Author(s): Liesl Maria Fernandese Mendonca, Arun Bhimrao Joshi, Anant Bhandarkar, Himanshu Joshi

Email(s): lieslpharma@gmail.com , joshiarun0264@gmail.com , anantpharm@gmail.com , 2hjoshi@gmail.com

DOI: 10.52711/0974-360X.2023.00675   

Address: Liesl Maria Fernandese Mendonca1*, Arun Bhimrao Joshi2, Anant Bhandarkar2, Himanshu Joshi3
1Department of Pharmacology, Goa College of Pharmacy, 18th June Road, Panaji, Goa 403001 India.
2Department of Pharmacognosy and Phytochemistry, Goa College of Pharmacy, 18th June Road, Panaji, Goa, 403001 India.
3Department of Pharmacology, College of Pharmacy, Graphic Era Hill University, Bhimtal campus, Uttarakhand 263156 India.
*Corresponding Author

Published In:   Volume - 16,      Issue - 9,     Year - 2023


ABSTRACT:
Objective: According to the World Health Organisation, breast cancer is presently the most common cancer diagnosed in women globally. Polyphenolic compounds act as antioxidants, improve health, and reduce risk and proliferation of various types of cancers. Hybanthus enneaspermus Linn. is a beneficial medicinal plant, reported to possess antimicrobial, antioxidant, anti-inflammatory, neuroprotective, cardioprotective, and nephroprotective properties etc. Methods: The current study involved the evaluation the antioxidant, antiproliferative, apoptotic and cell cycle arrest potential of the ethanolic leaf extract of Hybathus enneaspermus Linn. (EEHE), its toluene soluble, toluene insoluble, ethyl acetate and methanol soluble biofractions viz. TFHE, ITHE, EFHE, and MFHE to combat breast cancer. In vitro antioxidant activities were evaluated using DPPH, Hydrogen peroxide, Nitric oxide and ABTS free radical scavenging assays. In vitro antiproliferative activity against MCF-7 cells was assayed using the Sulforhodamine method, while apoptosis and cell cycle assays were analysed by flow cytometry. Results: MFHE exhibited significant in vitro antioxidant activity with IC50 values of 21.10±0.39 µg/mL and 25.99±4.66µg/mL, when compared against standard ascorbic acid with IC50 values of 11.19±1.09 µg/mL and 9.30±0.26µg/mL in DPPH and nitric oxide assays respectively. EFHE displayed substantial antioxidant potential in ABTS and hydrogen peroxide assays with IC50 values of 40.38±0.88µg/mL and 99.11± 13.59µg/mL, while ITHE showed considerable activity with IC50 < 100µg/mL in DPPH, nitric oxide and ABTS assays. TFHE demonstrated significant antiproliferative activity by sulforhodamine assay, with GI50 value of 10.22 6.72µg/mL, while EEHE and ITHE showed substantial activity with GI50 values of 41.42±3.74µg/mL and 64.37±7.07µg/mL respectively, as against the standard drug Adriamycin (GI50 < 10µg/mL) used. In the apoptosis assay, ITHE showed 11.31±0.82% cells in late apoptosis and 34.48±1.57 % cells in necrosis as compared to standard Adriamycin indicated 13.67±1.02 % cells in late apoptosis and 8.58±0.65 % cells in necrosis. In cell cycle analysis, ITHE displayed significant apoptotic activity with 20.15±1.37 % cells in SubG1 phase and 13.99±1.65 % cells arrested in G2-M phase as compared to the control. Conclusion: The study thus revealed that MFHE, EFHE and ITHE biofractions showed significant antioxidant activities, while EEHE, TFHE and ITHE exhibited substantial antiproliferative activity against mammary cancer cells. Additionally, ITHE induced remarkable apoptotic activity and cell cycle arrest in the MCF-7 cells. The therapeutic benefits may be credited to the bioactive constituents present in the ITHE fraction viz. polyphenolics, flavonoids etc.; however, the molecular mechanisms may need to be evaluated further.


