Cervical cancer is a significant health issue in Indonesia, with 36,633 cases reported in 2020. Chemotherapy, especially with doxorubicin, is a primary treatment but causes adverse effects such as cardiotoxicity, hair loss, nausea, vomiting, and immunosuppression. Combining chemotherapy with chemopreventive agents can enhance efficacy and reduce side effects. Selaginella doederleinii, known for its biflavonoid compounds, has potential chemopreventive activity. This study investigated the chemopreventive mechanism of Selaginella doederleinii ethanol extract (EESD) as a co-chemotherapy agent for HeLa cells through in silico and in vitro approaches. High-performance liquid chromatography analysis identified amentoflavone and 2,3-dihydro-3,3-diphenylapigenin as key compounds in EESD. Protein–protein interaction network analysis revealed 24 target receptors associated with cervical cancer, with ESR2 and HSP90AA1 being upregulated and involved in chemical carcinogenesis, estrogen signaling, cancer pathways, and endocrine resistance. EESD exhibited moderate cytotoxic activity (IC50 = 367.89 μg/ml) and demonstrated chemopreventive properties. The mechanism of EESD is presumed to involve the inhibition of key regulatory proteins in cervical cancer, namely ESR2 and HSP90AA1. Furthermore, EESD exhibited a slight synergistic effect with doxorubicin, with a combination index value of 0.85870. This study is the first to explore the chemopreventive potential of EESD along with its underlying mechanism, highlighting its role as a co-chemotherapy agent.
Ningrum DWC, Febriansah R, Juniananda M, Tasminatun S, Krisridwany A. Exploring the therapeutic potential of Selaginella doederleinii extract for cervical cancer treatment through network pharmacology and in vitro studies. J Appl Pharm Sci. 2025. Article in Press. http://doi.org/10.7324/JAPS.2026.237094
1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021;71(3):209–49. doi: https://doi.org/10.3322/caac.21660
2. Xing B, Guo J, Sheng Y, Wu G, Zhao Y. Human Papillomavirus-negative cervical cancer: a comprehensive review. Front Oncol. 2021;10:1–8. doi: https://doi.org/10.3389/fonc.2020.606335
3. Milliron BJ, Packel L, Dychtwald D, Klobodu C, Pontiggia L, Ogbogu O, et al. When eating becomes torturous: understanding nutrition-related cancer treatment side ffects among individuals with cancer and their caregivers. Nutrients. 2022;14(2). doi: https://doi.org/0.3390/nu14020356
4. Anggraini CY, Kusumaningtyas TA, Juniananda M, Ningrum DWC, Febriansah R, Hermawansyah A. In silico and in vitro study Selaginella doederleinii herb extract as an antineoplastic on MCF- 7 cells and formulation development of nano effervescent granule. Indones J Cancer Chemoprevention. 2023;14(2):128–38. doi: https://doi.org/10.14499/indonesianjcanchemoprev14iss2pp128-138
5. Ningrum DWC, Kusumaningtyas TA, Febriansah R, Juniananda M, Tasminatun S, Krisridwany A. Bioinformatics and molecular docking study of amentoflavone and 3,8-Biapigenin as inhibitors on cervical cancer proteins. Indones J Cancer Chemoprevention. 2023;14(2):105–16. doi: https://doi.org/10.14499/indonesianjcanchemoprev14iss2pp105-116
6. Muema FW, Liu Y, Zhang Y, Chen G, Guo M. Flavonoids from Selaginella doederleinii Hieron and their antioxidant and antiproliferative activities. Antioxidants. 2022;11(6):1–16. doi: https://doi.org/10.3390/antiox11061189
