In vitro Anticancer Activities and GC-MS Analysis of the Leaf Ethanol Extracts from the Genus Piper (Piperaceae) in Lapuyan, Zamboanga del Sur, Philippines

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Published: 2025-07-01
DOI: https://doi.org/10.26538/tjnpr/v9i6.54
Keywords:
Ethnomedicinal, Ethnopharmacology, Natural product, Subanen

Main Article Content

Junnasir M. Sakilan
Jose Rizal Memorial State University1 – Dapitan City 7101, Philppines
Jhoan R.L. Pizon
Jose Rizal Memorial State University1 – Dapitan City 7101, Philppines
Cesar G. Demayo
Mindanao State University – Iligan Institute of Technology, Iligan City, Philippines
Praneet Opanasopit
Faculty of Pharmacy Silpakorn University, Nakhon Pathom 730000, Thailand

Abstract

The utilization of some of the Piper species in traditional medicine in cancer treatment suggests that the genus is a promising source of new anticancer compounds. The current study aims to determine the cytotoxic activity of the collected Piper species leaf extracts against three cancer cell lines using the MTT assay and to identify the major compounds accounting for the activity using GC-MS. The leaf ethanol extracts of the studied Piper species exhibited potent cytotoxic activity against three human cancer cell lines, represented by cervical carcinoma (HeLa), breast Cancer (MCF-7), and colon adenocarcinoma (HT-29). It was observed that P. betle has the lowest
IC50 value for HeLa and HT-29, while P. caninum for MCF-7 with a value of 20.24 ± 1.26 μg/mL, 23.51 ± 4.50 μg/mL, and 27.34 ± 3.13 μg/mL, respectively. This suggests that P. betle is an effective agent for both HeLa and HT-29 cancer cell lines, while P. caninum has higher potency for MCF-7. Analysis of the bioactive compounds of the Piper ethanol leaf extracts using GC-MS analysis revealed the presence of anticancer compounds. It is suggested that further studies, like the isolation of individual compounds, the analysis of their bioactivity, and the understanding of their toxicity profile, will produce significant results. Finally, the study results show that the ethnomedicinal application of Piper species by the Subanen of Lapuyan, Zamboanga del Sur, has a pharmacological basis.

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Vol. 9 No. 6 (2025): Tropical Journal of Natural Product Research (TJNPR)

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Articles
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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

How to Cite

Sakilan, J. M., Pizon, J. R., Demayo, C. G., & Opanasopit, P. (2025). In vitro Anticancer Activities and GC-MS Analysis of the Leaf Ethanol Extracts from the Genus Piper (Piperaceae) in Lapuyan, Zamboanga del Sur, Philippines. Tropical Journal of Natural Product Research (TJNPR), 9(6), 2755-2763. https://doi.org/10.26538/tjnpr/v9i6.54

References

1. World Health Organization (WHO). World Cancer Report 2014. International agency for research on cancer, Lyon.

2. Süntar I. Importance of ethnopharmacological studies in drug discovery: role of medicinal plants. Phytochemistry Reviews, 2020; 19(5): 1199–1209. [https://doi.org/10.1007/s11101-019-09629-9](https://doi.org/10.1007/s11101-019-09629-9)

3. Cragg GM, Grothaus PG, and Newman DJ. New Horizons for Old Drugs and Drug Leads. J. Nat. Prod. 2014; 77(3): 703-723.

4. Raut JS, Karuppayil SM. A status review on the medicinal properties of essential oils. Ind. Crops Prod. 2014; 62: 250–264.

5. Dyer LA, Palmer AND. (Eds.). Piper: A model genus for studies of phytochemistry, ecology, and evolution. Kluwer Academic/Plenum Publishers, New York. 2004.

6. Picard G, Valadeau C, Albán–Castillo J, Rojas R, Starr JR, Callejas-Posada R, Bennett SAL, Arnason JT. Assessment of in vitro pharmacological effect of Neotropical Piperaceae in GABAergic bioassays in relation to plants traditionally used for folk illness by the Yanesha (Peru). J. Ethnopharm. 2014; 155: 1500–1507.

