Cisplatin-Vernonia amygdalina combination on pancreatic cancer

Excerpt:


J. Pharm. Pharmacogn. Res., vol. 12, no. 5, pp. 870-880, Sep-Oct 2024. DOI: https://doi.org/10.56499/jppres23.1748_12.5.870 Original Article Combination of cisplatin and ethyl acetate extract of Vernonia amygdalina Delile induces cell cycle arrest and apoptosis on PANC-1 cells via PI3K/mTOR [Combinación de cisplatino y extracto de acetato de etilo de Vernonia amygdalina Delile induce la detención del … Continue reading Cisplatin-Vernonia amygdalina combination on pancreatic cancer

J. Pharm. Pharmacogn. Res., vol. 12, no. 5, pp. 870-880, Sep-Oct 2024.

DOI: https://doi.org/10.56499/jppres23.1748_12.5.870

Original Article

Combination of cisplatin and ethyl acetate extract of Vernonia amygdalina Delile induces cell cycle arrest and apoptosis on PANC-1 cells via PI3K/mTOR

[Combinación de cisplatino y extracto de acetato de etilo de Vernonia amygdalina Delile induce la detención del ciclo celular y la apoptosis en células PANC-1 vía PI3K/mTOR]

Poppy Anjelisa Zaitun Hasibuan1*, Jane Melita Keliat2, Muhammad Fauzan Lubis3

1Department of Pharmacology, Faculty of Pharmacy, Universitas Sumatera Utara, Medan, 20155, Indonesia.

2Department of Pharmaceutical and Food Analysis, Faculty of Vocational, Universitas Sumatera Utara, Medan, 20155, Indonesia.

3Department of Pharmaceutical Biology, Faculty of Pharmacy, Universitas Sumatera Utara, Medan, 20155, Indonesia.

*E-mail: poppyanjelisa@usu.ac.id

Abstract

Context: Vernonia amygdalina Delile (VAD) is known as a potential plant with a wide variety of medicinal properties, including anticancer.

Aims: To evaluate the combination effect of VAD extract with cisplatin (CIS) against PANC-1 cells, focusing on cell cycle arrest and apoptosis activities.

Methods: This study is an experimental study using PANC-1 cells as objects. The phytochemical compounds were analyzed with LC-MS/MS. The cytotoxic activity of extracts and their combinations was determined using the MTT assay method in the PANC-1 cell line. Apoptosis, cell cycle arrest, and PI3K/mTOR profiles were analyzed with flow cytometry. Immunocytochemistry was used to determine Bcl-2, Cyclin D1, and p53 expression.

Results: The phytochemicals found were five compounds with retention times of 8.96, 9.85, 10.52, 12.59, and 10.36. The results of the MTT assay showed that the IC50 values of extract and CIS were 21.83 ± 0.46 µg/mL and 3.02 ± 0.44 µg/mL, respectively. The combination index (CI) value of extract and CIS had a synergistic effect. Combination extract and CIS induced early and late apoptosis, inhibited cell cycle progression on the G1 phase, inhibited Bcl-2 and cyclin D1 expression, induced p53 expression, and inhibited PI3K and mTOR expression.

Conclusions: The combination of extract and CIS showed anticancer activity against PANC-1 cells through induction of apoptosis and cell cycle arrest via inhibition of PI3K and mTOR expressions.

Keywords: apoptosis; cell cycle arrest; cisplatin; herbal medicine; pancreatic cancer.

PDF Download

Resumen

Contexto: Vernonia amygdalina Delile (VAD) es conocida como una planta potencial con una amplia variedad de propiedades medicinales, incluyendo anticancerígenas.

Objetivos: Evaluar el efecto combinado del extracto de VAD con cisplatino (CIS) contra las células PANC-1, centrándose en las actividades de detención del ciclo celular y apoptosis.

Métodos: Este estudio es un estudio experimental utilizando células PANC-1 como objetos. El análisis de los compuestos fitoquímicos se llevó a cabo con LC-MS/MS. La actividad citotóxica del extracto y sus combinaciones se determinó mediante el método de ensayo MTT en la línea celular PANC-1. La apoptosis, la detención del ciclo celular y los perfiles PI3K/mTOR se analizaron con citometría de flujo. Se utilizó inmunocitoquímica para determinar la expresión de Bcl-2, ciclina D1 y p53.

