The Potential of Ethanol Extract of Pasak Bumi Roots (Eurycoma longifolia Jack) as an Anti-Prostate Cancer In Vitro Against PC-3 Cells
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Keywords:
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Anticancer, Eurycoma longifolia Jack, In Silico, In Vitro, PC-3
Abstract
The prevalence of prostate cancer cases in men is expected to continue increase. In 2040, it is estimated that there will be 2.293.818 new cases and a 1.05% increase in the death rate due to prostate cancer. Eurycoma longifolia Jack roots extract (ELE) has potential as an alternative treatment. This study aims to analyze ELE potential as an anti-prostate cancer agent through in silico assay and in vitro assays on the prostate cancer cell line (PC-3). ELE compounds were docked against Casp-3, Casp-8, HAX-1, p27, and PTEN. In vitro assays on PC-3 cells were used, namely cell viability (WST-8), ROS levels; cell cycle; and cell apoptosis (flow cytometry), PC-3 cell senescence (μ-Galactosidase staining), Casp-3; Casp-8; HAX-1; p27; and PTEN gene expression (qRT-PCR). All proteins target were successfully docked with ELE compounds and presented binding interactions. ELE is known to reduce viability, intracellular ROS levels, live cells, necrosis, and reduce HAX-1 gene expression, and inhibit the cell cycle G0/G1 phase. ELE can also increase inhibition, senescence, late and early apoptosis, and Casp-3, Casp-8, p27, and PTEN gene expression. ELE 100 μg/mL is the most effective concentration. ELE has potential as an anti-prostate cancer agent through apoptosis, cell cycle, and antioxidant pathways
References
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Cheng, L., H. Zhai, J. Du, G. Zhang, and G. Shi (2023). Lobetyolin Inhibits Cell Proliferation and Induces Cell Apoptosis by Downregulating ASCT2 in Gastric Cancer. Cytotechnology, 75(5); 435–448.
Daviu, N., Y. Portilla, M. G. de Cedrón, A. R. de Molina, and D. F. Barber (2024). DMSA-Coated IONPs Trigger Oxidative Stress, Mitochondrial Metabolic Reprogramming and Changes in Mitochondrial Disposition, Hindering Cell Cycle Progression of Cancer Cells. Biomaterials, 304; 122409.
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Jothi, S., T. Parumasivam, and N. Mohtar (2023). Eurycoma Longifolia: An Overview on the Pharmacological Properties for the Treatment of Common Cancer. Journal of Public Health in Africa, 14(s1); 2495.
Kamalidehghan, B., S. Ghafouri-Fard, E. Motevaseli, and F. Ahmadipour (2018). Inhibition of Human Prostate Cancer (PC-3) Cells and Targeting of PC-3-Derived Prostate Cancer Stem Cells with Koenimbin, a Natural Dietary Compound from Murraya Koenigii (L.) Spreng. Drug Design, Development and Therapy, 4; 1119–1133.
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Kimura, T. and S. Egawa (2018). Epidemiology of Prostate Cancer in Asian Countries. International Journal of Urology, 25(6); 524–531.
Lee, Y. R., M. Chen, and P. P. Pandolfi (2018). The Functions and Regulation of the PTEN Tumour Suppressor: New Modes and Prospects. Nature Reviews Molecular Cell Biology, 19(9); 547–562.
Li, D., D. Cao, Y. Cui, Y. Sun, J. Jiang, and X. Cao (2023). The Potential of Epigallocatechin Gallate in the Chemoprevention and Therapy of Hepatocellular Carcinoma. Frontiers in Pharmacology, 14; 1201085.
Li, Z., J. Y. Ruan, F. Sun, J. J. Yan, J. L. Wang, Z. X. Zhang, et al. (2019). Relationship Between Structural Characteristics and Plant Sources Along with Pharmacology Research of Quassinoids. Chemical and Pharmaceutical Bulletin, 67(7); 654–665.
Ligasová, A., I. Frydrych, and K. Koberna (2023). Basic Methods of Cell Cycle Analysis. International Journal of Molecular Sciences, 24(4); 3674.
Liu, Y., P. Chen, D. Qi, and L. Chen (2022). Glaucocalyxin A Inhibits the Malignancies of Gastric Cancer Cells by Downregulating MDM2 and RNF6 via MiR-3658 and the SMG1-UPF mRNA Decay Pathway. Frontiers in Oncology, 12; 871169.