7. Hardoko H, Jessica M, Halim Y. Antibacterial Activity of Cough Suppressant Functional Drink from Betel Leaves (Piper sarmentosum), Lime (Citrus aurantifolia), and Honey. Trop J Nat Prod Res. 2020; 4(10): 695–702. [https://doi.org/10.26538/tjnpr/v4i10.7](https://doi.org/10.26538/tjnpr/v4i10.7)

8. Wangpan T, Chetry LB, Tsering J, Taka T, Tangjang S. Anti-Malarial Plants of Jonai, India: An Ethnobotanical Approach. Not Sci Biol. 2016; 8(1): 27–32. [https://doi.org/10.15835/nsb.8.1.9750](https://doi.org/10.15835/nsb.8.1.9750)

9. Daorueang D. The Protective Effect of Sa-khan (Piper, Piperaceae) Against High Glucose-Induced Cytotoxicity in Human Proximal Tubular Epithelial Cells. Trop J Nat Prod Res. 2022; 6(12): 1953–1956. [https://doi.org/10.26538/tjnpr/v6i12.9](https://doi.org/10.26538/tjnpr/v6i12.9)

10. Hashim NA, Ahmad F, Jani NA, Susanti D. In vitro Antioxidant, Antityrosinase, Antibacterial and Cytotoxicity Activities of the Leaf and Stem Essential Oil from Piper magnibaccum C. DC. J Essent Oil-Bear Plants. 2017; 20(1): 223–232. [https://doi.org/10.1080/0972060X.2017.1282839](https://doi.org/10.1080/0972060X.2017.1282839)

11. Abdullah NA, Zain M, Hamid NA, Ramli NW. Essential oil from Piperaceae as a potential for biopesticide agents: A review. Food Research. 2020; 4(S5): 1–10. [https://doi.org/10.26656/fr.2017.4(S5).007](https://doi.org/10.26656/fr.2017.4%28S5%29.007)

12. Ahmad I, Ambarwati NSS, Arifuddin M, Rijai L, Mun’im A. Pharmacognostic and Cytotoxicity Evaluation of Indonesia Native Plant of Piper acre Blume Leaves (Piperaceae). Pharmacogn J. 2017; 9(3): 400–404.

13. Quijano MA, Calleja RP, Miranda DE. Areas of Endemism and Distribution for Neotropical Piper species (Piperaceae). J Biogeogr. 2006; 33: 1266–1278. [https://doi.org/10.1111/j.1365-2699.2006.01501.x](https://doi.org/10.1111/j.1365-2699.2006.01501.x)

14. De Candolle C. A revision of Philippine Piperaceae. Philipp J Sci, Bot. 1910; 5: 405–463.

15. Suwanphakdee C, Simpson DA, Hodkinson TR, Chantaranothai P. Taxonomic notes on the genus Piper (Piperaceae). Nordic J Bot. 2016; 34(5): 605–618. [https://doi.org/10.5061/dryad.qp50f](https://doi.org/10.5061/dryad.qp50f)

16. Aureo WA, Reyes Jr TD. Composition and community structure of plant species in a secondary growth forest in the central Philippines. J Biodivers Conserv Bioresour Manag. 2023; 9(1): 13–28.

17. Gardner RO. Piper in the Philippine Islands: the climbing species. Blumea. 2006; 51(3).

18. Dongre U, Meshram T, Dighe S, Narnawre K, Mehere B, Somkuwar SR. Screening of Selected Ethno-Medicinal Plants for Anticancer Activity. Advances in Zoology and Botany. 2020; 8(5): 447–452. [https://doi.org/10.13189/azb.2020.080509](https://doi.org/10.13189/azb.2020.080509)

19. Wang YH, Morris-Natschke SL, Yang J, Niu HM, Long CL, Lee KH. Anticancer Principles from Medicinal Piper (胡椒 Hú Jiāo) Plants. J Tradit Complement Med. 2014; 4(1): 8–16.