7. Noor F, Qamar MTU, Ashfaq UA, Albutti A, Alwashmi ASS, Aljasir MA. Network pharmacology approach for medicinal plants: review and assessment. Pharmaceuticals. 2022;15(5):1–33. doi: https://doi.org/10.3390/ph15050572
8. Lei X, Jing J, Zhang M, Guan B, Dong Z, Wang C. Bioinformatic identification of hub genes and analysis of prognostic values in colorectal cancer. Nutr Cancer. 2021;73(11–12):2568–78. doi: https://doi.org/10.1080/01635581.2020.1841249
9. Chuo SC, Nasir HM, Mohd-Setapar SH, Mohamed SF, Ahmad A, Wani WA, et al. A glimpse into the extraction methods of active compounds from plants. Crit Rev Anal Chem. 2022;52(4):667–96. doi: https://doi.org/10.1080/10408347.2020.1820851
10. Bennour N, Mighri H, Eljani H, Zammouri T, Akrout A. Effect of solvent evaporation method on phenolic compounds and the antioxidant activity of Moringa oleifera cultivated in Southern Tunisia. South African J Bot [Internet]. 2020;129:181–90. doi: https://doi.org/10.1016/j.sajb.2019.05.005
11. Li S, Yao H, Zhao M, Li Y, Huang L, Lin X. Determination of seven biflavones of Selaginella doederleinii by high performance liquid chromatography. Anal Lett. 2013;46(18):2835–45. doi: https://doi.org/10.1080/00032719.2013.831426
12. Wang X, Shen Y, Wang S, Li S, Zhang W, Liu X, et al. PharmMapper 2017 update : a web server for potential drug target identification with a comprehensive target pharmacophore database. Nucleic Acids Res. 2017;45:356–60. doi: https://doi.org/10.1093/nar/gkx374
13. Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-cepas J, et al. STRING v11 : protein – protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 2019;47:607–13. doi: https://doi.org/10.1093/nar/gky1131
14. Otasek D, Morris JH, Bouças J, Pico AR, Demchak B. Cytoscape automation: empowering workflow-based network analysis. Genome Biol. 2019; 1–1. doi: https://doi.org/10.1186/s13059-019-1758-4
15. Sherman BT, Hao M, Qiu J, Jiao X, Baseler MW, Lane HC, et al. DAVID: a web server for functional enrichment analysis and functional annotation of gene lists. Nucleic Acids Res. 2022;50:216– 21. doi: https://doi.org/10.1093/nar/gkac194
16. Chandrashekar DS, Karthikeyan SK, Korla PK, Patel H, Shovon AR, Athar M, et al. UALCAN: an update to the integrated cancer data analysis platform. Neoplasia (United States). 2022;25:18–27. doi: https://doi.org/10.1016/j.neo.2022.01.001
17. Jablonská E, Kubásek J, Vojt?ch D, Ruml T, Lipov J.Test conditions can significantly affect the results of in vitro cytotoxicity testing of degradable metallic biomaterials. Sci Rep. 2021;11(1):1–9. doi: https://doi.org/10.1038/s41598-021-85019-6
18. Ghasemi M, Turnbull T, Sebastian S, Kempson I. The MTT assay: utility, limitations, pitfalls, and interpretation in bulk and single-cell analysis. Int J Mol Sci. 2021;22(23):12827. doi: https://doi.org/10.3390/ijms222312827
19. Febriansah R, Komalasari T. Co-chemotherapeutic effect of Ageratum conyzoides L. chloroform fraction and 5-fluorouracil on hela cell line. Pharmacogn J.2019;11(5):913–8. doi: https://doi.org/10.5530/pj.2019.11.146
20. Huang DW, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 2009;4(1):44–57. doi: https://doi.org/10.1038/nprot.2008.211
21. Wang X, Huang X, Zhang Y. Involvement of human papillomaviruses in cervical cancer. Front Microbiol. 2018;9:2896. doi: https://doi.org/10.3389/fmicb.2018.02896
