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Updated: Jul 25, 2025
TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis
Published on: June 8, 2020
Tandem mass tag (TMT) quantitative protein analysis-based proteomics and parallel reaction monitoring (PRM)
Yunyan Liu1, Xiaoyu Zhang1, Xingguang Ren1
1Department of Orthopedic Surgery, Orthopedic Oncology Institute, Tangdu Hospital, Fourth Military Medical University, Xi'an, China.
Abstract:
Osteosarcoma is one of the most common orthopedic malignancies and is characterized by rapid disease progression and a poor prognosis. Currently, research on methods to inhibit osteosarcoma proliferation is still limited. In this study, we found that MST4 levels were significantly increased in osteosarcoma cell lines and tumor tissues compared to normal controls and demonstrated that MST4 is an influential factor in promoting osteosarcoma proliferation both in vivo and in vitro. Proteomic analysis was performed on osteosarcoma cells in the MST4 overexpression and vector expression groups, and 545 significantly differentially expressed proteins were identified and quantified. The candidate differentially expressed protein MRC2 was then identified using parallel reaction monitoring validation. Subsequently, MRC2 expression was silenced with small interfering RNA (siRNA), and we were surprised to find that this alteration affected the cell cycle of MST4-overexpressing osteosarcoma cells, promoted apoptosis and impaired the positive regulation of osteosarcoma growth by MST4. In conclusion, this study identified a novel approach for suppressing osteosarcoma proliferation. Reduction of MRC2 activity inhibits osteosarcoma proliferation in patients with high MST4 expression by altering the cell cycle, which may be valuable for treating osteosarcoma and improving patient prognosis.
Insights
This study reveals that elevated MST4 promotes osteosarcoma growth. Inhibiting the protein MRC2, influenced by MST4, halts cancer cell proliferation by altering the cell cycle and promoting apoptosis, offering a new therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Osteosarcoma is a prevalent orthopedic cancer with limited treatment options.
- Understanding the molecular mechanisms driving osteosarcoma proliferation is crucial for developing new therapies.
Purpose of the Study:
- To investigate the role of MST4 in osteosarcoma proliferation.
- To identify downstream targets of MST4 and explore their function in osteosarcoma growth.
Main Methods:
- Proteomic analysis of osteosarcoma cells with varying MST4 levels.
- Identification and validation of differentially expressed proteins using parallel reaction monitoring.
- Silencing of candidate protein MRC2 using small interfering RNA (siRNA).
Main Results:
- MST4 levels were significantly elevated in osteosarcoma tissues and cell lines, promoting proliferation in vitro and in vivo.
- Proteomic analysis identified 545 differentially expressed proteins, with MRC2 highlighted as a key candidate.
- Silencing MRC2 in MST4-overexpressing osteosarcoma cells altered cell cycle progression, induced apoptosis, and inhibited tumor growth.
Conclusions:
- MST4 is a significant promoter of osteosarcoma proliferation.
- MRC2 is a downstream effector of MST4, mediating its pro-proliferative effects.
- Targeting MRC2 presents a novel therapeutic strategy for osteosarcoma, particularly in patients with high MST4 expression.
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