20. Sidiq LO, Segun PA, Ogbole OO. Total phenolic contents and antioxidant activity of nine medicinal plants used in Nigerian traditional medicine. Trop J Nat Prod Res. 2018; 2(9): 438–441. [https://doi.org/10.26538/tjnpr/v2i9.6](https://doi.org/10.26538/tjnpr/v2i9.6)

21. Matowa PR, Gundidza M, Gwanzura L, Nhachi CFB. A survey of ethnomedicinal plants used to treat cancer by traditional medicine practitioners in Zimbabwe. BMC Complementary Medicine and Therapies. 2020; 20(1): 1–13. [https://doi.org/10.1186/s12906-020-03046-8](https://doi.org/10.1186/s12906-020-03046-8)

22. Dapar MLG, Meve U, Liede-Schumann S, Alejandro GJD. Ethnomedicinal plants used for the treatment of cuts and wounds by the agusan manobo of sibagat, agusan del sur, philippines. Ethnobot. Res. Appl. 2020; 19. [https://doi.org/10.32859/era.19.31.1-18](https://doi.org/10.32859/era.19.31.1-18)

23. Greenshields AL, Doucette CD, Sutton KM, Madera L, Annan H, Yaffe PB, Knickle AF, Dong Z, Hoskin DW. Piperine inhibits the growth and motility of triple-negative breast cancer cells. Cancer Letters. 2015; 357(1): 129–140.

24. Meegan MJ, Nathwani S, Twamley B, Zisterer DM, O’Boyle NM. Piper longumine (piplartine) and analogues: Antiproliferative microtubule-destabilising agents. Eur. J. Med. Chem. 2017; 125: 453–463.

25. Sakilan JM, Demayo CG, Opanasopit P. Phytochemical Analysis and Determination of Antimicrobial, Antioxidant and Anticancer Activity of the Leaf Ethanolic Extracts of Piper Sarmentosum Roxb. in Lapuyan Zamboanga Del Sur, Philippines. Int. J. Pharm. Sci. Res. 2019; 10(12): 5715–5722. [https://doi.org/10.13040/IJPSR.0975-8232.10(12).5715-22](https://doi.org/10.13040/IJPSR.0975-8232.10%2812%29.5715-22)

26. Hematpoor A, Paydar M, Liew SY, Sivasothy Y, Mohebali N, Looi CY, Wong WF, Azirun MS, Awang K. Phenylpropanoids isolated from Piper sarmentosum Roxb. induce apoptosis in breast cancer cells through reactive oxygen species and mitochondrial-dependent pathways. Chemico-Biological Interactions. 2018; 279: 210–218.

27. Rossette MC, Moraes DC, Sacramento EK, Romano-Silva MA, Carvalho JL, Gomes DA, Caldas H, Friedman E, Bastos-Rodrigues L, De Marco L. The In Vitro and In Vivo Antiangiogenic Effects of Flavokawain B. Phytotherapy Research. 2017; 31(10): 1607–1613.

28. Silva MA, Passarini GM, Martinez LN, Facundo VA, Teles CGB, Kuehn CC. Chemical Constituents and Bioactivities of Essential Oils From. 2020; 776–817.

29. Carvalho ÊS, Ayres VFS, Oliveira MR, Corrêa GM, Takeara R, Guimarães AC, Santiago MB, Oliveira TAS, Martins CHG, Crotti AEM, Silva EO. Anticarcinogenic Activity of Three Essential Oils from Brazilian Piperaceae. Pharmaceuticals. 2022; 15(8): 1–11. [https://doi.org/10.3390/ph15080972](https://doi.org/10.3390/ph15080972)

30. Pizon JRL, Nuñeza OM, Uy MM, Senarath WTPSK. Ethnobotany of Medicinal Plants Used by the Subanen Tribe of Lapuyan, Zamboanga del Sur. Bull. Env. Pharmacol. Life Sci. 2016; 5(5): 53–67.

31. Fajardo WT, Cancino LT, Dudang EB, De Vera LA, Pambid RM, Junio AD. Ethnobotanical Study of Traditional Medicinal Plants Used by Indigenous Sambal-Bolinao of Pangasinan, Philippines. PSU J. Nat. Sci. Allied Sci. 2017; 1(1): 52–63.

32. Angagan JS, Buot Jr IE, Relox RE, Rebancos CM. Ethnobotany of The Plant Resources in Conner, Apayao, Northern Luzon, Philippines. J. Nat. Stud. 2010; 9(1): 31–38.

33. Ducusin MB. Ethnomedicinal Knowledge of Plants among the Indigenous Peoples of Santol, La Union, Philippines. Electronic Journal of Biology. 2017; 13(4): 360–382.

