Evaluation of Garcinia cowa Leaf Extract as A Potential Anticancer Agent: Cytotoxicity, Selectivity, and Apoptotic Effects on MCF-7/HER-2 Cells
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Abstract
The global burden of cancer highlights the urgent need for therapeutic agents that are both effective and selective, with natural plant-derived compounds offering promising medicinal potential.This study aims to evaluate the anticancer potential of Garcinia cowa Roxb. leaf extract, a plant known for its medicinal properties, especially the cytotoxic effect, cell cycle inhibition, apoptosis in MCF-7/HER-2 breast cancer cell lines, and selectivity against normal Vero cells. The cytotoxic activity of G. cowa leaf extract against MCF-7/HER-2 cell lines was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry was conducted at the extract’s IC50 concentration for cell cycle modulation and apoptosis analysis, and the results were analysed using a FACSCalibur flow cytometer. G. cowa leaf extract demonstrated strong cytotoxic activity with an IC50 of 15.68 μg/mL against MCF-7/HER-2 cell lines and 303.6 μg/mL against normal Vero cells, yielding a selectivity index above 3. The extract significantly altered the MCF-7/HER-2 cell cycle, reducing G0-G1 phase cells from 38.1% to 29.9% and increasing the S phase (7.5% to 9.7%) and G2-M phase (49.5% to 55.3%), indicating cell cycle arrest. Additionally, the extract reduced early apoptosis from 5.5% to 2.7% and increased late apoptosis from 5.4% to 12.2%, while necrosis rose from 3.8% to 14.6%, reflecting enhanced apoptosis, necrosis, and cell accumulation in the S and G2-M phases. G. cowa leaf extract showed strong anticancer potential against MCF-7/HER-2 cells by disrupting the cell cycle and promoting apoptosis and necrosis, with high selectivity for cancer cells.
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Observatory GC. Cancer Today [Online]. International Agency for Research on Cancer. 2023 [cited 2023 Sep 9]. Available from: https://gco.iarc.fr
Freudenberg JA, Wang Q, Katsumata M, Drebin J, Nagatomo I, Greene MI. The role of HER2 in early breast cancer metastasis and the origins of resistance to HER2-targeted therapies. Exp. Mol. Pathol. 2009; 87(1):1-11.
Kong X, Zhang K, Wang X, Yang X, Li Y, Zhai J, Xing Z, Qi Y, Gao R, Feng X, Wang J. Mechanism of trastuzumab resistance caused by HER-2 mutation in breast carcinomas. Cancer Manag Res. 2019; 5971-5982.
Susanti M, Pratama AR, Suryani MI. Development and validation of TLC-densitometry method for quantification of tetraprenyltoluquinone in the stem bark hexane extract of Garcinia cowa Roxb. Heliyon. 2022; 8(9):1-6.
Ritthiwigrom T, Laphookhieo S, Pyne SG. Chemical constituents and biological activities of Garcinia cowa Roxb. Maejo Int. J. Sci. Technol. 2013, 7(02), 212-231.
Husni E, Nahari F, Wirasti Y, Wahyuni FS. Cytotoxicity study of ethanol extract of the stem bark of asam kandis (Garcinia cowa Roxb.) on T47D breast cancer cell line. Asian Pac. J. Trop. Biomed. 2015; 5(3):249-252.
Wahyuni FS, Febria S, Arisanty D. Apoptosis induction of cervical carcinoma HeLa cells line by dichloromethane fraction of the rinds of Garcinia cowa Roxb. Phcog J. 2017; 9(4):475-477.
Wahyuni FS, Shaari K, Stanslas J, Lajis NH, Dachriyanus H. Cytotoxic xanthones from the stem bark of Garcinia cowa Roxb. J Chem Pharm Res. 2015; 7(1):227-236.
Wahyuni FS, Syafri S, Permatasari D, Hefni D, Hamidi D, Nomiza NA, Rezti IA. Cowanin induces apoptosis in breast cancer cells via Bcl-2 signalling pathway. J Altern Complement Med. 2023; 20(3):631-636.
Chouni A, Pal A, Gopal PK, Paul S. GC-MS analysis and screening of antiproliferative potential of methanolic extract of Garcinia cowa on different cancer cell lines. Phcog J. 2021; 13(2):347-361.
Wahyuni FS, Triastuti DH, Arifin H. Cytotoxicity study of ethanol extract of the leaves of asam kandis (Garcinia cowa Roxb.) on T47D breast cancer cell line. Phcog J. 2015; 7(6):369-371.
Furqan M, Dachriyanus, Susanti M, Putra PP, Wahyuni FS. Unravelling the interaction between Garcinisidone-a and HER-2 protein in breast cancer: a computational study. Int J App Pharm. 2024; 16(Special Issue 1):99–104.
Wahyuni FS, Putri DE, Putra YU, Hamidi D. Cytotoxic activity of Taxus sumatrana (miq.) De laub. Bark, leaves, and shoots on hela, T47D, and MCF-7/HER2 cell lines Int J App Pharm. 2024; 16(Special Issue 1):93–98.
