Effect of Betulinic Acid on GPX4 and SLC7A11 Gene Expression in Renal Cell Carcinoma Cell Line
Yıl 2026,
Cilt: 8 Sayı: 1, 28 - 35, 28.02.2026
Leyla Acar İlvan
,
Sibel Kuraş
,
Merve Nur Ataş
,
Hayriye Arzu Ergen
Öz
Aim: Renal carcinoma is the predominant form of kidney cancer, with its frequency rising in recent years; thus, the advancement of novel therapeutic strategies is crucial due to its resistance to therapy in advanced stages. Currently, investigations into the anticancer effects of natural substances are increasing. Betulinic acid is a lupan-type pentacyclic triterpenoid molecule found in various plants that is notable for its apoptotic and antioxidant properties. This study aimed to examine the effect of betulinic acid on the expression levels of the glutathione peroxidase 4 and solute carrier family 7 member 11 genes related to glutathione metabolism in the renal cell carcinoma cell line CAKI-2 and the healthy cell line MRC-5.
Material and Methods: Expression analysis of GPX4 and SLC7A11 genes was performed by quantitative real time-polymerase chain reaction. The glyceraldehyde 3-phosphate dehydrogenase gene was used as housekeeping gene.
Results: A significant increase in solute carrier family 7 member 11 (p=0.014) and glutathione peroxidase 4 (p=0.049) gene expression levels were observed due to 25µM betulinic acid treatment.
Conclusion: Our findings suggest that betulinic acid may play a role in renal cell carcinoma line, especially through the glutathione mechanism.
Etik Beyan
Çalışmada bir insan ya da hayvan örneği kullanılmadığından etik kurul onayına gerek bulunmamaktadır.
Kaynakça
-
Moch H, Cubilla AL, Humphrey PA, Reuter VE, Ulbright TM. The 2016 WHO classification of tumours of the urinary system and male genital organs-Part A: Renal, penile, and testicular tumours. Eur Urol 2016;70(1):93-105.
-
Capitanio U, Bensalah K, Bex A, et al. Epidemiology of renal cell carcinoma. Eur Urol 2019;75(1):74-84.
-
Linehan WM, Schmidt LS, Crooks DR, et al. The metabolic basis of kidney cancer. Cancer Discov 2019;9(8):1006-1021.
-
Rahib L, Wehner MR, Matrisian LM, Nead KT. Estimated projection of US cancer incidence and death to 2040. JAMA Netw Open 2021;4(4):e214708.
-
Al Azab RS, Al-Zubi MT, Aladaileh MAA, et al. Renal cell carcinoma: an overview of the epidemiology, presentation, histopathological characteristics, and surgical treatment variation between old and new era – a cross-sectional study. Int J Surg Open 2024;62(2):140-143.
-
Jiang W, Li X, Dong S, Zhou W. Betulinic acid in the treatment of tumour diseases: application and research progress. Biomed Pharmacother 2021;142:111990.
-
Adepoju FO, Duru KC, Li E, Kovaleva EG, Tsurkan MV. Pharmacological potential of betulin as a multitarget compound. Biomolecules 2023;13(7):1105.
-
An T, Zha W, Zi J. Biotechnological production of betulinic acid and derivatives and their applications. Appl Microbiol Biotechnol 2020;104(8):3339-3348.
-
Adepoju FO, Duru KC, Li E, Kovaleva EG, Tsurkan MV. Pharmacological potential of betulin as a multitarget compound. Biomolecules 2023;13(7):1105.
-
Mullauer FB, Kessler JH, Medema JP. Betulinic acid, a natural compound with potent anticancer effects. Anticancer Drugs 2010;21(3):215-227.
-
Jiang W, Li X, Dong S, Zhou W. Betulinic acid in the treatment of tumour diseases: application and research progress. Biomed Pharmacother 2021b;142:111990.
-
Gasmi A, Nasreen A, Lenchyk L, et al. An update on glutathione’s biosynthesis, metabolism, functions, and medicinal purposes. Curr Med Chem 2023;31(29):4579-4601.
-
Ingold I, Berndt C, Schmitt S, et al. Selenium utilization by GPX4 is required to prevent hydroperoxide-induced ferroptosis. Cell 2018;172(3):409-422.e21.
-
Li Y, Liu C, Fang B, et al. Ferroptosis, a therapeutic target for cardiovascular diseases, neurodegenerative diseases and cancer. J Transl Med 2024;22(1):1-25.
-
Wei J, Zhu L. The role of glutathione peroxidase 4 in the progression, drug resistance, and targeted therapy of nonsmall cell lung cancer. Oncol Res 2025;33(4):863.
-
Lin W, Wang C, Liu G, et al. SLC7A11/xCT in cancer: biological functions and therapeutic implications. Am J Cancer Res 2020;10(10):3106-3126.
