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Published on: August 19, 2025
Comparative proteomics and molecular mechanical analysis in CDA-II induced therapy of LCI-D20 hepatocellular
Hui-zhi Fan1, Hang Liu, Chen Zhang
1Chemistry Department of Fudan University, 220 Handan Rd, 200433, Shanghai, China. hzfan@fudan.edu.cn
Purpose:
To investigate the differential proteins and related molecular mechanism of CDA-II (cell differentiation agent-II) induced therapy on a human hepatocellular carcinoma model in nude mice with high metastatic potential (LCI-D20).
Methods:
After tumors were transplanted 11 days, mice were intraperitoneally injected with CDA-II (1,800 mg/kg) for 20 days continuously. The tumor growth-inhibitory efficiency in CDA-II treated groups was calculated. Proteins extracted from tumor tissue were separated by two-dimensional gel electrophoresis (2DE) and the differential proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Western blotting (WB) was performed to verify the expression of certain candidate proteins. Reverse transcription-polymerase chain reaction (RT-PCR) was engaged to study the molecular mechanism of the therapy.
Results:
CDA-II suppressed the growth and metastasis of tumor. The tumor growth-inhibitory efficiency was 41.8%. In total, 27 differentially expressed proteins were identified, including HSP27, UGDH, CK8, Hsp60, ENOA and AnxA5, with functions involved in oncogene expression and/or cell differentiation. In addition, apparent alternations of HSP60 and beta-actin expression levels and their different posttranslational modifications (PTMs) were investigated. RT-PCR analysis confirmed that the cancer related genes c-myc, N-ras and MMP-9 were significantly down-regulated.
Conclusion:
Our results demonstrate that CDA-II presence can change the proteome profiling and favors of the tumor suppression in LCI-D20 cell differentiation. Our results also suggest that the dynamic PTM of HSP60 expression levels could be used to predict HCC and might be a promising and useful biomarker to prognosticate CDA-II therapeutic efficacy.
Insights
Cell differentiation agent-II (CDA-II) therapy effectively suppressed hepatocellular carcinoma growth and metastasis in mice. CDA-II altered protein expression and may serve as a biomarker for treatment efficacy.
Area of Science:
- Oncology
- Molecular Biology
- Proteomics
Background:
- Hepatocellular carcinoma (HCC) is a significant global health concern.
- High metastatic potential HCC models are crucial for therapeutic development.
- Understanding molecular mechanisms of novel therapies is essential for effective treatment.
Purpose of the Study:
- To investigate the effects of cell differentiation agent-II (CDA-II) on a high-metastatic human HCC model (LCI-D20) in nude mice.
- To identify differential proteins and elucidate the molecular mechanisms underlying CDA-II therapy.
Main Methods:
- Tumor-bearing mice were treated with CDA-II.
- Tumor growth and metastasis were assessed.
- Proteomic analysis using 2D-gel electrophoresis and MALDI-TOF-MS identified differentially expressed proteins.
- Western blotting and RT-PCR were used to validate protein expression and gene regulation.
Main Results:
- CDA-II demonstrated a 41.8% tumor growth-inhibitory efficiency, suppressing tumor growth and metastasis.
- 27 differentially expressed proteins were identified, including HSP27, UGDH, CK8, Hsp60, ENOA, and AnxA5, involved in oncogene expression and cell differentiation.
- Down-regulation of cancer-related genes (c-myc, N-ras, MMP-9) and alterations in HSP60 and beta-actin expression and post-translational modifications (PTMs) were observed.
Conclusions:
- CDA-II alters proteome profiling, promoting tumor suppression and cell differentiation in LCI-D20 HCC.
- Dynamic PTMs of HSP60 expression levels show potential as a predictive biomarker for HCC and prognostic indicator for CDA-II therapeutic efficacy.