Resultados: Los fitoquímicos hallados fueron cinco compuestos con tiempos de retención de 8,96, 9,85, 10,52, 12,59 y 10,36. Los resultados del ensayo MTT mostraron que los valores de IC50 del extracto y del CIS fueron 21,83 ± 0,46 µg/mL y 3,02 ± 0,44 µg/mL, respectivamente. El valor del índice de combinación (IC) del extracto y el CIS tuvo un efecto sinérgico. La combinación de extracto y CIS indujo la apoptosis temprana y tardía, inhibió la progresión del ciclo celular en la fase G1, inhibió la expresión de Bcl-2 y ciclina D1, indujo la expresión de p53 e inhibió la expresión de PI3K y mTOR.

Conclusiones: La combinación de extracto y CIS mostró actividad anticancerígena contra células PANC-1 a través de la inducción de apoptosis y detención del ciclo celular vía inhibición de las expresiones PI3K y mTOR.

Palabras Clave: apoptosis; detención del ciclo celular; cisplatino; fitoterapia; cáncer de páncreas.

PDF Download

 

 
 
Citation Format: Hasibuan PAZ, Keliat JM, Lubis MF (2024) Combination of cisplatin and ethyl acetate extract of Vernonia amygdalina Delile induces cell cycle arrest and apoptosis on PANC-1 cells via PI3K/mTOR. J Pharm Pharmacogn Res 12(5): 870–880. https://doi.org/10.56499/jppres23.1748_12.5.870
References

Aamazadeh F, Ostadrahimi A, Rahbar Saadat Y, Barar J (2020) Bitter apricot ethanolic extract induces apoptosis through increasing expression of Bax/Bcl-2 ratio and caspase-3 in PANC-1 pancreatic cancer cells. Mol Biol Rep 47: 1895–1904. https://doi.org/10.1007/s11033-020-05286-w

Akinleye A, Avvaru P, Furqan M, Song Y, Liu D (2013) Phosphatidylinositol 3-kinase (PI3K) inhibitors as cancer therapeutics. J Hermatol Oncol 6: 88. https://doi.org/10.1186/1756-8722-6-88

Aldossary SA (2019) Review on pharmacology of cisplatin: Clinical use, toxicity and mechanism of resistance of cisplatin. Biomed Pharmacol J 12: 7–15. https://doi.org/10.13005/bpj/1608

Barabas K, Milner R, Lurie D, Adin C (2008) Cisplatin: a review of toxicities and therapeutic applications Vet Comp Oncol 6: 1–18. https://doi.org/10.1111/j.1476-5829.2007.00142.x

Chao Y, Wu CY, Wang JP, Lee RC, Lee WP, Li CP (2013) A randomized controlled trial of gemcitabine plus cisplatin versus gemcitabine alone in the treatment of metastatic pancreatic cancer. Cancer Chemother Pharmacol 72: 637–642. https://doi.org/10.1007/s00280-013-2239-1

Chen XL, Cheng QY, She MR, Wang Q, Huang XH, Cao LQ, Fu XH, Chen JS (2010) Expression of sonic hedgehog signaling components in hepatocellular carcinoma and cyclopamine-induced apoptosis through Bcl-2 downregulation in vitro. Arch Med Res 41: 315–323. https://doi.org/10.1016/j.arcmed.2010.06.003

Dalimunthe A, Hasibuan PAZ, Satria D (2017) Cell cycle arrest activity of Litsea cubeba Lour: Heartwood and fruit extracts against T47D breast cancer cells. Asian J Pharm Clin Res 10: 404–406. https://doi.org/10.22159/ajpcr.2017.v10i11.20204

Dong Q, Ling B, Gao B, Maley J, Sammynaiken R, Yang J (2014) Hedyotis diffusa water extract diminished the cytotoxic effects of chemotherapy drugs against human breast cancer MCF7 cells. Nat Prod Commun 9: 699–700. https://doi.org/10.1177/1934578x1400900529

Eldhose B, Gunawan M, Rahman M, Latha MS, Notario V (2014) Plumbagin reduces human colon cancer cell survival by inducing cell cycle arrest and mitochondria-mediated apoptosis. Int J Oncol 45: 1913–1920. https://doi.org/10.3892/ijo.2014.2592