-
Stockwell BR, Friedmann Angeli JP, Bayir H, et al. Ferroptosis: a regulated cell death nexus linking metabolism, redox biology, and disease. Cell 2017;171(2):273-285.
-
Jiang X, Stockwell BR, Conrad M. Ferroptosis: mechanisms, biology, and role in disease. Nat Rev Mol Cell Biol 2021;22:266-282.
-
Huang J, Li Q, Wang H, et al. Betulinic acid inhibits glioma progression by inducing ferroptosis through the PI3K/Akt and NRF2/HO-1 pathways. J Gene Med 2025;27:e70011.
-
Ergen A, Iplık ES, Ertugrul B, Atas MN, Kasarcı G, Cakmakoglu B. Examination of the apoptotic effects of betulinic acid on renal cancer cell lines. Marmara Med J 2020;33(3):113-118.
-
Ren Z, Zhang X, Han J. Expression and prognostic significance of ferroptosis-related proteins SLC7A11 and GPX4 in renal cell carcinoma. Protein Pept Lett 2023;30(10):868-876.
-
Al Azab RS, Al-Zubi MT, Aladaileh MAA, et al. Renal cell carcinoma: an overview of the epidemiology, presentation, histopathological characteristics, and surgical treatment variation between old and new era – a cross-sectional study. Int J Surg Open 2024;62(2):140-143.
-
Ataş MN, Ergen A. Betülinik asit ve antikanser etkileri. Experimed 2020;10(3):128-134.
-
Banerjee S, Banerjee S, Bishayee A, et al. Cellular and molecular mechanisms underlying the potential of betulinic acid in cancer prevention and treatment. Phytomedicine 2024;132:155858.
-
Eder-Czembirek C, Erovic BM, Czembirek C, et al. Betulinic acid: a radiosensitizer in head and neck squamous cell carcinoma cell lines. Strahlenther Onkol 2010;186(3):143-148.
-
Géry A, Dubreule C, André V, et al. Chaga (Inonotus obliquus), a future potential medicinal fungus in oncology? A chemical study and a comparison of the cytotoxicity against human lung adenocarcinoma cells (A549) and human bronchial epithelial cells (BEAS-2B). Integr Cancer Ther 2018;17(3):832-843.
-
Ataş MN, Ertuğrul B, İplik ES, Çakmakoğlu B, Ergen A. Effects of betulinic acid on AKT/mTOR pathway in renal cell carcinoma. Turk J Urol 2022;48(1):58-63.
-
Adeleke GE, Adaramoye OA. Betulinic acid abates N-nitrosodimethylamine-induced changes in lipid metabolism, oxidative stress, and inflammation in the liver and kidney of Wistar rats. J Biochem Mol Toxicol 2021;35(11):e22901.
-
Bhaumik A, Prasad S. Studies on the antitumor potentials of betulinic acid against murine ascites Dalton’s lymphoma. Int J Basic Clin Pharmacol 2016;5(4):1664-1671.
-
Liu JP, Cen SY, Xue Z, et al. A class of disulfide compounds suppresses ferroptosis by stabilizing GPX4. ACS Chem Biol 2022;17(12):3389-3406.
-
Wu Z, Geng Y, Lu X, et al. Chaperone-mediated autophagy is involved in the execution of ferroptosis. Proc Natl Acad Sci U S A 2019;116:2996-3005.
-
Chen L, Wang C, Wang Y, Hong T, Zhang G, Cui X. Functions, roles, and biological processes of ferroptosis-related genes in renal cancer: a pan-renal cancer analysis. Front Oncol 2022;11:697697.
-
Sugezawa K, Morimoto M, Yamamoto M, et al. GPX4 regulates tumor cell proliferation via suppressing ferroptosis and exhibits prognostic significance in gastric cancer. Anticancer Res 2022;42(12):5719-5729.
-
Dodson M, Castro-Portuguez R, Zhang DD. NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis. Redox Biol 2019;23:101107.
-
Chen H, Li R, Zhao F, Luan L, Han T, Li Z. Betulinic acid increases lifespan and stress resistance via insulin/IGF-1 signaling pathway in Caenorhabditis elegans. Front Nutr 2022;9:960239.
-
Lin CH, Lin PP, Lin CY, et al. Decreased mRNA expression for the two subunits of system xc−, SLC3A2 and SLC7A11, in WBC in patients with schizophrenia: evidence in support of the hypo-glutamatergic hypothesis of schizophrenia. J Psychiatr Res 2016;72:58-63.
-
Kong L, Zhu L, Yi X, et al. Betulinic acid alleviates spleen oxidative damage induced by acute intraperitoneal exposure to T-2 toxin by activating Nrf2 and inhibiting MAPK signaling pathways. Antioxidants (Basel) 2021;10(2):158.