34. Olowa L, Demayo CG. Ethnobotanical Uses of Medicinal Plants among the Muslim Maranaos in Iligan City, Mindanao, Philippines. Adv. Environ. Biol. 2015; 9(27): 204–215.

35. Morilla LJG, Sumaya NHN, Rivero HI, Madamba MRSB. Medicinal Plants of the Subanens in Dumingag, Zamboanga del Sur, Philippines. International Conference on Food, Biological and Medical Sciences (FBMS-2014) Jan. 28–29, 2014 Bangkok (Thailand).

36. Freshney RI. Culture of Animal Cells, A Manual of Basic Techniques, Wiley-Liss, Inc., New York, USA. 2000.

37. Chipiti T, Ibrahim MA, Koorbanally NA. In-vitro antioxidant activity and GC-MS analysis of the ethanol and aqueous extracts of Cissus cornifolia (Baker) Splanch (Vitaceae) parts. Acta Pol Pharm. 2015; 72: 119–127.

38. Rungrojsakul M, Okonogi S, Panyajai P, Anuchapreeda S. Cytotoxic and antiproliferative effects of crude ethanolic extract from Piper betle leaves on leukemic cell lines. J Assoc Med Sci. 2021; 54(2): 42–50.

39. Gundala SR, Yang C, Mukkavilli R, Paranjpe R, Brahmbhatt M, Pannu V, Cheng A, Reid MD, Aneja R. Hydroxychavicol, a betel leaf component, inhibits prostate cancer through ROS-driven DNA damage and apoptosis. Toxicol. Appl. Pharmacol. 2014; 280(1): 86–96. [https://doi.org/10.1016/j.taap.2014.07.012](https://doi.org/10.1016/j.taap.2014.07.012)

40. Veettil SR, Sunil EA, Mukunda A, Mohan A, John S, Pynadath MK. Anticancer effect of Piper betle leaf extract on KB cell lines – an in vitro study. Oral Maxillofac Pathol J. 2022; 13(1): 28–31.

41. Paarakh PM, Sreeram DC, Shruthi SD, Ganapathy SPS. In vitro cytotoxic and in silico activity of Piperine isolated from Piper nigrum fruits Linn. In Silico Pharmacology. 2015; 3–9. [https://doi.org/10.1186/s40203-015-0013-2](https://doi.org/10.1186/s40203-015-0013-2)

42. Mayanga-Herrera A, Tapia-Rojas S, Fukusaki-Yoshizawa A, Marcelo-Rodríguez Á, Amiel-Pérez J. Cytotoxic activity of the chloroform fraction of Piper aduncum and its effect on the cell cycle in gastric cancer cell lines. Revista Peruana de Medicina Experimental y Salud Pública. 2020; 37(3): 471–477. [https://doi.org/10.17843/rpmesp.2020.373.5157](https://doi.org/10.17843/rpmesp.2020.373.5157)

43. U.S. Department of Agriculture, Agricultural Research Service. Dr. Duke's Phytochemical and Ethnobotanical Databases. 1992–2016. [https://phytochem.nal.usda.gov](https://phytochem.nal.usda.gov)

44. Mgbeahuruike EE, Yrjonen T, Vuorela H, Holm Y. Bioactive compounds from medicinal plants: Focus on Piper species. South African Journal of Botany. 2017; 112: 54–69.

45. Narayanamoorthi V, Vasantha K, Rency RC, Maruthasalam A. GC MS determination of bioactive components of Peperomia pellucida (L.) Kunth. Bioscience Discovery. 2016; 6(2): 83–88.

46. Ghazali NF, Mohd MA, Ibrahim MA, Muhammad TST. Phytochemical and pharmacological profile of Kaduk (Piper sarmentosum Roxb.). Malaysian Nature Journal. 2018; 70(2): 195–202.

47. Zhou W, Bai M, Huang X, Song S. Amide Alkaloids from the genus Piper: A Review of its Phytochemistry. Asian Journal of Traditional Medicine. 2018; 13(3).

48. Saxena M, Saxena J, Nema R, Singh D, Gupta A. Phytochemistry of Medicinal Plants. Journal of Pharmacognosy and Phytochemistry. 2013; 1(6).

