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Updated: Mar 27, 2026
Colony Formation Assay Detecting the Proliferative Capacity of LncRNA-knockdown Osteosarcoma Cells
Published on: January 16, 2026
Long non-coding RNA tumor suppressor candidate 7 functions as a tumor suppressor and inhibits proliferation in
Menglin Cong1, Jianmin Li2, Rui Jing3
1Department of Orthopedic Surgery, Shandong University School of Medicine, Jinan, China.
Abstract:
Osteosarcoma is the most common malignant tumor of bone. Recent studies have proven long non-coding RNAs (lncRNAs) play important roles in the tumorigenesis and progression of cancer. However, few lncRNAs have been investigated in osteosarcoma. Here, we reported a novel lncRNA, tumor suppressor candidate 7 (TUSC7), was significantly downregulated in osteosarcoma tissues compared with paired non-tumor tissues and low expression of TUSC7 indicated poor survival (HR = 0.313, 95 % confidence interval (CI) 0.092-0.867) of osteosarcoma patients. Further analysis revealed that loss copy number of TUSC7 was correlated with low expression of TUSC7, and additionally, loss of TUSC7 copy number also indicated poor prognosis (HR = 3.994, 95 % CI 1.147-13.91) of osteosarcoma patients. Two osteosarcoma cell lines, HOS and MG63, were utilized to investigate biological function of TUSC7. Cell counting kit 8 (CCK-8) assay revealed that after silence of TUSC7, cell proliferation ability increased and the colony formation ability also increased. Further results showed that cell cycle was not affected by treatment of si-TUSC7, while the percentage of apoptotic cells decreased. Western blot showed that after silence of TUSC7, the proapoptotic Bcl2 expression was downregulated. Finally, we established xenograft tumor models in nude mice with MG63 cells. Compared with negative control group, silence of TUSC7 significantly promoted tumor growth in vivo. Thus, we demonstrated that TUSC7 could be a potential tumor suppressor in osteosarcoma.
Insights
Tumor suppressor candidate 7 (TUSC7) is downregulated in osteosarcoma, correlating with poor patient survival. Silencing TUSC7 promotes tumor growth and proliferation, identifying it as a potential tumor suppressor in bone cancer.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Osteosarcoma is the most common bone malignancy.
- Long non-coding RNAs (lncRNAs) are implicated in cancer progression.
- Limited research exists on lncRNAs in osteosarcoma.
Purpose of the Study:
- To investigate the role of the novel lncRNA, tumor suppressor candidate 7 (TUSC7), in osteosarcoma.
- To determine the correlation between TUSC7 expression/copy number and patient prognosis.
- To elucidate the functional impact of TUSC7 on osteosarcoma cell behavior and tumor growth.
Main Methods:
- Analysis of TUSC7 expression and copy number in osteosarcoma tissues.
- Correlation analysis between TUSC7 levels and patient survival data.
- In vitro studies using osteosarcoma cell lines (HOS, MG63) with TUSC7 knockdown (si-TUSC7).
- Cell proliferation (CCK-8), colony formation, cell cycle, and apoptosis assays.
- Western blot analysis for apoptosis-related proteins (Bcl2).
- In vivo tumor xenograft models in nude mice.
Main Results:
- TUSC7 was significantly downregulated in osteosarcoma tissues compared to non-tumor tissues.
- Low TUSC7 expression and loss of TUSC7 copy number were associated with poor patient survival and prognosis.
- TUSC7 knockdown increased osteosarcoma cell proliferation and colony formation.
- TUSC7 silencing decreased apoptosis and downregulated proapoptotic Bcl2 expression.
- Silencing TUSC7 significantly promoted tumor growth in vivo xenograft models.
Conclusions:
- TUSC7 functions as a tumor suppressor in osteosarcoma.
- Downregulation and copy number loss of TUSC7 are critical events in osteosarcoma development and progression.
- TUSC7 represents a potential therapeutic target or biomarker for osteosarcoma.
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