Ergun Y, Ozdemir NY, Guner EK, Esin E, Sendur MA, Koksoy EB, Demirci NS, Eren T, Dede I, Sezer A, Engin H, Oksuzoglu B, Yalcin B, Utkan G, Zengin N, Urun Y (2018) Comparison of gemcitabine monotherapy with gemcitabine and cisplatin combination in metastatic pancreatic cancer: a retrospective analysis. J BUON 23: 116–121. https://pubmed.ncbi.nlm.nih.gov/30722120/

Fachrunisa D, Hasibuan PAZ, Harahap U (2019) Cell cycle inhibition and apoptotic induction of Vernonia amygdalina Del. leaves extract on MCF-7 cell line. Open Access Maced J Med Sci 7: 3807–3810. https://doi.org/10.3889/oamjms.2019.509

Febriansah R, Putri DD, Sarmoko, Nurulita NA, Meiyanto E, Nugroho AE (2014) Hesperidin as a preventive resistance agent in MCF-7 breast cancer cells line resistance to doxorubicin. Asian Pac J Trop Biomed 4: 228–233. https://doi.org/10.1016/S2221-1691(14)60236-7

Haryanti S, Pramono S, Murwanti R, Meiyanto E (2016) The synergistic effect of doxorubicin and ethanolic extracts of Caesalpinia sappan L. wood and Ficus septica Burm. f. leaves on viability, cell cycle progression, and apoptosis induction of MCF­7 cells. Indones J Biotech 21: 29–37. https://doi.org/10.22146/ijbiotech.26105

Hasibuan PAZ, Chrestella J, Satria D (2015) Combination effect of ethylacetate extracts of Plectranthus amboinicus (Lour.) Spreng. with doxorubicin againts T47D breast cancer cells. Int J Pharm Pharm Sci 7: 156–159.

Hasibuan PAZ, Harahap U, Sitorus P, Lubis MF, Satria D (2021) In-silico analysis of vernonioside D and vernonioside E from Vernonia amygdalina Delile. leaves as inhibitor of epidermal growth factor receptor (EGFR) and mammalian target of rapamycin (mTOR). Rasayan J Chem 14: 1539–1543. https://doi.org/10.31788/RJC.2021.1436092

Hasibuan PAZ, Harahap U, Sitorus P, Satria D (2020b) The anticancer activities of Vernonia amygdalina Delile. leaves on 4T1 breast cancer cells through phosphoinositide 3-kinase (PI3K) pathway. Heliyon 6: e04449. https://doi.org/10.1016/j.heliyon.2020.e04449

Hasibuan PAZ, Keliat JM, Lubis MF, Nasution A (2024) The ethyl acetate extract of Vernonia amygdalina leaf ameliorates gemcitabine effect against migration and invasion of PANC-1 cells via down-regulation the VEGF, COX2, and RAS/MEK pathways. Saudi Pharm J 32: 101872. https://doi.org/10.1016/j.jsps.2023.101872

Hasibuan PAZ, Lubis MF, Keliat JM, Azizah N (2023) Cytotoxic test combination of ethyl acetate extract africant leaves (Vernonia amygdalina Delile) and gemcitabine on PANC-1 cells. AIP Conference Proceedings 2626: 030004. https://doi.org/10.1063/5.0149754

Hasibuan PAZ, Munir D, Pertiwi D, Satria D, Lubis MF (2020a) Flavonoids constituent analysis and cell cycle inhibition activity of ethylacetate extract of Vernonia amygdalina Delile. leaves on lung cancer cell line. Rasayan J Chem 13: 2577–2581. https://doi.org/10.31788/RJC.2020.1345625

Hasibuan PAZ, Sumaiyah S (2019) The anti-proliferative and pro-apoptotic properties of ethanol Plectranthus amboinicus (Lour.) Spreng. leaves ethanolic extract nanoparticles on T47D cell lines. Asian Pac J Cancer Prev 20: 897–901. https://doi.org/10.31557/APJCP.2019.20.3.897

Heinemann V, Wilke H, Mergenthaler HG, Clemens M, König H, Illiger HJ, Arning M, Schalhorn A, Possinger K, Fink U (2000) Gemcitabine and cisplatin in the treatment of advanced or metastatic pancreatic cancer. Ann Oncol 11: 1399–1403. https://doi.org/10.1023/a:1026595525977

