Abstract
PurposeThis study aimed to explore the role of long non-coding RNA (lncRNA) BDNF-AS in the progression of multiple myeloma (MM).MethodsThe expression of BDNF-AS, miR-125a-5p, and miR-125b-5p in MM serum and cell lines were detected by quantitative reverse transcriptase PCR (qRT-PCR). The binding relationships between miR-125a/b-5p and BDNF-AS or Bcl-2 were predicted by Starbase and verified by luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK-8) assay and 5-ethynyl-2′-deoxyuridine (EdU) staining assay. Cell migration was evaluated by wound healing assay. The expression levels of apoptosis-related proteins were evaluated by Western blot analysis. The role of BDNF-AS was also investigated in a xenograft tumor model in vivo.ResultsBDNF-AS was significantly upregulated, while miR-125a-5p and miR-125b-5p were downregulated in MM serum and corresponding cancer cell lines. Knockdown of BDNF-AS effectively inhibited the proliferation and migration of MM.1S and U266 cells, and co-transfection of miR-125a-5p or miR-125b-5p inhibitor and sh-BDNF-AS enhanced cell proliferation and migration compared with that in sh-BDNF-AS group. Knockdown of miR-125a-5p or miR-125b-5p significantly enhanced the proliferation and migration of MM.1S and U266 cells, and co-transfection of sh-Bcl-2 and miR-125a/b-5p inhibitor inhibited cell proliferation compared with that in miR-125a/b-5p inhibitor group. Moreover, knockdown of BDNF-AS increased the expression levels of apoptosis-related proteins (cleaved caspase 3 and cleaved PARP), while knockdown of miR-125a-5p or miR-125b-5p reduced the expression levels of these apoptosis-related proteins compared with knockdown of BDNF-AS. Furthermore, knockdown of BDNF-AS effectively suppressed MM tumor growth in vivo.ConclusionOur findings revealed that knockdown of BDNF-AS inhibited the progression of MM by targeting the miR-125a/b-5p-Bcl-2 axis, indicating that BDNF-AS might serve as a novel drug target for MM.
Highlights
Multiple myeloma (MM) is a hematologic malignancy caused by the excessive proliferation of plasma cells in bone marrows [1]
The results showed that brain-derived neurotrophic factor antisense RNA (BDNF-AS) was significantly upregulated in MM serum (n = 30) compared with that from healthy subjects (n = 30) (p < 0.01, Fig. 1A)
The results showed that BDNF-AS was significantly upregulated (p < 0.05), while the expression of miR-125a5p and miR-125b-5p were markedly downregulated in the 4 MM cell lines (p < 0.05, Fig. 1G-I)
Summary
Multiple myeloma (MM) is a hematologic malignancy caused by the excessive proliferation of plasma cells in bone marrows [1]. Despite increasing understanding of the pathogenic mechanisms of MM and the discovery of new therapeutic targets, MM is still an incurable disease with a low 5-year overall survival rate [4]. The brain-derived neurotrophic factor antisense RNA (BDNF-AS) is a naturally conserved lncRNA (located at chromosome region 11p14.1 and with 2036 bp in length) and exerts essential functions in various human diseases [14,15,16]. Ai et al, revealed that inhibition of BDNF-AS using lentiviral shRNA silencing inhibited MM cell growth and angiogenesis in the bone marrow milieu in vivo [18]. These evidences confirmed the important role of BDNF-AS in MM. Its underlying molecular mechanisms in MM remain unclear, which attracted us to focus on it
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