Abstract
Background The apoptosis of nucleus pulposus (NP) cells reduces the number of nucleus pulposus cells in intervertebral disc tissue, resulting in intervertebral disc degeneration (IDD). MicroRNAs (miRNAs) play an important regulatory role in abnormal cell proliferation and apoptosis. Methods The miR-27a-3p expressions in degenerative NP tissue and cells were measured via qPCR. The impacts of miR-27a-3p on the proliferation and apoptosis of human NP cells were evaluated by flow cytometry assays, MTT assays, and western blot analyses. In addition, target scan and luciferase reporter assay were applied to confirm that RASSF5 was directly binding to miR-27a-3p. Western blot was applied to assess the relationship between miR-27a-3p, RASSF5 and MST1/LATS1, and RAS/RAC1 signaling pathway. Results MiR-27a-3p was downregulated in degenerative NP tissues and cells by comparison with the control group. MiR-27a-3p overexpression enhanced cell proliferation and suppressed apoptosis of NP cells, while the above factors showed an opposite tendency after in the miR-27a-3p inhibitor group. The western blot experiment similarly suggested mir-27a-3p apparently downregulated apoptosis-related proteins (Bax and caspase-3) and upregulated antiapoptotic proteins (Bcl-2). In addition, RASSF5 was confirmed to be directly regulated by miR-27a-3p using the luciferase reporter assay. Overexpressed RASSF5 could reverse the effects caused by miR-27a-3p mimic. Finally, miR-27a-3p could downregulate RASSF5 and affected the MST1/LATS1 and RAS/RAC1 pathway. Conclusion MiR-27a-3p may target RASSF5 and enhance cell proliferation and imped cell apoptosis of the nucleus pulposus cells via the MST1/LATS1 and RAS/RAC1 pathway, lessening the degeneration of intervertebral discs.
Highlights
Intervertebral disc degeneration (IDD) plays a cardinal role in the progress of low back pain (LBP) [1]
About 70% of people are deeply affected by LBP which is considered as “the mystery of medical care in the 20th century” [2]. e number of IDD patients is increasing, which will lead to tremendous economic and healthy pressure on patients. e apoptosis of nucleus pulposus (NP) cells was reported to be one essential factor of the underlying mechanism of IDD [3]. e degeneration is mainly characterized by abundant NP cell apoptosis
MiR-27a-3p was lower expressed in degenerative NP tissues by comparison with the control group (Figure 1(a))
Summary
Intervertebral disc degeneration (IDD) plays a cardinal role in the progress of low back pain (LBP) [1]. MicroRNA (miRNA) is a noncoding RNA molecule composed of 20 to 25 nucleotides, which is related to the regulation of cell growth, apoptosis, and migration through inducing or inhibiting of the translation of target genes [4]. Many studies have reported RASSF5 was involved in a series of biological activities, especially related to cell proliferation and apoptosis. RASSF5 has been found to be regulated negatively by miR-532-5p to reduce alleviate IDD [9]. Xu et al suggested RASSF5 enhanced cells apoptosis and suppressed proliferation in osteosarcoma [10]. According to Zhou et al.’s [11] study, miR-27a-3p can enhance cell. MiR-27a-3p was reported to enhance cell proliferation in lung cancer through targeting SLC7A11 [12]. The purpose of present research was to study the functional role of miR-27a-3p in IDD and tried to verify it might impede NP cells apoptosis to inhibit IDD progression via targeting RASSF5
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