Coecke S, Balls M, Bowe G, Davis J, Gstraunthaler G, Hartung T, Hartung T, Hay R, Merten OW, Price A, Schechtman L, Stacey G. Guidance on good cell culture practice: A Report of the Second ECVAM Task Force on good cell culture practice. Atla-Altern Lab Anim. 2005; 33(3):261–287.
Hefni D, Wahyuni FS. The Cytotoxicity Study of Lantana camara Linn Essential Oil on HeLa Cancer Cells Line. Phcog J. 2021; 13(6):1498-1501.
Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods. 1983; 65(1-2):55-63.
Dewi IP, Dachriyanus YA, Ismail NH, Hefni D, Susanti M, Syafri S, Wahyuni FS. Curcuma Aeruginosa Roxb. Extract Inhibits the Production of Proinflammatory Cytokines on Raw 264.7 Macrophages. Int J App Pharm. 2024; 16(1):41-44.
Rahmawati N, Ismail NH, Hamidi D, Wahyuni FS. Cytotoxic Activity Screening of Various Uncaria Spp Plants on T47D Breast Cancer Cells. Trop. J. Nat. Prod. Res. 2023; 7(1):2218-2221
Ibrahim OHM, Abo-Elyousr KAM, Asiry KA, Alhakamy NA, Mousa MAA. Phytochemical Characterisation, Antimicrobial Activity and In Vitro Antiproliferative Potential of Alchemilla vulgaris Auct Root Extract against Prostate (PC-3), Breast (MCF-7) and Colorectal Adenocarcinoma (Caco-2) Cancer Cell Lines. Plants. 2022; 11(16):1–14.
Qanash H, Bazaid AS, Aldarhami A, Alharbi B, Almashjary MN, Hazzazi MS, Felemban HR, Abdelghany TM. Phytochemical characterisation and efficacy of Artemisia judaica extract loaded chitosan nanoparticles as inhibitors of cancer proliferation and microbial growth. Polymers. 2023; 15(2):1-19.
Weerapreeyakul N, Junhom C, Barusrux S, Thitimetharoch T. Induction of apoptosis in human hepatocellular carcinoma cells by extracts of Lannea coromandelica (Houtt.) Merr. and Diospyros castanea (Craib) Fletcher. Chin Med. 2016; 11:1-10.
Febriansah R, Putri DDP, Sarmoko, Nurulita NA, Meiyanto E, Nugroho AE. Hesperidin as a preventive resistance agent in MCF-7 breast cancer cells line resistance to doxorubicin. Asian Pac J Trop Biomed. 2014; 4(3):228–233.
Lestari B, Novitasari D, Putri H, Haryanti S, Sasmito E, Meiyanto E. Evaluation of The Genotoxicity of Three Food Additives using CHO-K1 Cells under in vitro Micronucleus Flow Cytometry Assay. Indones. J. Cancer Chemoprevention. 2017; 8(2):74-80.
Ionescu CA, Aschie M, Matei E, Cozaru GC, Deacu M, Mitroi AF, Baltatescu GI, Nicolau AA, Mazilu L, Tuta LA, Iorga IC. Characterisation of the Tumor Microenvironment and the Biological Processes with a Role in Prostatic Tumorigenesis. Biomedicines. 2022; 10(7):1-20.
Satria D, Silalahi J, Haro G, Ilyas S, Hasibuan PAZ. Cell cycle inhibition of ethylacetate fraction of Zanthoxylum acanthopodium DC. Fruit against T47D cells. Open Access Maced J Med Sci. 2019; 7(5):726–729.
Engel N, Oppermann C, Falodun A, Kragl U. Proliferative effects of five traditional Nigerian medicinal plant extracts on human breast and bone cancer cell lines. J Ethnopharmacol. 2011; 137(2):1003–1010.
Chatri M, Hasibuan PA, Meiyanto E, Putra DP, Septisetyani EP, Satria D, Waruwu SB. Effect of African Leaves (Vernonia amygdalina Delile) on the Development of T47D Breast Cancer Cells. Trop J Nat Prod Res. 2024; 8:7740–7746.
Engel N, Falodun A, Kühn J, Kragl U, Langer P, Nebe B. Pro-apoptotic and anti-adhesive effects of four African plant extracts on the breast cancer cell line MCF-7. BMC Complement Altern Med. 2014; 14(1):1–13.
Putri RH, Soetrisno, Wasita B, Priyanto H, Pamungkasari EP, Cilmiaty R. Investigating the Effects of the Ethyl Acetate Fraction of Mas Banana Bracts: Suppressing Cell Growth, Promoting Cell Death, and Targeting Specific Molecules in Hela Cells. Trop J Nat Prod Res. 2024; 8(11):9035–9042.
Ambrosio MR, Magli E, Caliendo G, Sparaco R, Massarelli P, D’Esposito V, Migliaccio T, Mosca G, Fiorino F, Formisano P. Serotoninergic receptor ligands improve Tamoxifen effectiveness on breast cancer cells. BMC Cancer. 2022; 22(1):1–14.
Eren CY, Gurer HG, Gursoy OO, Sezer CV. Antitumor Effects of L-citrulline on Hela Cervical Cancer Cell Lines. Anticancer Agents Med Chem. 2022; 22(18):3157–3162.