Mohamad, M. and Y. Ismail (2022). A Study on Antioxidant Properties of Eurycoma Longifolia (Tongkat Ali). Journal of Pharmacy Negative Results, 13(S07); 146–149.
Otto, T. and P. Sicinski (2017). Cell Cycle Proteins as Promising Targets in Cancer Therapy. Nature Reviews Cancer, 17(2); 93–115.
Priyandoko, D., W. Widowati, and K. Y. Gunawan (2019). Ethanolic Extract of Moringa’s Leaves (Moringa Oleifera) Induces Senescence on Adenocarcinomic Alveolar Basal Epithelial Cells (A549 Cell-Lines). In Proceedings of the International Conference on Science and Health Technology, page 156–159.
Pu, X., S. J. Storr, Y. Zhang, E. A. Rakha, A. R. Green, I. O. Ellis, et al. (2017). Caspase-3 and Caspase-8 Expression in Breast Cancer: Caspase-3 Is Associated with Survival. Apoptosis, 22; 357–368.
Ramadhani, F. A., M. F. Prastika, N. Fikriyah, Isnaeni, and N. W. Diyah (2024). Molecular Docking of Flavonoids from Extract of Roselle (Hibiscus Sabdariffa L.) Calyx on PBP2a as the Basis for Antibacterial Activity Against Methicillin-Resistant Staphylococcus Aureus. Science and Technology Indonesia, 9(2); 487–493.
Rawla, P. (2019). Epidemiology of Prostate Cancer. World Journal of Oncology, 10(2); 63–89.
Reczek, C. R. and N. S. Chandel (2017). The Two Faces of Reactive Oxygen Species in Cancer. Annual Review of Cancer Biology, 1; 79–98.
Rodríguez-Berriguete, G., N. Torrealba, M. A. Ortega, P. Martínez-Onsurbe, G. Olmedilla, R. Paniagua, et al. (2015). Prognostic Value of Inhibitors of Apoptosis Proteins (IAPs) and Caspases in Prostate Cancer: Caspase-3 Forms and XIAP Predict Biochemical Progression After Radical Prostatectomy. BMC Cancer, 15(1); 1–9.
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Silalahi, M. (2020). The Utilization of Pasak Bumi Plants Eurycoma longifolia j as Traditional Medicine and Its Bioactivity. Epidemiology and Health Innovation, 1(2); 34–40.
Srivastava, N. and A. K. Saxena (2023). Caspase-3 Activators as Anticancer Agents. Current Protein and Peptide Science, 24(10); 783–804.
Sun, Q., L. L. Hu, and Q. Fu (2019). MCT4 Promotes Cell Proliferation and Invasion of Castration-Resistant Prostate Cancer PC-3 Cell Line. EXCLI Journal, 18; 187.
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Tong, K. L., K. L. Chan, S. AbuBakar, B. S. Low, H. Q. Ma, and P. F. Wong (2015). The in Vitro and in Vivo Anti-Cancer Activities of a Standardized Quassinoids Composition from Eurycoma Longifolia on LNCaP Human Prostate Cancer Cells. PLoS ONE, 10(3); 0121752.
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Widowati, W., D. K. Jasaputra, P. Onggowidjaja, S. B. Sumitro, M. A. Widodo, and E. Afifah (2019). Effects of Conditioned Medium of Co-Culture IL-2 Induced NK Cells and Human Wharton’s Jelly Mesenchymal Stem Cells (hWJMSCs) on Apoptotic Gene Expression in a Breast Cancer Cell Line (MCF-7). Journal of Mathematics and Fundamental Sciences, 51(3); 205–224.
Widowati, W., D. Priyandoko, L. Lenny, R. Revika, S. Novianti, and H. S. Kusuma (2024). Camellia Sinensis L. Extract Suppresses Inflammation on Acute Respiratory Distress Syndrome Cell Models via Decreasing IL-1ß, IL-6, and COX-2 Expressions. Trends in Sciences, 21(1); 7010–7010.
Widowati, W., T. L. Wargasetia, F. Rahardja, R. F. Gunanegara, D. Priyandoko, and M. E. S. Gondokesumo (2023). hWJMSCs Inhibit Inflammation and Apoptosis in an ARDS Cell Model. Journal of Taibah University Medical Sciences, 18(6); 1519–1526.