Housman G, Byler S, Heerboth S, Lapinska K, Longacre M, Snyder N, Sarkar S (2014) Drug resistance in cancer: An overview. Cancers (Basel) 6: 1769–1792. https://doi.org/10.3390/cancers6031769

Illian DN, Hasibuan PAZ, Sumardi S, Nuryawan A, Wati R, Basyuni M (2019) Anticancer activity of polyisoprenoids from Avicennia alba Blume. in WiDr cells. Iran J Pharm Res 18: 1477–1487. https://doi.org/10.22037/ijpr.2019.1100719

Istiqomah, Meighina A, Hasibuan PAZ, Sumaiyah S, Yusraini E, Oku H, Basyuni M (2020) Anticancer effects of polyisoprenoid from Nypa fruticans leaves by controlling expression of P53, EGFR, PI3K, AKT1, and MTOR genes in colon cancer (WiDr) cells. Nat Prod Commun 15: 1–8. https://doi.org/10.1177/1934578X20918412

Kabała-Dzik A, Rzepecka-Stojko A, Kubina R, Iriti M, Wojtyczka RD, Buszman E, Stojko J (2018) Flavonoids, bioactive components of propolis, exhibit cytotoxic activity and induce cell cycle arrest and apoptosis in human breast cancer cells MDA-MB-231 and MCF-7 - a comparative study. Cell Mol Biol (Noisy-le-grand) 64: 1–10. https://pubmed.ncbi.nlm.nih.gov/29981677/

Kapadia GJ, Rao GS, Ramachandran C, Iida A, Suzuki N, Tokuda H (2013) Synergistic cytotoxicity of red beetroot (Beta vulgaris L.) extract with doxorubicin in human pancreatic, breast and prostate cancer cell lines. J Complement Integr Med 10: 113–122. https://doi.org/10.1515/jcim-2013-0007

Karanikas M, Esempidis A, Chasan ZT, Deftereou T, Antonopoulou M, Bozali F, Amarantidis K, Man YG (2016) Pancreatic cancer from molecular pathways to treatment opinion. J Cancer 7: 1328–1339. https://doi.org/10.7150/jca.15419

Kim M, Kim YS, Kim KM, Ko HC, Kim SJ, Kim JH, Kim Y (2014) Combination of Sasa quelpaertensis Nakai leaf extract and cisplatin suppresses the cancer stemness and invasion of human lung cancer cells. Integr Cancer Ther 13: 529–540. https://doi.org/10.1177/1534735414534462

Kong F, Liu X, Zhou Y, Hou X, He J, Li Q, Miao X, Yang L (2020) Downregulation of METTL14 increases apoptosis and autophagy induced by cisplatin in pancreatic cancer cells. Int J Biochem Cell Biol 122: 105731. https://doi.org/10.1016/j.biocel.2020.105731

Kotawong K, Chaijaroenkul W, Muhamad P, Na-Bangchang K (2018) Cytotoxic activities and effects of atractylodin and β-eudesmol on the cell cycle arrest and apoptosis on cholangiocarcinoma cell line. J Pharmacol Sci 136: 51–56. https://doi.org/10.1016/j.jphs.2017.09.033

Li Y, Wang T, Sun Y, Huang T, Li C, Fu Y, Li Y, Li C (2019) p53-Mediated PI3K/AKT/mTOR Pathway Played a Role in PtoxDpt-Induced EMT Inhibition in Liver Cancer Cell Lines. Oxid Med Cell Longev 2019: 2531493. https://doi.org/10.1155/2019/2531493

Lubis MF, Hasibuan PAZ, Harahap U (2019) Phytochemicals screening and cell cycle arrest activity of n-Hexane extract of Vernonia amygdalina Delile leaves against pancreatic cancer cell line. Asian J Pharm Res Dev 7: 12–16. https://doi.org/10.22270/ajprd.v7i4.533

Lubis MF, Hasibuan PAZ, Harahap U, Satria D, Syahputra H, Muhammad M, Astyka R (2022) The molecular approach of natural products as pancreatic cancer treatment: a review. Rasayan J Chem 15: 1362–1377. https://doi.org/10.31788/RJC.2022.1526765

