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MicroRNA-129-5p suppresses cell proliferation, migration and invasion via targeting ROCK1 in osteosarcoma
1Department of Orthopedic Surgery, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.
Abstract:
Osteosarcoma (OS) is the most common primary malignancy of the bone in teenagers and accounts for 20‑35% of all malignant primary bone tumors. Increasing evidence shows that microRNAs (miRNAs) are abnormally expressed in several types of human cancer. miRNAs are necessary to maintain the malignant phenotype of cancer cells and can function as either tumor suppressors or oncogenes. The present study aimed to measure the expression of miRNA‑129‑5p (miR‑129‑5p) in OS, determine the effects of miR‑129‑5p on the malignant behaviors of OS cells, and elucidate the molecular mechanism underlying the oncogenesis and progression of OS. The expression levels of miR‑129‑5p in OS tissues and cell lines were measured using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. SAOS‑2 and U2OS cells were then transfected with miR‑129‑5p mimics or miR‑negative control. The effects of miR‑129‑5p on the proliferation, migration and invasion of SAOS‑2 and U2OS cells in vitro were then evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Transwell migration assay and invasion assays, respectively. In addition, bioinformatics analysis, a luciferase reporter assay, and RT‑qPCR and western blot analyses were used to examine whether Rho‑associated protein kinase 1 (ROCK1) was a direct target of miR‑129‑5p. The mRNA expression of ROCK1 in OS tissues was detected using RT‑qPCR analysis, and the biological roles of ROCK1 in OS cells were also evaluated. The results showed that miR‑129‑5p was significantly downregulated in the OS tissues and cell lines. The re‑expression of miR‑129‑5p suppressed the cell proliferation, migration and invasion of OS cells. In addition, ROCK1 was confirmed as a direct target of miR‑129‑5p. The mRNA expression of ROCK1 was high in OS tissues and inversely correlated with the expression of miR‑129‑5p. The downregulation of ROCK1 inhibited the proliferation, migration and invasion of OS cells. These findings suggested that miR‑129‑5p inhibited cell proliferation, migration and invasion in the development of OS via the negative regulation of ROCK1. The miR‑129‑5p/ROCK1 axis may serve as an efficient target in cancer therapy.
Insights
MicroRNA-129-5p (miR-129-5p) is downregulated in osteosarcoma (OS) and suppresses tumor growth. Restoring miR-129-5p inhibits OS cell proliferation, migration, and invasion by targeting ROCK1, offering a potential therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Osteosarcoma (OS) is a primary bone cancer common in adolescents.
- MicroRNAs (miRNAs) play crucial roles in cancer, acting as tumor suppressors or oncogenes.
- Aberrant miRNA expression is linked to various human cancers, including OS.
Purpose of the Study:
- To investigate the expression of microRNA-129-5p (miR-129-5p) in osteosarcoma.
- To determine the impact of miR-129-5p on the malignant behaviors of OS cells.
- To elucidate the molecular mechanism of miR-129-5p in OS oncogenesis and progression, focusing on its potential target, ROCK1.
Main Methods:
- Quantitative reverse transcription polymerase chain reaction (RT-qPCR) to measure miR-129-5p and ROCK1 mRNA expression in OS tissues and cell lines.
- In vitro assays (MTT, Transwell migration, and invasion assays) to assess the effects of miR-129-5p on OS cell proliferation, migration, and invasion.
- Bioinformatics analysis, luciferase reporter assays, and western blot analysis to confirm ROCK1 as a direct target of miR-129-5p.
Main Results:
- miR-129-5p was significantly downregulated in osteosarcoma tissues and cell lines.
- Overexpression of miR-129-5p suppressed the proliferation, migration, and invasion of OS cells.
- Rho-associated protein kinase 1 (ROCK1) was identified as a direct target of miR-129-5p, with its expression inversely correlated with miR-129-5p levels in OS.
- Downregulation of ROCK1 inhibited OS cell proliferation, migration, and invasion.
Conclusions:
- miR-129-5p functions as a tumor suppressor in osteosarcoma.
- The miR-129-5p/ROCK1 axis plays a critical role in OS development by regulating cell proliferation, migration, and invasion.
- Targeting the miR-129-5p/ROCK1 pathway presents a promising therapeutic strategy for osteosarcoma.
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