Xie, J., Z. Lai, X. Zheng, H. Liao, Y. Xian, and Q. Li (2021). Apoptotic Activities of Brusatol in Human Non-Small Cell Lung Cancer Cells: Involvement of ROS-Mediated Mitochondrial-Dependent Pathway and Inhibition of Nrf2-Mediated Antioxidant Response. Toxicology, 451; 152680.
Yan, J., C. Ma, J. Cheng, Z. Li, and C. Liu (2015). HAX-1 Inhibits Apoptosis in Prostate Cancer Through the Suppression of Caspase-9 Activation. Oncology Reports, 34(5); 2776–2781.
Ye, G., M. Xu, Y. Shu, X. Sun, Y. Mai, and Y. Hong (2022). A Quassinoid Diterpenoid Eurycomanone From Eurycoma Longifolia Jack Exerts Anti-Cancer Effect Through Autophagy Inhibition. Molecules, 27(14); 4398.
Yunos, N. M., N. D. Amin, M. H. Jauri, S. K. Ling, N. H. Hassan, and N. J. Sallehudin (2022). The in Vitro Anti-Cancer Activities and Mechanisms of Action of 9-Methoxycanthin-6-One From Eurycoma Longifolia in Selected Cancer Cell Lines. Molecules, 27(3); 585.
Yunos, N. M., H. A. Wahab, M. G. Al-Thiabat, N. J. Sallehudin, and M. H. Jauri (2023). In Vitro and in Silico Analysis of the Anti-Cancer Effects of Eurycomanone and Eurycomalactone From Eurycoma Longifolia. Plants, 12; 2827.
Zeng, S., W. H. Shen, and L. Liu (2018). Senescence and Cancer. Cancer Translational Medicine, 4(3); 70–74.
Zhong, W., S. Dong, H. Wang, C. Pan, and S. Yang (2023). Functional Mechanism of MicroRNA-25-3p in Hilar Cholangiocarcinoma Cell Proliferation and Migration Through Regulation of Dual Specificity Phosphatase 5. Journal of Investigative Surgery, 36(1); 2202768.
Al-Salahi, O. S., D. Ji, A. M. Majid, C. Kit-Lam, W. Z. Abdullah, A. Zaki, et al. (2014). Anti-Tumor Activity of Eurycoma Longifolia Root Extracts Against K-562 Cell Line: In Vitro and In Vivo Study. PLOS ONE, 9(1); e83818.
Anwar, K., A. Lutpi, A. Melinda, and S. Hadi (2022). Extraction Methods Effect on Antioxidant Activity of Ethanol Extract of ‘Pasak Bumi’ (Eurycoma Longifolia Jack.) Root. In IOP Conference Series: Earth and Environmental Science, volume 976, page 012052.
Bencivenga, D., E. Stampone, A. Aulitto, A. Tramontano, C. Barone, and A. Negri (2021). A Cancer-Associated CDKN1B Mutation Induces p27 Phosphorylation on a Novel Residue: A New Mechanism for Tumor Suppressor Loss-of-Function. Molecular Oncology, 15(4); 915–941.
Bray, F., J. Ferlay, I. Soerjomataram, R. L. Siegel, L. A. Torre, and A. Jemal (2018). Global Cancer Statistics 2018: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA: A Cancer Journal for Clinicians, 68(6); 394–424.
Chen, J., W. G. Nelson, K. Sfanos, S. Yegnasubramanian, and A. M. De Marzo (2023). Molecular Pathology of Prostate Cancer. In Molecular Surgical Pathology 2023, page 377–399.
Cheng, L., H. Zhai, J. Du, G. Zhang, and G. Shi (2023). Lobetyolin Inhibits Cell Proliferation and Induces Cell Apoptosis by Downregulating ASCT2 in Gastric Cancer. Cytotechnology, 75(5); 435–448.
Daviu, N., Y. Portilla, M. G. de Cedrón, A. R. de Molina, and D. F. Barber (2024). DMSA-Coated IONPs Trigger Oxidative Stress, Mitochondrial Metabolic Reprogramming and Changes in Mitochondrial Disposition, Hindering Cell Cycle Progression of Cancer Cells. Biomaterials, 304; 122409.
