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Anti-cancer effects of recombinant arazyme from Serratia Proteomaculans
Ghazaleh Amjadi1, Kazem Parivar, Seyed Fazlollah Mousavi
1Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Purpose:
Colorectal cancer is a lethal and prevalent type of cancer in both men and women worldwide, which can develop resistance to cancer chemotherapy. Developing an effective therapeutic agent is the most promising method for this life-threatening disease. The present study aimed to identify, clone, express and purify the recombinant arazyme (r-arazyme) of Serratia proteomaculans and evaluate the antitumor effect of r-arazyme in vitro.
Methods:
Bacterial strains and cell line, construction of expression vector and preparation of recombinant protein were prepared and then evaluated by western blot, cell culture, cell viability assay, lactate dehydrogenase release assay, cell apoptosis assay, caspase-3 and -9 activation assay, adhesion assay, matrigel invasion assay and reverse transcriptase-polymerase chain reaction (RT-PCR).
Results:
R-arazyme caused a great cytotoxic effect against human colorectal adenocarcinoma (HT29) cells in a dose-dependent manner, without any cytotoxic effect on human embryonic kidney cells 293 (HEK 293). In addition, r-arazyme could induce apoptosis in colorectal cancer cell lines via caspase-3 activation and the elevation of the Bax/Bcl-2 ratio. Further, r-arazyme inhibited cancer cells angiogenesis by significantly reducing the expression of angiogenesis-related genes such as VEGF, VEGFR-1, and VEGFR-2. Furthermore, r-arazyme could prevent invasion and adhesion of cancer cells. In general, the results may support the evidence that r-arazyme is a promising therapeutic candidate against cancer.
Conclusion:
R-arazyme may play an important role in developing effective therapies against colorectal adenocarcinoma in humans, which results in reducing the overall morbidity and mortality related to colorectal cancer.
Insights
Recombinant arazyme (r-arazyme) from Serratia proteomaculans shows significant antitumor effects against colorectal cancer cells in vitro. This enzyme effectively induces apoptosis and inhibits angiogenesis, offering a promising therapeutic candidate for colorectal cancer treatment.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Colorectal cancer is a leading cause of cancer-related deaths globally.
- Chemotherapy resistance is a significant challenge in treating colorectal cancer.
- Novel therapeutic agents are crucial for improving patient outcomes.
Purpose of the Study:
- To identify, clone, express, and purify recombinant arazyme (r-arazyme) from Serratia proteomaculans.
- To evaluate the in vitro antitumor effects of r-arazyme on colorectal cancer cells.
Main Methods:
- Recombinant protein expression and purification.
- In vitro assays including cell viability, apoptosis, invasion, and adhesion.
- Analysis of caspase activation and gene expression (VEGF, VEGFR-1, VEGFR-2).
Main Results:
- R-arazyme demonstrated dose-dependent cytotoxicity against human colorectal adenocarcinoma (HT29) cells.
- R-arazyme induced apoptosis via caspase-3 activation and altered Bax/Bcl-2 ratio.
- R-arazyme inhibited angiogenesis and reduced cancer cell invasion and adhesion.
Conclusions:
- R-arazyme exhibits potent antitumor properties against colorectal cancer cells in vitro.
- R-arazyme represents a promising therapeutic candidate for colorectal cancer treatment.
- Further research may lead to novel therapies reducing colorectal cancer morbidity and mortality.
