Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma
Abstract 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-chai...
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| Format: | Article |
| Language: | English |
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BMC
2023-11-01
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| Series: | Biological Procedures Online |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12575-023-00225-y |
| _version_ | 1797416104969109504 |
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| author | Hien Thi My Ong Eda Ates Oh-Seung Kwon Min-Jung Kang |
| author_facet | Hien Thi My Ong Eda Ates Oh-Seung Kwon Min-Jung Kang |
| author_sort | Hien Thi My Ong |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-03-09T05:58:48Z |
| format | Article |
| id | doaj.art-f45c9adcc2ab42f7865f1a7501906fcb |
| institution | Directory Open Access Journal |
| issn | 1480-9222 |
| language | English |
| last_indexed | 2024-03-09T05:58:48Z |
| publishDate | 2023-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Biological Procedures Online |
| spelling | doaj.art-f45c9adcc2ab42f7865f1a7501906fcb2023-12-03T12:11:57ZengBMCBiological Procedures Online1480-92222023-11-0125112110.1186/s12575-023-00225-yScreening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell CarcinomaHien Thi My Ong0Eda Ates1Oh-Seung Kwon2Min-Jung Kang3Center for Advanced Biomolecular Recognition, Korea Institute of Science and TechnologyCenter for Advanced Biomolecular Recognition, Korea Institute of Science and TechnologyDivision of Bio-Medical Science & Technology, KIST School, University of Science and TechnologyCenter for Advanced Biomolecular Recognition, Korea Institute of Science and TechnologyAbstract 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.https://doi.org/10.1186/s12575-023-00225-yCYP11A1-overexpressing kidney cancer cell modelQuantitative analysis of cholesterol and pregnenoloneLC–MS/MSAutophagyFerroptosis |
| spellingShingle | Hien Thi My Ong Eda Ates Oh-Seung Kwon Min-Jung Kang Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma Biological Procedures Online CYP11A1-overexpressing kidney cancer cell model Quantitative analysis of cholesterol and pregnenolone LC–MS/MS Autophagy Ferroptosis |
| title | Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma |
| title_full | Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma |
| title_fullStr | Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma |
| title_full_unstemmed | Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma |
| title_short | Screening of Natural Compounds for CYP11A1 Stimulation Against Cell Renal Cell Carcinoma |
| title_sort | screening of natural compounds for cyp11a1 stimulation against cell renal cell carcinoma |
| topic | CYP11A1-overexpressing kidney cancer cell model Quantitative analysis of cholesterol and pregnenolone LC–MS/MS Autophagy Ferroptosis |
| url | https://doi.org/10.1186/s12575-023-00225-y |
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