Druzhilovskiy, D. S., A. V. Rudik, D. A. Filimonov, T. A. Gloriozova, A. A. Lagunin, A. V. Dmitriev, P. V. Pogodin, V. I. Dubovskaya, S. M. Ivanov, O. A. Tarasova, et al. (2017). Computational Platform Way2Drug: From the Prediction of Biological Activity to Drug Repurposing. Russian Chemical Bulletin, 66; 1832–1841.
Ferlay, J., M. Colombet, I. Soerjomataram, C. Mathers, D. M. Parkin, M. Piñeros, et al. (2018). Global Cancer Observatory: Cancer Today. International Agency for Research on Cancer, Lyon, France.
Foster, K., O. Oyenihi, S. Rademan, J. Erhabor, M. Matsabisa, J. Barker, et al. (2020). Selective Cytotoxic and Anti-Metastatic Activity in DU-145 Prostate Cancer Cells Induced by Annona Muricata L. Bark Extract and Phytochemical, Annonacin. BMC Complementary Medicine and Therapies, 20(1).
Ghosh, S., J. Hazra, K. Pal, V. K. Nelson, and M. Pal (2021). Prostate Cancer: Therapeutic Prospect with Herbal Medicine. Current Research in Pharmacology and Drug Discovery, 2; 100034.
Gondokesumo, M. E., A. Novilla, S. Prahastuti, H. S. W. Kusuma, W. Widowati, F. H. Zahiroh, D. S. Hadiprasetyo, and W. Surakusumah (2025). Anticancer Effectivity of Nanocrystals Derived from Mangosteen (Garcinia Mangostana) Peel Extract on Leukemia HL-60 Cells. Science and Technology Indonesia, 10(1); 228–237.
Icard, P., L. Fournel, Z. Wu, M. Alifano, and H. Lincet (2019). Interconnection Between Metabolism and Cell Cycle in Cancer. Trends in Biochemical Sciences, 44(6); 490–501.
Jothi, S., T. Parumasivam, and N. Mohtar (2023). Eurycoma Longifolia: An Overview on the Pharmacological Properties for the Treatment of Common Cancer. Journal of Public Health in Africa, 14(s1); 2495.
Kamalidehghan, B., S. Ghafouri-Fard, E. Motevaseli, and F. Ahmadipour (2018). Inhibition of Human Prostate Cancer (PC-3) Cells and Targeting of PC-3-Derived Prostate Cancer Stem Cells with Koenimbin, a Natural Dietary Compound from Murraya Koenigii (L.) Spreng. Drug Design, Development and Therapy, 4; 1119–1133.
Khanam, Z., C. S. Wen, and I. U. Bhat (2015). Phytochemical Screening and Antimicrobial Activity of Root and Stem Extracts of Wild Eurycoma Longifolia Jack (Tongkat Ali). Journal of King Saud University - Science, 27(1); 23–30.
Kimura, T. and S. Egawa (2018). Epidemiology of Prostate Cancer in Asian Countries. International Journal of Urology, 25(6); 524–531.
Lee, Y. R., M. Chen, and P. P. Pandolfi (2018). The Functions and Regulation of the PTEN Tumour Suppressor: New Modes and Prospects. Nature Reviews Molecular Cell Biology, 19(9); 547–562.
Li, D., D. Cao, Y. Cui, Y. Sun, J. Jiang, and X. Cao (2023). The Potential of Epigallocatechin Gallate in the Chemoprevention and Therapy of Hepatocellular Carcinoma. Frontiers in Pharmacology, 14; 1201085.
Li, Z., J. Y. Ruan, F. Sun, J. J. Yan, J. L. Wang, Z. X. Zhang, et al. (2019). Relationship Between Structural Characteristics and Plant Sources Along with Pharmacology Research of Quassinoids. Chemical and Pharmaceutical Bulletin, 67(7); 654–665.
Ligasová, A., I. Frydrych, and K. Koberna (2023). Basic Methods of Cell Cycle Analysis. International Journal of Molecular Sciences, 24(4); 3674.
Liu, Y., P. Chen, D. Qi, and L. Chen (2022). Glaucocalyxin A Inhibits the Malignancies of Gastric Cancer Cells by Downregulating MDM2 and RNF6 via MiR-3658 and the SMG1-UPF mRNA Decay Pathway. Frontiers in Oncology, 12; 871169.