49. Jesonbabu J, Spandana N, Aruna Lakshmi K. The Potential Activity of Hydroxychavicol against Pathogenic Bacteria. Journal of Bacteriology and Parasitology. 2011; 2:121. [https://doi.org/10.4172/2155-9597.1000121](https://doi.org/10.4172/2155-9597.1000121)

50. Costa SB, Kreutz T, Limberger RP, Teixeira HF, Veiga-Junior VF, Koester LS. Piper aduncum Essential Oil Rich in Dillapiole: Development of Hydrogel-Thickened Nanoemulsion and Nanostructured Lipid Carrier Intended for Skin Delivery. Pharmaceutics. 2022; 14(11): 2525.

51. Luyen BTT, Thao NP, Widowati W, Fauziah N, Maesaroh M, Herlina T, Kim YH. Chemical constituents of Piper aduncum and their inhibitory effects on soluble epoxide hydrolase and tyrosinase. Medicinal Chemistry Research. 2016. [https://doi.org/10.1007/s00044-016-1735-3](https://doi.org/10.1007/s00044-016-1735-3)

52. Monzote L, Scull R, Cos P, Setzer WN. Essential Oil from Piper aduncum: Chemical Analysis, Antimicrobial Assessment, and Literature Review. Medicines. 2017; 4(3): 49. [https://doi.org/10.3390/medicines4030049](https://doi.org/10.3390/medicines4030049)

53. Ulia RV, Suryati, Santoni A. Cytotoxic Potential of Essential Oil Isolated from Clibadium surinamense L. Leaves Against T47D Breast and HeLa Cervical Cancer Cells. Molekul. 2023; 18(2): 289–299. [https://doi.org/10.20884/1.jm.2023.18.2.7816](https://doi.org/10.20884/1.jm.2023.18.2.7816)

54. Pavithra PS, Mehta A, Verma RS. Aromadendrene oxide 2 induces apoptosis in skin epidermoid cancer cells through ROS-mediated mitochondrial pathway. Life Sciences. 2018; 197: 19–29. [https://doi.org/10.1016/j.lfs.2018.01.029](https://doi.org/10.1016/j.lfs.2018.01.029)

55. Ferreira AK, de-Sa-Junior PL, Pasqualoto KF, de Azevedo RA, Camara DA, Costa AS, Matsuo AL, Massaoka MH, Auada AV, Lebrun I, Damilao MC, Tavares MT, Magri FM, Kerkis I, Filho RP. Cytotoxic effects of dillapiole on MDA-MB-231 cells involve the induction of apoptosis through the mitochondrial pathway by inducing oxidative stress while altering the cytoskeleton network. Biochimie. 2014; 99: 195–207. [https://doi.org/10.1016/j.biochi.2013.12.008](https://doi.org/10.1016/j.biochi.2013.12.008)

56. Hago S, Lu T, Abdelgadir AA, Yassin S, Ahmed EM, Xu H. Phytochemical Constituents and In-Vitro Anticancer Activity of some Medicinal Plant Extracts against MCF-7 and MDA-MB-435 Human Breast Cancer Cell Lines. Trop J Nat Prod Res. 2023; 7(3): 2506–2515. [https://doi.org/10.26538/tjnpr/v7i3.6](https://doi.org/10.26538/tjnpr/v7i3.6)

57. Assunção JAS, Machado DB, Felisberto JS, Chaves DSA, Campos DR, Cid YP, et al. Insecticidal activity of essential oils from Piper aduncum against Ctenocephalides felis felis: a promising approach for flea control. Rev Bras Parasitol Vet. 2024; 33(3): e007624. [https://doi.org/10.1590/S1984-29612024050](https://doi.org/10.1590/S1984-29612024050)

58. Volpe HX, Fazolin M, Garcia RB, Magnani RF, Barbosa JC, Miranda MP. Efficacy of essential oil of Piper aduncum against nymphs and adults of Diaphorina citri. Pest Manag Sci. 2016; 72: 1242–1249.

59. Hui LM, Zhao GD, Zhao JJ. δ-Cadinene inhibits the growth of ovarian cancer cells via caspase-dependent apoptosis and cell cycle arrest. Int J Clin Exp Pathol. 2015; 8(6): 6046–6056.

60. Singh D, Gupta R, Saraf SA. Herbs-are-the-dynamic-agents-in-cancer-therapy: a-review. Journal of Herbal Medicine and Toxicology. 2011; 5(2): 19–30.