Hefni D, Dachriyanus, Nahda AM, Wahyuni FS. Effect of Cowanin on Cyclin D1 Expression In MCF-7/HER2 Breast Cancer Cells. Int J App Pharm. 2024; 16(Special Issue 1):149–151.
Sangour MH, Ali IM, Atwan ZW, Al Ali AAALA. Effect of Ag nanoparticles on viability of MCF-7 and Vero cell lines and gene expression of apoptotic genes. Egypt. J. Med. Hum. Genet. 2021; 22(1):1–11.
Widiandani T, Tandian T, Zufar BD, Suryadi A, Purwanto BT, Hardjono S. In vitro study of pinostrobin propionate and pinostrobin butyrate: Cytotoxic activity against breast cancer cell T47D and its selectivity index. J Public Health Afr. 2023; 14:1-6.
Calderón-Montaño JM, Martínez-Sánchez SM, Jiménez-González V, Burgos-Morón E, Guillén-Mancina E, Jiménez-Alonso JJ, Díaz-Ortega P, García F, Aparicio A, López-Lázaro M. Screening for selective anticancer activity of 65 extracts of plants collected in Western Andalusia, Spain. Plants. 2021; 10(10):2193-2122.
Jabit ML, Wahyuni FS, Khalid R, Israf DA, Shaari K, Lajis NH, Stanslas J. Cytotoxic and nitric oxide inhibitory activities of methanol extracts of Garcinia species. Pharm Biol. 2009; 47(11):1019–1026.
Cornwell JA, Crncec A, Afifi MM, Tang K, Amin R, Cappell SD. Loss of CDK4/6 activity in S/G2 phase leads to cell cycle reversal. Nature. 2023; 619(7969):363–370.
Kumar P, Koach J, Nekritz E, Mukherjee S, Braun BS, DuBois SG, Nasholm N, Haas-Kogan D, Matthay KK, Weiss WA, Gustafson C. Aurora Kinase A inhibition enhances DNA damage and tumor cell death with 131I-MIBG therapy in high-risk neuroblastoma. EJNMMI research. 2024; 14(1):54-66.
Zdioruk M, Want A, Mietelska-Porowska A, Laskowska-Kaszub K, Wojsiat J, Klejman A, Użarowska E, Koza P, Olejniczak S, Pikul S, Konopka W. A new inhibitor of tubulin polymerisation kills multiple cancer cell types and reveals p21-mediated mechanism determining cell death after mitotic catastrophe. Cancers. 2020; 12(8):1-21.
Jenie RI, Handayani S, Susidarti RA, Udin LZ, Meiyanto E. The cytotoxic and antimigratory activity of Brazilin-doxorubicin on MCF-7/HER2 cells. Adv Pharm Bull. 2018; 8(3):507–516.
Kusumastuti R, Utomo RY, Khumaira A, Putri H, Jenie RI, Meiyanto E. Pentagamaboronon-0 increased cytotoxicity of and inhibited metastasis induction by doxorubicin in breast cancer cells. J Appl Pharm Sci. 2019; 9(6):43–51.
Mohamed SIA, Elsayed GH, El Shaffai A, Yahya SMM, Mettwally WSA. In-vitro study of cytotoxic and apoptotic potential of Thalassia hemprichii (Ehren.) Asch. and Enhalus acoroides (L.f.) Royle against human breast cancer cell line (MCF-7) with correlation to their chemical profile. BMC Complement Med Ther. 2024; 24(1):1–16.
Chottanapund S, Van Duursen MB, Zwartsen A, Timtavorn S, Navasumrit P, Kittakoop P, Sureram S, Ruchirawat M, Van den Berg M. Depsidones inhibit aromatase activity and tumor cell proliferation in a co-culture of human primary breast adipose fibroblasts and T47D breast tumor cells. Toxicol Rep. 2017; 4:165-171.
Zwartsen A, Chottanapund S, Kittakoop P, Navasumrit P, Ruchirawat M, Van Duursen MB, Van den Berg M. Evaluation of anti-tumour properties of two depsidones–Unguinol and Aspergillusidone D–in triple-negative MDA-MB-231 breast tumour cells. Toxicol Rep. 2019; 6:1216-1222.
Sun J, Li M, Lin T, Wang D, Chen J, Zhang Y, Mu Q, Su H, Wu N, Liu A, Yu Y. Cell cycle arrest is an important mechanism of action of compound Kushen injection in the prevention of colorectal cancer. Sci Rep. 2022; 12(1):1-16.
Vogel C, Hager C, Bastians H. Mechanisms of mitotic cell death induced by chemotherapy-mediated G 2 checkpoint abrogation. Cancer Res. 2007; 67(1):339–345.
Wiecek AJ, Cutty SJ, Kornai D, Parreno-Centeno M, Gourmet LE, Tagliazucchi GM, Jacobson DH, Zhang P, Xiong L, Bond GL, Barr AR. Genomic hallmarks and therapeutic implications of G0 cell cycle arrest in cancer. Genome Biol. 2023; 24(1):1–35.