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B-AP15 inhibited colon cancer cell proliferation by decreasing CDK6, Cyclin A, Cyclin E, c-Myc, and VEGF gene
Thanet Sophonnithiprasert1, Sasiwan Konjanthet1, Nattanicha Narinnork1
1Biochemistry Unit, Department of Medical Sciences, Faculty of Science, Rangsit University, Pathum Thani, 12000, Thailand.
Abstract:
This study evaluated the screening of novel possible target proteins of B-AP15 (NSC687852) in silico. Through in vitro investigation, the anticancer activity of B-AP15 was examined with respect to gene expression and cell proliferation of SW620 cells (colon cancer cells). Twenty proteins associated with colon cancer were selected to screen possible target proteins of B-AP15 using molecular docking. The binding position of B-AP15 and the top five candidate proteins were investigated using the Discovery Studio 2021 software program. Cytotoxicity, colonization ability, and cell inhibition were evaluated in SW620 cells for 24, 48, and 72 h via MTT assay, colony formation assay, and trypan blue assay. The gene expression was estimated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) at 48 h. B-AP15 showed higher binding to E2F8 (- 10.88 kcal/mol), Ras (- 10.86 kcal/mol), GSK3B (- 10.77 kcal/mol), CDK6 (- 10.75 kcal/mol), and Raf (- 10.12 kcal/mol) than to USP14 (- 7.83 kcal/mol) as a target protein of B-AP15. Interestingly, Ras, GSK3B, and CDK6 shared the binding position between B-AP15 and known inhibitors. The IC50 values of B-AP15 against SW620 cells for 24, 48, and 72 h were 0.89 ± 0.0019 µM, 1.38 ± 0.16 µM, and 1.77 ± 0.0016 µM, respectively. B-AP15 reduced cell viability and the size and number of colonies and increased cell inhibition in a time- and dose-dependent manner. CDK6, Cyclin A, and VEGF expression were inhibited at B-AP15 concentrations of 0.89 and 1.78 µM after treatment for 48 h. Cyclin E and cMyc expression were suppressed only at 1.78 µM. These results suggest that B-AP15 possesses cytotoxicity, reducing the size and number of colonies and increasing cell inhibition via suppression of CDK6, Cyclin A, Cyclin E c-Myc, and VEGF.
Insights
This study identifies potential new targets for the anticancer drug B-AP15 in colon cancer cells. B-AP15 demonstrated significant cytotoxicity and inhibited key cancer-related genes, suggesting its therapeutic potential.
Area of Science:
- Oncology
- Computational Chemistry
- Molecular Biology
Background:
- B-AP15 (NSC687852) is investigated for its anticancer properties.
- Colon cancer remains a significant health concern, necessitating novel therapeutic strategies.
- Understanding drug-target interactions is crucial for developing effective cancer treatments.
Purpose of the Study:
- To screen potential target proteins of B-AP15 using in silico methods.
- To evaluate the in vitro anticancer activity of B-AP15 on SW620 colon cancer cells.
- To investigate the effects of B-AP15 on gene expression and cell proliferation.
Main Methods:
- In silico molecular docking was employed to screen 20 colon cancer-associated proteins for potential B-AP15 targets.
- The binding interactions of B-AP15 with candidate proteins were analyzed using Discovery Studio 2021.
- In vitro assays including MTT, colony formation, and trypan blue were used to assess cytotoxicity, proliferation, and inhibition in SW620 cells.
- Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was utilized to analyze gene expression changes.
Main Results:
- B-AP15 exhibited strong binding affinity to E2F8, Ras, GSK3B, CDK6, and Raf, with binding energies ranging from -10.12 to -10.88 kcal/mol.
- Ras, GSK3B, and CDK6 showed shared binding positions with known inhibitors, indicating potential common mechanisms of action.
- B-AP15 demonstrated significant time- and dose-dependent cytotoxicity, reducing cell viability, colony formation, and increasing cell inhibition.
- The drug suppressed the expression of CDK6, Cyclin A, VEGF, Cyclin E, and c-Myc in a dose-dependent manner.
Conclusions:
- B-AP15 exhibits potent anticancer activity against SW620 colon cancer cells.
- The drug's mechanism involves the suppression of key genes including CDK6, Cyclin A, VEGF, Cyclin E, and c-Myc.
- Identified potential novel targets like Ras, GSK3B, and CDK6 warrant further investigation for B-AP15's therapeutic efficacy.
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