Betülinik Asidin Renal Hücreli Karsinom Hücre Hattında GPX4 ve SLC7A11 Gen Ekspresyonu Üzerine Etkisi
Yıl 2026,
Cilt: 8 Sayı: 1, 28 - 35, 28.02.2026
Leyla Acar İlvan
,
Sibel Kuraş
,
Merve Nur Ataş
,
Hayriye Arzu Ergen
Öz
Aim: Renal carcinoma is the predominant form of kidney cancer, with its frequency rising in recent years; thus, the advancement of novel therapeutic strategies is crucial due to its resistance to therapy in advanced stages. Currently, investigations into the anticancer effects of natural substances are increasing. Betulinic acid is a lupan-type pentacyclic triterpenoid molecule found in various plants that is notable for its apoptotic and antioxidant properties. This study aimed to examine the effect of betulinic acid on the expression levels of the glutathione peroxidase 4 and solute carrier family 7 member 11 genes related to glutathione metabolism in the renal cell carcinoma cell line CAKI-2 and the healthy cell line MRC-5.
Material and Methods: Expression analysis of GPX4 and SLC7A11 genes was performed by quantitative real time-polymerase chain reaction. The glyceraldehyde 3-phosphate dehydrogenase gene was used as housekeeping gene.
Results: A significant increase in solute carrier family 7 member 11 (p=0.014) and glutathione peroxidase 4 (p=0.049) gene expression levels were observed due to 25µM betulinic acid treatment.
Conclusion: Our findings suggest that betulinic acid may play a role in renal cell carcinoma line, especially through the glutathione mechanism.
Kaynakça
-
Moch H, Cubilla AL, Humphrey PA, Reuter VE, Ulbright TM. The 2016 WHO classification of tumours of the urinary system and male genital organs-Part A: Renal, penile, and testicular tumours. Eur Urol 2016;70(1):93-105.
-
Capitanio U, Bensalah K, Bex A, et al. Epidemiology of renal cell carcinoma. Eur Urol 2019;75(1):74-84.
-
Linehan WM, Schmidt LS, Crooks DR, et al. The metabolic basis of kidney cancer. Cancer Discov 2019;9(8):1006-1021.
-
Rahib L, Wehner MR, Matrisian LM, Nead KT. Estimated projection of US cancer incidence and death to 2040. JAMA Netw Open 2021;4(4):e214708.
-
Al Azab RS, Al-Zubi MT, Aladaileh MAA, et al. Renal cell carcinoma: an overview of the epidemiology, presentation, histopathological characteristics, and surgical treatment variation between old and new era – a cross-sectional study. Int J Surg Open 2024;62(2):140-143.
-
Jiang W, Li X, Dong S, Zhou W. Betulinic acid in the treatment of tumour diseases: application and research progress. Biomed Pharmacother 2021;142:111990.
-
Adepoju FO, Duru KC, Li E, Kovaleva EG, Tsurkan MV. Pharmacological potential of betulin as a multitarget compound. Biomolecules 2023;13(7):1105.
-
An T, Zha W, Zi J. Biotechnological production of betulinic acid and derivatives and their applications. Appl Microbiol Biotechnol 2020;104(8):3339-3348.
-
Adepoju FO, Duru KC, Li E, Kovaleva EG, Tsurkan MV. Pharmacological potential of betulin as a multitarget compound. Biomolecules 2023;13(7):1105.
-
Mullauer FB, Kessler JH, Medema JP. Betulinic acid, a natural compound with potent anticancer effects. Anticancer Drugs 2010;21(3):215-227.
-
Jiang W, Li X, Dong S, Zhou W. Betulinic acid in the treatment of tumour diseases: application and research progress. Biomed Pharmacother 2021b;142:111990.
-
Gasmi A, Nasreen A, Lenchyk L, et al. An update on glutathione’s biosynthesis, metabolism, functions, and medicinal purposes. Curr Med Chem 2023;31(29):4579-4601.
-
Ingold I, Berndt C, Schmitt S, et al. Selenium utilization by GPX4 is required to prevent hydroperoxide-induced ferroptosis. Cell 2018;172(3):409-422.e21.
-
Li Y, Liu C, Fang B, et al. Ferroptosis, a therapeutic target for cardiovascular diseases, neurodegenerative diseases and cancer. J Transl Med 2024;22(1):1-25.
-
Wei J, Zhu L. The role of glutathione peroxidase 4 in the progression, drug resistance, and targeted therapy of nonsmall cell lung cancer. Oncol Res 2025;33(4):863.
-
Lin W, Wang C, Liu G, et al. SLC7A11/xCT in cancer: biological functions and therapeutic implications. Am J Cancer Res 2020;10(10):3106-3126.
