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Author Spotlight: RNA FISH for Locating lncRNA-SNHG6 in Osteosarcoma Cells
Published on: June 16, 2023
[Long non-coding RNA RAB11B-AS1 prevents osteosarcoma proliferation via its sense gene RAB11B]
1Department of Orthopedics, the Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, China.
Abstract:
Objective: To detect the expression of lncRNA RAB11B-AS1 in osteosarcoma and investigate its role in osteosarcoma cells proliferation and the responsible mechanisms. Methods: Osteosarcoma and corresponding adjacent normal tissues were collected from 24 patients subjected to operations from October 2015 to October 2017 in the Third Affiliated Hospital of Southern Medical University.RAB11B-AS1 expression was detected in osteosarcoma specimens by quantitative real-time polymerase chain reaction (qRT-PCR). Lentiviral vectors that stably over-expressing RAB11B-AS1 were constructed and transfected into U2OS osteosarcoma cell line.The effect of RAB11B-AS1 on osteosarcoma cell proliferation and apoptosis was investigated by cell counting kit (CCK-8) assay and flow cytometry.U2OS osteosarcoma xenograft model of nude mice was established to observe the effect of RAB11B-AS1 on xenograft growth in mice, and the role of RAB11B-AS1 in proliferation and apoptosis of osteosarcoma cells was investigated by immunohistochemistry and TUNEL staining of osteosarcoma slices.The relationship between RAB11B-AS1 and RAB11B was explored using luciferase reporter assay.The data were compared with t test between the two groups. Results: Expression of RAB11B-AS1 was significantly down-regulated in osteosarcoma (0.010±0.015) versus their paired non-neoplastic tissues (0.022±0.030) (t=2.117, P=0.045). Up-regulation of RAB11B-AS1 resulted in decreased proliferative rate of U2OS cells (F=15.659, P<0.001). The ratios of cells in G0-G1 phase, S phase, G2-M phase were 62.6%±6.3%, 21.4%±2.2%, 16.3%±1.6% respectively in RAB11B-AS1 up-regulated group versus 59.4%±5.9%, 25.9%±2.6%, 15.5%±1.1% respectively in control group, and cell ratio in G0-G1 and S phase were increased significantly by RAB11B-AS1 up-regulation (t=17.124, 17.321, both P<0.05). Apoptosis rate was significantly elevated in RAB11B-AS1 over-expressed cells (12.7%±1.3%) when compared with that in control (10.3%±1.0%)(t=17.321, P=0.003). Mice transplanted with osteosarcoma cells that overexpressed RAB11B-AS1 exhibited lower growth rate of tumor (F=8.798, P=0.009). Mechanistically, RAB11B-AS1 expression correlated negatively with RAB11B expression (r=-0.356, P=0.044). Conclusions: lncRNA RAB11B-AS1 expression is down-regulated significantly in osteosarcoma tissues.RAB11B-AS1 may suppress the progression of osteosarcoma via down-regulating RAB11B.
Insights
Long non-coding RNA RAB11B-AS1 (lncRNA RAB11B-AS1) is downregulated in osteosarcoma. Upregulating lncRNA RAB11B-AS1 suppresses osteosarcoma cell proliferation and tumor growth by downregulating RAB11B.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Osteosarcoma is a primary malignant bone tumor with a high mortality rate.
- Long non-coding RNAs (lncRNAs) play crucial roles in various cancers, including osteosarcoma.
- The specific role of lncRNA RAB11B-AS1 in osteosarcoma remains largely unexplored.
Purpose of the Study:
- To investigate the expression levels of lncRNA RAB11B-AS1 in osteosarcoma tissues and cells.
- To explore the functional role of lncRNA RAB11B-AS1 in osteosarcoma cell proliferation, apoptosis, and tumor growth.
- To elucidate the underlying molecular mechanisms by which lncRNA RAB11B-AS1 exerts its effects in osteosarcoma.
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect lncRNA RAB11B-AS1 expression in osteosarcoma and adjacent normal tissues.
- Lentiviral vectors were employed to overexpress lncRNA RAB11B-AS1 in U2OS osteosarcoma cells.
- Cell proliferation was assessed using the cell counting kit (CCK-8) assay, and apoptosis was analyzed by flow cytometry.
- An in vivo osteosarcoma xenograft model in nude mice was established to evaluate the effect of lncRNA RAB11B-AS1 on tumor growth.
- Immunohistochemistry and TUNEL staining were performed on tumor tissues.
- Luciferase reporter assays were utilized to investigate the relationship between lncRNA RAB11B-AS1 and RAB11B.
Main Results:
- lncRNA RAB11B-AS1 expression was significantly downregulated in osteosarcoma tissues compared to adjacent normal tissues (P=0.045).
- Overexpression of lncRNA RAB11B-AS1 significantly inhibited osteosarcoma cell proliferation (P<0.001) and induced apoptosis (P=0.003).
- lncRNA RAB11B-AS1 overexpression led to cell cycle arrest at the G0-G1 phase.
- In vivo studies showed that lncRNA RAB11B-AS1 overexpression suppressed tumor growth in nude mice (P=0.009).
- lncRNA RAB11B-AS1 expression was negatively correlated with RAB11B expression (P=0.044).
Conclusions:
- lncRNA RAB11B-AS1 is significantly downregulated in osteosarcoma.
- lncRNA RAB11B-AS1 functions as a tumor suppressor in osteosarcoma.
- lncRNA RAB11B-AS1 may suppress osteosarcoma progression by downregulating RAB11B.
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