22. Romero-Masters JC, Lambert PF, Munger K. Molecular mechanisms of MmuPV1 E6 and E7 and implications for human disease. Viruses. 2022;14:2138. doi: https://doi.org/10.3390/v14102138
23. Chabner BA, Roberts TG Jr. Timeline: chemotherapy and the war on cancer. Nat Rev Cancer. 2005;5(1):65–72. doi: https://doi.org/10.1038/nrc1529
24. Conklin KA. Chemotherapy-associated oxidative stress: impact on chemotherapeutic effectiveness. Integr Cancer Ther. 2004;3:294– 300. doi: https://doi.org/10.1177/1534735404270335
25. Lee C, Longo VD. Fasting vs dietary restriction in cellular protection and cancer treatment: from model organisms to patients. Oncogene. 2011;30:3305–16. doi: https://doi.org/10.1038/onc.2011.91
26. Sui Y, Li S, Shi P, Wu Y, Li Y, Chen W, et al. Ethyl acetate extract from Selaginella doederleinii Hieron inhibits the growth of human lung cancer cells A549 via caspase-dependent apoptosis pathway. J Ethnopharmacol. 2016;190:261–71. doi: https://doi.org/10.1016/j.jep.2016.06.029
27. Tanaka T, Shimizu M, Kochi T, Moriwaki H. Chemical-induced carcinogenesis. J Exp Clin Med (Taiwan). 2013;5:203–9. doi: https://doi.org/10.1016/j.jecm.2013.10.009
28. Wang G, Yao S, Zhang XX, Song H. Rapid screening and structural characterization of antioxidants from the extract of Selaginella doederleinii Hieron with DPPH-UPLC-Q-TOF/MS method. Int J Anal Chem. 2015;2015. doi: https://doi.org/10.1155/2015/849769
29. Ullah Shirazi O, Muzaffar Ali Khan Khattak M, Azwani Mohd Shukri N, Mohd Nur Nasyriq MA, Shirazi O, Nur Nasyriq MA. Determination of total phenolic, flavonoid content and free radical scavenging activities of common herbs and spices. J Pharmacogn Phytochem JPP. 2014;104(33):104–8.
30. Yao H, Chen B, Zhang Y, Ou H, Li Y, Li S, et al. Analysis of the total biflavonoids extract from Selaginella doederleinii by HPLC-QTOF-MS and it’s in vitro and in vivo anticancer effects. Molecules. 2017;22(2):325. doi: https://doi.org/10.3390/molecules22020325
31. Mitra S, Lami MS, Ghosh A, Das R, Tallei TE, Fatimawali, et al. Hormonal therapy for gynecological cancers: how far has science progressed toward clinical applications?. Cancers. 2022;14:759. doi: https://doi.org/10.3390/cancers14030759
32. Lipowicz JM, Mali?ska A, Nowicki M, Raw?uszko-Wieczorek AA. Genes co-expressed with ESR2 influence clinical outcomes in cancer patients: TCGA data analysis. Int J Mol Sci. 2024;25(16):8707. doi: https://doi.org/10.3390/ijms25168707
33. DeFilippis RA, Goodwin EC, Wu L, DiMaio D. Endogenous Human Papillomavirus E6 and E7 proteins differentially regulate proliferation, senescence, and apoptosis in HeLa cervical carcinoma cells. J Virol. 2003;77(2):1551–63. doi: https://doi.org/10.1128/jvi.77.2.1551-1563.2003
34. Tusanti I, Johan A, Kisdjamiatun R. Sitotoksisitas in vitro ekstrak etanolik buah parijoto (Medinilla speciosa, reinw.ex bl.) terhadap sel kanker payudara T47D. J Gizi Indones (Indones J Nutr). 2014;2(2):53–8. doi: https://doi.org/10.14710/jgi.2.2.53-58
35. Bemmerlein L, Deniz IA, Karbanová J, Jacobi A, Drukewitz S, Link T, et al. Decoding single cell morphology in osteotropic breast cancer cells for dissecting their migratory, molecular and biophysical heterogeneity. Cancers (Basel). 2022;14(3):603. doi: https://doi.org/10.3390/cancers14030603