-
Stockwell BR, Friedmann Angeli JP, Bayir H, et al. Ferroptosis: a regulated cell death nexus linking metabolism, redox biology, and disease. Cell 2017;171(2):273-285.
-
Jiang X, Stockwell BR, Conrad M. Ferroptosis: mechanisms, biology, and role in disease. Nat Rev Mol Cell Biol 2021;22:266-282.
-
Huang J, Li Q, Wang H, et al. Betulinic acid inhibits glioma progression by inducing ferroptosis through the PI3K/Akt and NRF2/HO-1 pathways. J Gene Med 2025;27:e70011.
-
Ergen A, Iplık ES, Ertugrul B, Atas MN, Kasarcı G, Cakmakoglu B. Examination of the apoptotic effects of betulinic acid on renal cancer cell lines. Marmara Med J 2020;33(3):113-118.
-
Ren Z, Zhang X, Han J. Expression and prognostic significance of ferroptosis-related proteins SLC7A11 and GPX4 in renal cell carcinoma. Protein Pept Lett 2023;30(10):868-876.
-
Al Azab RS, Al-Zubi MT, Aladaileh MAA, et al. Renal cell carcinoma: an overview of the epidemiology, presentation, histopathological characteristics, and surgical treatment variation between old and new era – a cross-sectional study. Int J Surg Open 2024;62(2):140-143.
-
Ataş MN, Ergen A. Betülinik asit ve antikanser etkileri. Experimed 2020;10(3):128-134.
-
Banerjee S, Banerjee S, Bishayee A, et al. Cellular and molecular mechanisms underlying the potential of betulinic acid in cancer prevention and treatment. Phytomedicine 2024;132:155858.
-
Eder-Czembirek C, Erovic BM, Czembirek C, et al. Betulinic acid: a radiosensitizer in head and neck squamous cell carcinoma cell lines. Strahlenther Onkol 2010;186(3):143-148.
-
Géry A, Dubreule C, André V, et al. Chaga (Inonotus obliquus), a future potential medicinal fungus in oncology? A chemical study and a comparison of the cytotoxicity against human lung adenocarcinoma cells (A549) and human bronchial epithelial cells (BEAS-2B). Integr Cancer Ther 2018;17(3):832-843.
-
Ataş MN, Ertuğrul B, İplik ES, Çakmakoğlu B, Ergen A. Effects of betulinic acid on AKT/mTOR pathway in renal cell carcinoma. Turk J Urol 2022;48(1):58-63.
-
Adeleke GE, Adaramoye OA. Betulinic acid abates N-nitrosodimethylamine-induced changes in lipid metabolism, oxidative stress, and inflammation in the liver and kidney of Wistar rats. J Biochem Mol Toxicol 2021;35(11):e22901.
-
Bhaumik A, Prasad S. Studies on the antitumor potentials of betulinic acid against murine ascites Dalton’s lymphoma. Int J Basic Clin Pharmacol 2016;5(4):1664-1671.
-
Liu JP, Cen SY, Xue Z, et al. A class of disulfide compounds suppresses ferroptosis by stabilizing GPX4. ACS Chem Biol 2022;17(12):3389-3406.
-
Wu Z, Geng Y, Lu X, et al. Chaperone-mediated autophagy is involved in the execution of ferroptosis. Proc Natl Acad Sci U S A 2019;116:2996-3005.
-
Chen L, Wang C, Wang Y, Hong T, Zhang G, Cui X. Functions, roles, and biological processes of ferroptosis-related genes in renal cancer: a pan-renal cancer analysis. Front Oncol 2022;11:697697.
-
Sugezawa K, Morimoto M, Yamamoto M, et al. GPX4 regulates tumor cell proliferation via suppressing ferroptosis and exhibits prognostic significance in gastric cancer. Anticancer Res 2022;42(12):5719-5729.
-
Dodson M, Castro-Portuguez R, Zhang DD. NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis. Redox Biol 2019;23:101107.
-
Chen H, Li R, Zhao F, Luan L, Han T, Li Z. Betulinic acid increases lifespan and stress resistance via insulin/IGF-1 signaling pathway in Caenorhabditis elegans. Front Nutr 2022;9:960239.
-
Lin CH, Lin PP, Lin CY, et al. Decreased mRNA expression for the two subunits of system xc−, SLC3A2 and SLC7A11, in WBC in patients with schizophrenia: evidence in support of the hypo-glutamatergic hypothesis of schizophrenia. J Psychiatr Res 2016;72:58-63.
-
Kong L, Zhu L, Yi X, et al. Betulinic acid alleviates spleen oxidative damage induced by acute intraperitoneal exposure to T-2 toxin by activating Nrf2 and inhibiting MAPK signaling pathways. Antioxidants (Basel) 2021;10(2):158.