大小、临床分期和远处转移相关。另外,MALAT1促进骨肉瘤细胞的活力、侵袭和迁移,而沉默MALAT1则表现出相反的效果。此外,在骨肉瘤细胞中,MALAT1作为竞争性内源RNA(ceRNA)抑制miR-34a的表达,进而上调CCND1的表达。挽救实验进一步证明,敲低MALAT1可部分逆转抗miR-34a介导的对骨肉瘤细胞活力、迁移和侵袭的促进作用;过表达CCND1可部分逆转沉默MALAT1对骨肉瘤进展的影响。此外,体内实验还表明,MALAT1通过抑制miR-34a并上调CCND1的表达促进骨肉瘤肿瘤生长。总之,本研究表明MALAT1通过调节骨肉瘤中的miR-34a/CCND1轴发挥致癌作用,这可能为骨肉瘤的诊断和治疗提供新的见解。