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Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma
Hien Thi My Ong1,2, Eda Ates1,2, Oh-Seung Kwon2,3
1Center for Advanced Biomolecular Recognition, Korea Institute of Science and Technology, Seoul, 02792, Republic of Korea.
Background:
Renal cancer therapies are challenging owing to the extensive spreading of this cancer to other organs and its ability to pose resistance to current medications. Therefore, drugs targeting novel targets are urgently required to overcome these challenges. The cholesterol side-chain cleavage enzyme (CYP11A1) is closely associated with steroidogenesis, and its downregulation is linked to adrenal dysfunction and several types of carcinoma. We previously found that overexpression of CYP11A1 inhibited epithelial-mesenchymal transition and induced G2/M arrest in the kidney cancer Caki-1 cell line. In this context, natural compounds that exhibit potent CYP11A1 stimulation activity can be promising therpaeutic agents for kidney cancer.
Methods:
We screened a panel of 1374 natural compounds in a wound-healing assay using CYP11A1-transfected Caki-1 cells. Of these, 167 promising biologically active compounds that inhibited cancer cell migration by more than 75% were selected, and their half-maximal inhibitory concentrations (IC50) were determined. The IC50 of 159 compounds was determined and 38 compounds with IC50 values less than 50 µM were selected for further analysis. Steroid hormones (cholesterol and pregnenolone) levels in cells treated with the selected compounds were quantitated using LC-MS/MS to determine their effect on CYP11A1 activity. Western blotting for CYP11A1, autophagy signaling proteins, and ferroptosis regulators were performed to ivestigate the mechanisms underlying the action of the selected compounds.
Results:
We screened five promising natural lead compounds that inhibited cancer cell proliferation after three screening steps. The IC50 of these compounds was determined to be between 5.9 and 14.6 μM. These candidate compounds increased the expression of CYP11A1 and suppressed cholesterol levels while increasing pregnenolone levels, which is consistent with the activation of CYP11A1. Our results showed that CYP11A1 activation inhibited the migration of cancer cells, promoted ferroptosis, and triggered autophagy signaling.
Conclusions:
This study indicates that the CYP11A1-overexpressing Caki-1 cell line is useful for screening drugs against kidney cancer. The two selected compounds could be utilized as lead compounds for anticancer drug discovery, and specifically for the development of antirenal cancer medication.
Insights
Researchers screened natural compounds to find new kidney cancer treatments. Five compounds showed promise by activating the cholesterol side-chain cleavage enzyme (CYP11A1), inhibiting cancer cell migration, and promoting cell death pathways.
Area of Science:
- Biochemistry
- Oncology
- Pharmacology
Background:
- Renal cancer presents therapeutic challenges due to metastasis and drug resistance.
- Novel therapeutic targets are crucial for effective kidney cancer treatment.
- Cholesterol side-chain cleavage enzyme (CYP11A1) downregulation is linked to various carcinomas.
Purpose of the Study:
- To screen natural compounds for kidney cancer therapeutic potential.
- To identify compounds that stimulate CYP11A1 activity in kidney cancer cells.
- To investigate the mechanisms of action for promising anti-cancer compounds.
Main Methods:
- Screened 1374 natural compounds using a wound-healing assay with CYP11A1-transfected Caki-1 cells.
- Determined half-maximal inhibitory concentrations (IC50) for selected compounds.
- Quantitated steroid hormone levels and analyzed protein expression (CYP11A1, autophagy, ferroptosis regulators).
Main Results:
- Identified five natural lead compounds with IC50 values between 5.9 and 14.6 µM.
- Confirmed compounds activated CYP11A1, suppressed cholesterol, and increased pregnenolone levels.
- Observed CYP11A1 activation inhibited cancer cell migration, promoted ferroptosis, and triggered autophagy.
Conclusions:
- CYP11A1-overexpressing Caki-1 cell line is a viable model for screening kidney cancer drugs.
- Two selected compounds show potential as lead compounds for anticancer drug discovery.
- These findings support the development of novel anti-renal cancer medications targeting CYP11A1.
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