36. Sporn MB, Lippman SM. Agents for chemoprevention and their mechanism of action. Hamilton, ON: BC Decker;2003.
37. Allison KH, Hammond MEH, Dowsett M, McKernin SE, Carey LA, Fitzgibbons PL, et al. Estrogen and progesterone receptor testing in breast cancer: ASCO/CAP guideline update. J Clin Oncol. 2020;38(12). doi: https://doi.org/10.1200/JCO.19.02309
38. Grindstad T, Richardsen E, Andersen S, Skjefstad K, Rakaee khanehkenari M, Donnem T, et al. Progesterone receptors in prostate cancer: progesterone receptor B is the isoform associated with disease progression. Sci Rep. 2018;8(1):11358. doi: https://doi.org/10.1038/s41598-018-29520-5
39. Liu S, Fan W, Gao X, Huang K, Ding C, Ma G, et al. Estrogen receptor alpha regulates the Wnt/β-catenin signaling pathway in colon cancer by targeting the NOD-like receptors. Cell Signal. 2019;61:86–92. doi: https://doi.org/10.1016/j.cellsig.2019.05.009
40. Hiramitsu S, Ishikawa T, Lee WR, Khan T, Crumbley C, Khwaja N, et al. Estrogen receptor beta-mediated modulation of lung cancer cell proliferation by 27-hydroxycholesterol. Front Endocrinol (Lausanne). 2018;9:470. doi: https://doi.org/10.3389/fendo.2018.00470
41. Rades D, Setter C, Dahl O, Schild SE, Noack F. The prognostic impact of tumor cell expression of estrogen receptor-α, progesterone receptor, and androgen receptor in patients irradiated for nonsmall cell lung cancer. Cancer. 2012;118(1):157–63. doi: https://doi.org/10.1002/cncr.26282
42. Kawprasertsri S, Pietras RJ, Marquez-Garban DC, Boonyaratanakornkit V. Progesterone receptor (PR) polyproline domain (PPD) mediates inhibition of epidermal growth factor receptor (EGFR) signaling in non-small cell lung cancer cells. Cancer Lett. 2016;374(2):279–91. doi: https://doi.org/10.1016/j.canlet.2016.02.014
43. Song Q, Wen J, Li W, Xue J, Zhang Y, Liu H, et al. HSP90 promotes radioresistance of cervical cancer cells via reducing FBXO6- mediated CD147 polyubiquitination. Cancer Sci. 2022;113(4):1463– 74. doi: https://doi.org/10.1111/cas.15269
44. Sutejo IR, Putri H, Meiyanto E. Selektivitas Ekstrak Etanolik Buah Makassar (Brucea javanica) pada Kanker Payudara Metastasis secara in vitro. J Agromed Med Sci. 2016;2(1).
45. Tokarska-Schlattner M, Zaugg M, Zuppinger C, Wallimann T, Schlattner U. New insights into doxorubicin-induced cardiotoxicity: the critical role of cellular energetics. J Mol Cell Cardiol. 2006;41:9 . doi: https://doi.org/10.1016/j.yjmcc.2006.06.009
46. Baharuddin P, Satar N, Fakiruddin KS, Zakaria N, Lim MN, Yusoff NM, et al. Curcumin improves the efficacy of cisplatin by targeting cancer stem-like cells through p21 and cyclin D1-mediated tumour cell inhibition in non-small cell lung cancer cell lines. Oncol Rep. 2016;35(1):13–25. doi: https://doi.org/10.3892/or.2015.4371
47. Folkesson E, Niederdorfer B, Nakstad VT, Thommesen L, Klinkenberg G, Lægreid A, et al. High-throughput screening reveals higher synergistic effect of MEK inhibitor combinations in colon cancer spheroids. Sci Rep. 2020;10(1):11574. doi: https://doi.org/10.1038/s41598-020-77501-4
48. Calzetta L, Koziol-White C. Pharmacological interactions: synergism, or not synergism, that is the question. Curr Res Pharmacol Drug Discov. 2021;2:100046. doi: https://doi.org/10.1016/j.crphar.2021.100046
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