Mao Y, Xi L, Li Q, Cai Z, Lai Y, Zhang X, Yu C (2016) Regulation of cell apoptosis and proliferation in pancreatic cancer through PI3K/Akt pathway via Polo-like kinase 1. Oncol Rep 36: 49–56. https://doi.org/10.3892/or.2016.4820

Nikolaou M, Pavlopoulou A, Georgakilas AG, Kyrodimos E (2018) The challenge of drug resistance in cancer treatment: A current overview. Clin Exp Metastasis 35: 309–318. https://doi.org/10.1007/s10585-018-9903-0

Ohno I, Eibl G, Odinokova I, Edderkaoui M, Damoiseaux RD, Yazbec M, Abrol R, Goddard WA 3rd, Yokosuka O, Pandol SJ, Gukovskaya AS (2010) Rottlerin stimulates apoptosis in pancreatic cancer cells through interactions with proteins of the Bcl-2 family. Am J Physiol Gastrointest Liver Physiol 298: G63–G73. https://doi.org/10.1152/ajpgi.00257.2009

Ruzzolini J, Peppicelli S, Andreucci E, Bianchini F, Scardigli A, Romani A, la Marca G, Nediani C, Calorini L (2018) Oleuropein, the main polyphenol of Olea europaea leaf extract, has an anti-cancer effect on human braf melanoma cells and potentiates the cytotoxicity of current chemotherapies. Nutrients 10: 1950. https://doi.org/10.3390/nu10121950

Satria D, Silalahi J, Haro G, Ilyas S, Hasibuan PAZ (2019) Chemical analysis and cytotoxic activity of n-hexane fraction of Zanthoxylum acanthopodium DC. fruits. Rasayan J Chem 12: 803–808. https://doi.org/10.31788/RJC.2019.1225180

Sutejo IK, Putri H, Handayani S, Jenie RI, Meiyanto E (2019) In vitro study of the combination of doxorubicin, Curcuma xanthorrhiza, Brucea javanica, and Ficus septica as a potential novel therapy for metastatic breast cancer. Indones J Pharm 30: 15–24. https://doi.org/10.14499/indonesianjpharm30iss1pp15

Syari DM, Rosidah R, Hasibuan PAZ, Haro G, Satria D (2019) Evaluation of cytotoxic activity alkaloid fractions of Zanthoxylum acanthopodium DC. fruits. Open Access Maced J Med Sci 7: 3745–3747. https://doi.org/10.3889/oamjms.2019.495

Szliszka E, Czuba ZP, Domino M, Mazur B, Zydowicz G, Krol W (2009) Ethanolic extract of propolis (EEP) enhances the apoptosis- inducing potential of TRAIL in cancer cells. Molecules 14: 738–754. https://doi.org/10.3390/molecules14020738

Teodoro AJ, Oliveira FL, Martins NB, Maia Gde A, Martucci RB, Borojevic R (2012) Effect of lycopene on cell viability and cell cycle progression in human cancer cell lines. Cancer Cell Int 12: 36. https://doi.org/10.1186/1475-2867-12-36

Tine Y, Yang Y, Renucci F, Costa J, Wélé A, Paolini J (2017) LC-MS/MS analysis of flavonoid compounds from Zanthoxylum zanthoxyloides extracts and their antioxidant activities. Nat Prod Commun 12: 1865–1868. https://doi.org/10.1177/1934578x1701201213

Topçu G, Ayral MN, Aydin A, Gören AC, Chai HB, Pezzuto JM (2001) Triterpenoids of the roots of Lavandula stoechas ssp. stoechas. Pharmazie 56: 892–895. https://pubmed.ncbi.nlm.nih.gov/11817178/

Tseng CY, Lin CH, Wu LY, Wang JS, Chung MC, Chang JF, Chao MW (2016) Potential combinational anti-cancer therapy in non-small cell lung cancer with traditional Chinese medicine Sun-Bai-Pi extract and cisplatin. PLoS One 11: e0155469. https://doi.org/10.1371/journal.pone.0155469

Tuasha N, Escobar Z, Seifu D, Gadisa E, Petros B, Sterner O, Oredsson S (2022) Cytotoxic and other bioactivities of a novel and known sesquiterpene lactones isolated from Vernonia leopoldi (Sch. Bip. ex Walp.) Vatke in breast cancer cell lines. Toxicol Rep 9: 382–392. https://doi.org/10.1016/j.toxrep.2022.02.011

