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Frozen Skin Tissue Block Preparation: A Protocol to Preserve Cutaneous Melanoma Samples from Murine Dorsal Skin
Published on: April 30, 2023
GSTO1 regards as a meritorious regulator in cutaneous malignant melanoma cells
Li-Kun Wang1, Hai-Long Yue2, Xiao-Jing Peng2
1Department of Dermatology, North China University of Science and Technology Affiliated Hospital, China.
Background:
Glutathione S-transferase omega 1 (GSTO1), as a member of the glutathione S-transferase (GST) family genes, has been discovered to be up-regulated in several cancer cell lines which exhibited strong aggressiveness. However, the function of GSTO1 on cutaneous malignant melanoma (CMM) has not been illuminated.
Methods:
Outcome of expression level and prognosis of GSTO1 were obtained from Oncomine and TCGA database. The specific effects of GSTO1 on the characteristics and regulatory mechanism of CMM cells were demonstrated by cell counting kit-8, colony formation, flow cytometry, and transwell assays in vitro. Western blot was employed to analyze the expression of proliferating cell nuclear antigen (PCNA), p53 and epithelial-to-mesenchymal (EMT) related proteins.
Results:
We observed that GSTO1 was up-regulated in CMM samples when compared with the corresponding controls. Moreover, patients in CMM with high expression of GSTO1 were more likely to have a poor prognosis. Through in vitro experiments, silenced GSTO1 resulted in inhibition of CMM cells growth and aggressiveness, increased cell apoptosis, and blocked cell cycle. Finally, the expression of PCNA, p53 and EMT-related proteins were changed due to reduction of GSTO1.
Conclusions:
To sum up, our outcomes exhibited that weakening GSTO1 reduced the proliferation and mobility of CMM cells, increased the apoptosis ability of CMM cells, and arrested cell cycle at G1 phase, which can be achieved by affecting the expression of PCNA, p53 and the EMT process. This discovery provided a new perspective for elucidating the mechanism of CMM, and offered theoretical support for searching clinical therapeutic targets in the future.
Insights
Glutathione S-transferase omega 1 (GSTO1) is elevated in cutaneous malignant melanoma (CMM), correlating with poor prognosis. Silencing GSTO1 inhibits CMM cell growth, migration, and promotes apoptosis, offering potential therapeutic targets.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Glutathione S-transferase omega 1 (GSTO1) is upregulated in aggressive cancer cell lines.
- The role of GSTO1 in cutaneous malignant melanoma (CMM) remains unclear.
Purpose of the Study:
- To investigate the expression and function of GSTO1 in CMM.
- To determine the prognostic value of GSTO1 in CMM patients.
- To explore the impact of GSTO1 on CMM cell characteristics and regulatory mechanisms.
Main Methods:
- Analysis of GSTO1 expression and prognosis using Oncomine and TCGA databases.
- In vitro assays (cell counting, colony formation, flow cytometry, transwell) to assess CMM cell behavior.
- Western blot analysis of PCNA, p53, and EMT-related proteins.
Main Results:
- GSTO1 expression is significantly higher in CMM samples compared to controls.
- High GSTO1 expression is associated with poorer prognosis in CMM patients.
- GSTO1 silencing inhibited CMM cell proliferation and aggressiveness, induced apoptosis, and arrested cell cycle.
- Reduced GSTO1 altered the expression of PCNA, p53, and EMT-related proteins.
Conclusions:
- GSTO1 downregulation reduces CMM cell proliferation, mobility, and enhances apoptosis by affecting PCNA, p53, and EMT pathways.
- GSTO1 inhibition offers a potential therapeutic strategy for CMM.
- This study provides insights into CMM mechanisms and potential clinical targets.
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