Mohamad, M. and Y. Ismail (2022). A Study on Antioxidant Properties of Eurycoma Longifolia (Tongkat Ali). Journal of Pharmacy Negative Results, 13(S07); 146–149.
Otto, T. and P. Sicinski (2017). Cell Cycle Proteins as Promising Targets in Cancer Therapy. Nature Reviews Cancer, 17(2); 93–115.
Priyandoko, D., W. Widowati, and K. Y. Gunawan (2019). Ethanolic Extract of Moringa’s Leaves (Moringa Oleifera) Induces Senescence on Adenocarcinomic Alveolar Basal Epithelial Cells (A549 Cell-Lines). In Proceedings of the International Conference on Science and Health Technology, page 156–159.
Pu, X., S. J. Storr, Y. Zhang, E. A. Rakha, A. R. Green, I. O. Ellis, et al. (2017). Caspase-3 and Caspase-8 Expression in Breast Cancer: Caspase-3 Is Associated with Survival. Apoptosis, 22; 357–368.
Ramadhani, F. A., M. F. Prastika, N. Fikriyah, Isnaeni, and N. W. Diyah (2024). Molecular Docking of Flavonoids from Extract of Roselle (Hibiscus Sabdariffa L.) Calyx on PBP2a as the Basis for Antibacterial Activity Against Methicillin-Resistant Staphylococcus Aureus. Science and Technology Indonesia, 9(2); 487–493.
Rawla, P. (2019). Epidemiology of Prostate Cancer. World Journal of Oncology, 10(2); 63–89.
Reczek, C. R. and N. S. Chandel (2017). The Two Faces of Reactive Oxygen Species in Cancer. Annual Review of Cancer Biology, 1; 79–98.
Rodríguez-Berriguete, G., N. Torrealba, M. A. Ortega, P. Martínez-Onsurbe, G. Olmedilla, R. Paniagua, et al. (2015). Prognostic Value of Inhibitors of Apoptosis Proteins (IAPs) and Caspases in Prostate Cancer: Caspase-3 Forms and XIAP Predict Biochemical Progression After Radical Prostatectomy. BMC Cancer, 15(1); 1–9.
Ruan, J., Z. Li, Y. Zhang, Y. Chen, M. Liu, L. Han, Y. Zhang, and T. Wang (2019). Bioactive Constituents from the Roots of Eurycoma Longifolia. Molecules, 24(17); 3157.
Ryu, S. M., J. Kwon, Y. H. Seo, E. G. Song, S. S. Hong, B. S. Kim, J. S. Hong, K. H. Ryu, and D. Lee (2017). Quassinoids Isolated from Brucea Javanica Inhibit Pepper Mottle Virus in Pepper. Virus Research, 227; 49–56.
Silalahi, M. (2020). The Utilization of Pasak Bumi Plants Eurycoma longifolia j as Traditional Medicine and Its Bioactivity. Epidemiology and Health Innovation, 1(2); 34–40.
Srivastava, N. and A. K. Saxena (2023). Caspase-3 Activators as Anticancer Agents. Current Protein and Peptide Science, 24(10); 783–804.
Sun, Q., L. L. Hu, and Q. Fu (2019). MCT4 Promotes Cell Proliferation and Invasion of Castration-Resistant Prostate Cancer PC-3 Cell Line. EXCLI Journal, 18; 187.
Tamizhazhagan, V., K. Pugazhendy, V. Sakthidasan, C. Jayanthi, S. Rajesh, and P. Manikanadan (2017). Social and Economic Burden of Cancer on 2020-Minireview. Journal of Biological and Medical Sciences, 1(103); 2.
Tong, K. L., K. L. Chan, S. AbuBakar, B. S. Low, H. Q. Ma, and P. F. Wong (2015). The in Vitro and in Vivo Anti-Cancer Activities of a Standardized Quassinoids Composition from Eurycoma Longifolia on LNCaP Human Prostate Cancer Cells. PLoS ONE, 10(3); 0121752.
Trebinska-Stryjewska, A., L. Szafron, A. Rembiszewska, M. Wakula, S. Tabor, R. Sienkiewicz, J. Owczarek, A. Balcerak, A. Felisiak-Golabek, and E. A. Grzybowska (2019). Cytoplasmic HAX1 Is an Independent Risk Factor for Breast Cancer Metastasis. Journal of Oncology, 2019.
