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Updated: Apr 25, 2026
Colony Formation Assay Detecting the Proliferative Capacity of LncRNA-knockdown Osteosarcoma Cells
Published on: January 16, 2026
HMGB1 promotes cellular proliferation and invasion, suppresses cellular apoptosis in osteosarcoma
Qingbing Meng1, Jie Zhao, Hongbing Liu
1Orthopedics Department, Yancheng City No.1 People's Hospital, 16 Yue-He Road, Yancheng, 224005, Jiangsu Province, People's Republic of China.
Abstract:
Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. Unfortunately, treatment failures are common due to the metastasis and chemoresistance, but the underlying molecular mechanism remains unclear. Accumulating evidence indicated that the deregulation of DNA-binding protein high-mobility group box 1 (HMGB1) was associated with the development of cancer. This study aimed to explore the expression of HMGB1 in osteosarcoma tissues and its correlation to the clinical pathology of osteosarcoma and to discuss the role of HMGB1 in the development of osteosarcoma. The results from RT-PCR and Western blot showed that the expression rate of HMGB1 messenger RNA (mRNA) and the expression of HMGB1 in the osteosarcoma tissues were significantly higher than those in normal bone tissue (p < 0.05), the expression rate of HMGB1 mRNA and the expression of HMGB1 in the carcinoma tissues with positive lung metastasis were significantly higher than those without lung metastasis (p < 0.05), and with increasing Enneking stage, the expression rate of HMGB1 mRNA and the expression of HMGB1 also increased (p < 0.05). In order to explore the role of HMGB1 in osteosarcoma, the expression of HMGB1 in the human osteosarcoma MG-63 cell line was downregulated by the technique of RNA interference. Western blot results showed that the protein expression of HMGB1 was significantly decreased in the MG-63 cells from HMGB1-siRNA transfection group (p < 0.05), which suggested that HMGB1 was successfully downregulated in the MG-63 cells. Then the changes in proliferation, apoptosis, and invasion of MG-63 cells were examined by MTT test, PI staining, annexin V staining, and transwell chamber assay. Results showed that the abilities of proliferation and invasion were suppressed in HMGB1 knockdown MG-63 cells, and the abilities of apoptosis were enhanced in HMGB1 knockdown MG-63 cells. The expression of cyclin D1, MMP-9 was downregulated in HMGB1 knockdown MG-63 cells, and the expression of caspase-3 was upregulated in HMGB1 knockdown MG-63 cells. Taken together, the overexpression of HMGB1 in osteosarcoma might be related to the tumorigenesis, invasion, and metastasis of osteosarcoma, which might be a potential target for the treatment of osteosarcoma.
Insights
High-mobility group box 1 (HMGB1) is overexpressed in osteosarcoma, correlating with advanced stages and metastasis. Reducing HMGB1 suppresses tumor growth and invasion, suggesting it as a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Osteosarcoma is the most common primary bone cancer in children and adolescents.
- Treatment failures in osteosarcoma are often due to metastasis and chemoresistance, with unclear molecular mechanisms.
- Deregulation of High-Mobility Group Box 1 (HMGB1) is implicated in various cancers.
Purpose of the Study:
- To investigate HMGB1 expression in osteosarcoma tissues.
- To correlate HMGB1 expression with clinical pathology.
- To elucidate the role of HMGB1 in osteosarcoma development and progression.
Main Methods:
- RT-PCR and Western blot to assess HMGB1 mRNA and protein expression in tumor and normal tissues.
- Analysis of HMGB1 expression in relation to lung metastasis and Enneking stage.
- RNA interference (siRNA) to downregulate HMGB1 in MG-63 osteosarcoma cells.
- MTT assay, PI staining, annexin V staining, and Transwell assays to evaluate cell proliferation, apoptosis, and invasion.
- Western blot to examine the expression of key proteins (cyclin D1, MMP-9, caspase-3).
Main Results:
- HMGB1 mRNA and protein expression were significantly higher in osteosarcoma tissues compared to normal bone tissue.
- Elevated HMGB1 expression correlated with positive lung metastasis and advanced Enneking stages.
- Downregulation of HMGB1 in MG-63 cells significantly suppressed proliferation and invasion.
- HMGB1 knockdown enhanced apoptosis in osteosarcoma cells.
- HMGB1 downregulation led to decreased expression of cyclin D1 and MMP-9, and increased expression of caspase-3.
Conclusions:
- Overexpression of HMGB1 is associated with tumorigenesis, invasion, and metastasis in osteosarcoma.
- HMGB1 may serve as a potential therapeutic target for osteosarcoma treatment.
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