Uroz M, Wistorf S, Serra-Picamal X, Conte V, Sales-Pardo M, Roca-Cusachs P, Guimerà R, Trepat X (2018) Regulation of cell cycle progression by cell-cell and cell-matrix forces. Nat Cell Biol 20: 646–654. https://doi.org/10.1038/s41556-018-0107-2

Wang X, Wang X, Xu Y, Yan M, Li W, Chen J, Chen T (2020) Effect of nicastrin on hepatocellular carcinoma proliferation and apoptosis through PI3K/AKT signalling pathway modulation. Cancer Cell Int 20: 91. https://doi.org/10.1186/s12935-020-01172-4

Werner J, Combs SE, Springfeld C, Hartwig W, Hackert T, Büchler MW (2013) Advanced-stage pancreatic cancer: therapy options. Nat Rev Clin Oncol 10: 323–333. https://doi.org/10.1038/nrclinonc.2013.66

Williams GH, Stoeber K (2012) The cell cycle and cancer. J Pathol 226: 352–364. https://doi.org/10.1002/path.3022

Xiao Z, Jiang Y, Wang CQ, Hu SS, Huang XR, Chen XF, Huang J, Shan LJ, Tang YH, Wang YH, Gong QH, Feng JH, Xiao X, Li XF (2020) Clinical efficacy and safety of aidi injection combination with vinorelbine and cisplatin for advanced non-small-cell lung carcinoma: A systematic review and meta-analysis of 54 randomized controlled trials. Pharmacol Res 153: 104637. https://doi.org/10.1016/j.phrs.2020.104637

Zhang X, Ren J, Cheng X, Jin H, Zhang W (2015) One new unusual sesterterpenoid and four new sesquiterpene dimers from Inula britannica. RSC Advances 5: 1979–1982. https://doi.org/10.1039/C4RA11171K

© 2024 Journal of Pharmacy & Pharmacognosy Research

Anti-dormant mycobacterial of marine-derived fungi
J. Pharm. Pharmacogn. Res., vol. 13, no. 1, pp. 16-26, Jan-Feb 2025. DOI: https://doi.org/10.56499/jppres24.1953_13.1.16 Original Article Activity of ethyl acetate extracts of marine-derived fungi against active and hypoxia-induced dormant Mycobacterium [Actividad de extractos de acetato de etilo de hongos de origen marino contra Mycobacterium latente activa e inducida por hipoxia] Muhammad Azhari1, Atik Pereztia Litanjuasari1, … Continue reading Anti-dormant mycobacterial of marine-derived fungi
Rift Valley fever virus RdRp inhibition by RNA polymerase inhibitors
J. Pharm. Pharmacogn. Res., vol. 13, no. 1, pp. 1-15, Jan-Feb 2025. DOI: https://doi.org/10.56499/jppres24.1967_13.1.1 Original Article In silico study of RNA polymerase inhibitor drugs for Rift Valley fever virus using RdRp protein as the target [Estudio in silico de fármacos inhibidores de la ARN polimerasa para el virus de la fiebre del valle del Rift … Continue reading Rift Valley fever virus RdRp inhibition by RNA polymerase inhibitors
Probable interaction between levothyroxine and Thymus vulgaris
J. Pharm. Pharmacogn. Res., vol. 12, no. 6, pp. 1196-1198, Nov-Dec 2024. DOI: https://doi.org/10.56499/jppres24.2008_12.6.1196 Case Report Probable interaction between levothyroxine sodium and thyme (Thymus vulgaris), about a case report [Interacción probable entre levotiroxina sódica y tomillo (Thymus vulgaris), sobre un reporte de caso] Nassima Elyebdri1,2*, Sihem Baba Ahmed1, Nessrine Abourejal1, Lotfi Loudjedi3, Assia Bououden3, Nour … Continue reading Probable interaction between levothyroxine and Thymus vulgaris

© 2013-2020 by the authors; licensee JPPRes, Antofagasta, Chile. This journal is an open-access journal distributed under the terms and conditions of the Creative Commons Attribution license-Non Commercial 4.0 international. The content on this site is intended for health professionals. If you are not a health professional, please talk to your doctor about any doubts or concerns regarding your health

Made with ♥ by AVAGAX Studio