Widowati, W., R. F. Gunanegara, T. L. Wargasetia, H. S. Kusuma, S. Arumwardana, and C. D. Wahyuni (2021). Effect of Flavonoids on Oxidative Stress, Apoptosis, and Cell Markers of Peripheral Blood-Derived Endothelial Progenitor Cells: An in Vitro Study. International Journal of Applied Pharmaceutics, 13(Special Issue 3); 39–42.
Widowati, W., D. K. Jasaputra, P. Onggowidjaja, S. B. Sumitro, M. A. Widodo, and E. Afifah (2019). Effects of Conditioned Medium of Co-Culture IL-2 Induced NK Cells and Human Wharton’s Jelly Mesenchymal Stem Cells (hWJMSCs) on Apoptotic Gene Expression in a Breast Cancer Cell Line (MCF-7). Journal of Mathematics and Fundamental Sciences, 51(3); 205–224.
Widowati, W., D. Priyandoko, L. Lenny, R. Revika, S. Novianti, and H. S. Kusuma (2024). Camellia Sinensis L. Extract Suppresses Inflammation on Acute Respiratory Distress Syndrome Cell Models via Decreasing IL-1ß, IL-6, and COX-2 Expressions. Trends in Sciences, 21(1); 7010–7010.
Widowati, W., T. L. Wargasetia, F. Rahardja, R. F. Gunanegara, D. Priyandoko, and M. E. S. Gondokesumo (2023). hWJMSCs Inhibit Inflammation and Apoptosis in an ARDS Cell Model. Journal of Taibah University Medical Sciences, 18(6); 1519–1526.
Xie, J., Z. Lai, X. Zheng, H. Liao, Y. Xian, and Q. Li (2021). Apoptotic Activities of Brusatol in Human Non-Small Cell Lung Cancer Cells: Involvement of ROS-Mediated Mitochondrial-Dependent Pathway and Inhibition of Nrf2-Mediated Antioxidant Response. Toxicology, 451; 152680.
Yan, J., C. Ma, J. Cheng, Z. Li, and C. Liu (2015). HAX-1 Inhibits Apoptosis in Prostate Cancer Through the Suppression of Caspase-9 Activation. Oncology Reports, 34(5); 2776–2781.
Ye, G., M. Xu, Y. Shu, X. Sun, Y. Mai, and Y. Hong (2022). A Quassinoid Diterpenoid Eurycomanone From Eurycoma Longifolia Jack Exerts Anti-Cancer Effect Through Autophagy Inhibition. Molecules, 27(14); 4398.
Yunos, N. M., N. D. Amin, M. H. Jauri, S. K. Ling, N. H. Hassan, and N. J. Sallehudin (2022). The in Vitro Anti-Cancer Activities and Mechanisms of Action of 9-Methoxycanthin-6-One From Eurycoma Longifolia in Selected Cancer Cell Lines. Molecules, 27(3); 585.
Yunos, N. M., H. A. Wahab, M. G. Al-Thiabat, N. J. Sallehudin, and M. H. Jauri (2023). In Vitro and in Silico Analysis of the Anti-Cancer Effects of Eurycomanone and Eurycomalactone From Eurycoma Longifolia. Plants, 12; 2827.
Zeng, S., W. H. Shen, and L. Liu (2018). Senescence and Cancer. Cancer Translational Medicine, 4(3); 70–74.
Zhong, W., S. Dong, H. Wang, C. Pan, and S. Yang (2023). Functional Mechanism of MicroRNA-25-3p in Hilar Cholangiocarcinoma Cell Proliferation and Migration Through Regulation of Dual Specificity Phosphatase 5. Journal of Investigative Surgery, 36(1); 2202768.
Authors
Kania, N. ., Rahman, E. Y., Priyandoko, D., Sabrina, A. H. N. ., Widowati, W., Azis, R. ., Annaba, A., Hadiprasetyo, D. S. ., & Alexandro, G. . (2025). The Potential of Ethanol Extract of Pasak Bumi Roots (Eurycoma longifolia Jack) as an Anti-Prostate Cancer In Vitro Against PC-3 Cells. Science and Technology Indonesia, 10(2), 452–466. https://doi.org/10.26554/sti.2025.10.2.452-466
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