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Evaluation of the Anti-Liver Cancer Activity of Protein Fractions Isolated from Adenium obesum Leaf Extract
Ashkan Hajinourmohammadi1, Jamil Zargan2, Hanieh Jafary1
1Department of Biology, SR.C., Islamic Azad University, Tehran, Iran.
Introduction:
Liver cancer is the third leading cause of cancer-related death. Plantderived therapeutics have played a significant role in preventing and treating many diseases, including cancers. The present study investigated the anticancer properties of protein fractions from the green leaf extract of Adenium obesum (A. obesum) in the laboratory.
Methods:
Protein fractions of leaf extract were separated using reversed-phase high-performance liquid chromatography (RP-HPLC). The cytotoxicity of protein fractions was studied by MTT and sulforhodamine B assays. The apoptotic cell death was examined using the alkaline comet assay, and redox-related indicators were assessed using the catalase enzyme activity assay, glutathione content, and nitric oxide release. The RBC hemagglutination test investigated the possible presence of ribosome-inactivating proteins (RIPs) in the most toxic protein fraction, and the LD50 of the protein fraction with the highest anticancer effects was determined. The amino acid sequence of fraction proteins was determined by the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) method.
Results:
The results showed that protein fraction 8 had the highest toxicity in the HepG2 cell line, with an IC50 of 0.16 μg/mL. This fraction induced hemagglutination in red blood cells at concentrations higher than 65 μg/mL. The apoptosis was induced in the HepG2 cells following treatment with the concentrations of 0.08, 0.16, 0.32, and 0.64 μg/mL. Moreover, the redox potential of the treated cells was changed after treatment. The in vivo cytotoxicity investigation of this fraction in mice showed that it is not toxic for animals in concentrations up to 800 μg/kg, indicating its safety potential for pharmaceutical applications. The protein extract in the aforementioned fraction contained two proteins (22 and 53 kD) as determined by electrophoresis and sequencing methods.
Conclusion:
The findings of this investigation demonstrated that the protein content of fraction 8 derived from A. obesum leaf extract possesses anticancer activity in the HepG2 cell line. The two isolated proteins from this fraction are novel and have been reported for the first time. Further investigations should be performed to evaluate the treatment potential in in vitro/vivo conditions.
Insights
Adenium obesum leaf extract protein fractions show potent anticancer activity against liver cancer cells. Fraction 8, containing novel proteins, demonstrated significant cytotoxicity and safety in preliminary studies.
Area of Science:
- Phytochemistry
- Molecular Biology
- Cancer Research
Background:
- Liver cancer is a major global health concern, ranking as the third leading cause of cancer-related mortality.
- Plant-derived compounds are recognized for their therapeutic potential in managing various diseases, including cancer.
- This study explored the anticancer capabilities of protein fractions isolated from Adenium obesum green leaf extract.
Purpose of the Study:
- To investigate the cytotoxic and apoptotic effects of Adenium obesum leaf extract protein fractions on liver cancer cells (HepG2).
- To identify and characterize the specific proteins responsible for the observed anticancer activity.
- To assess the safety profile of the active protein fraction for potential pharmaceutical development.
Main Methods:
- Protein fractions were isolated using reversed-phase high-performance liquid chromatography (RP-HPLC).
- Cytotoxicity was evaluated using MTT and sulforhodamine B assays.
- Apoptosis, redox indicators, hemagglutination, and LD50 were assessed using specific assays.
- Protein identification was performed using MALDI-TOF MS and electrophoresis.
Main Results:
- Protein fraction 8 exhibited significant toxicity against HepG2 cells with an IC50 of 0.16 μg/mL.
- This fraction induced apoptosis in HepG2 cells and altered cellular redox potential.
- In vivo studies in mice indicated no toxicity up to 800 μg/kg, suggesting a favorable safety profile.
- Two novel proteins (22 and 53 kD) were identified in the active fraction.
Conclusions:
- The protein content of Adenium obesum fraction 8 demonstrates significant anticancer activity against HepG2 liver cancer cells.
- Two novel proteins isolated from this fraction represent a new discovery in anticancer therapeutics.
- Further research is warranted to explore the full therapeutic potential of these findings in both in vitro and in vivo models.
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