Cite this article:
Liesl Maria Fernandese Mendonca, Arun Bhimrao Joshi, Anant Bhandarkar, Himanshu Joshi. Antioxidant, Antiproliferative, Pro-apoptotic and cell cycle arrest properties of crude extract and biofractions of Hybanthus enneaspermus Linn. to combat breast cancer. Research Journal of Pharmacy and Technology 2023; 16(9):4127-4. doi: 10.52711/0974-360X.2023.00675

Cite(Electronic):
Liesl Maria Fernandese Mendonca, Arun Bhimrao Joshi, Anant Bhandarkar, Himanshu Joshi. Antioxidant, Antiproliferative, Pro-apoptotic and cell cycle arrest properties of crude extract and biofractions of Hybanthus enneaspermus Linn. to combat breast cancer. Research Journal of Pharmacy and Technology 2023; 16(9):4127-4. doi: 10.52711/0974-360X.2023.00675   Available on: https://www.rjptonline.org/AbstractView.aspx?PID=2023-16-9-19


REFERENCES:
1.    Wilkinson L, Gathani T. Understanding breast cancer as a global health concern. Bri. J. Radiol. 2022;95(1130):20211033. doi:10.1259/bjr.20211033
2.    Yadav AR, Mohite SK. Anticancer activity of Psidium guajava leaf extracts on breast cancer cell line. Res.  J. Pharm. Dos. Forms Techol. 2020;12(4):298-300. doi:10.5958/0975-4377.2020.00049.X
3.    Abotaleb M, Samuel SM, Varghese E, Varghese S, Kubatka P, Liskova A, Büsselberg D. Flavonoids in cancer and apoptosis. Cancers. 2018;11(1):28. doi:10.3390/cancers11010028
4.    Greenwell M, Rahman PK. Medicinal plants: their use in anticancer treatment. Int. J. Pharm. Sci. Res. 2015;6(10):4103. doi: 10.13040/IJPSR.0975-8232.6(10).4103-12
5.    Seena H, Kannappan N, Kumar PM. In vitro Antioxidant and Anticancer activity of methanolic extract of Alangium salvifolium subsp. hexapetalum (Wangerin). Research J. Pharm. and Tech. 2020;13(8):3715-9. doi: 10.5958/0974-360X.2020.00657.5
6.    Balabhaskar R, Vijayalakshmi K. Evaluation of anticancer activity of ethanol extract of Bauhinia tomentosa linn. on A549, human lung carcinoma cell lines. Res. J. Pharm. Technol. 2019;12(6):2748-52. doi: 10.5958/0974-360X.2019.00460.8
7.    Gupta P, Khan MY, Verma VK, Pathak A. Beating Cancer with Natural Plant Sources. Asian J. Pharm. Tech. 2013;3(2):39-44.
8.    Vuda M, D'Souza R, Upadhya S, Kumar V, Rao N, Kumar V, Boillat C, Mungli P. Hepatoprotective and antioxidant activity of aqueous extract of Hybanthus enneaspermus against CCl4-induced liver injury in rats. Exp. Toxicol Pathol. 2012;64(7-8):855-9. doi: 10.1016/j.etp.2011.03.006
9.    Patel DK, Kumar R, Prasad SK, Sairam K, Hemalatha S. Antidiabetic and in vitro antioxidant potential of Hybanthus enneaspermus (Linn) F. Muell in streptozotocin–induced diabetic rats. Asian Pac. J. Trop. Biomed. 2011;1(4):316-22. doi: 10.1016/S2221-1691(11)60051-8
10.    Rajsekhar PB, Bharani RA, Angel KJ, Ramachandran M, Rajsekhar SP. Hybanthus enneaspermus (L) F. Muell: A phytopharmacological review on herbal medicine. J. Chem. Pharm. Res. 2016;8(1):351-5.
11.    Patankar P, Biradar SR. Biochemical Studies in Hybanthus enneaspermus (L.) F. Muell. Think India Journal. 2019;22(31):490-4.
12.    Patel DK, Kumar R, Laloo D, Hemalatha S. Evaluation of phytochemical and antioxidant activities of the different fractions of Hybanthus enneaspermus (Linn.) F. Muell.(Violaceae). Asian Pac. J. Trop. Med. 2011;4(5):391-6. doi: 10.1016/S1995-7645(11)60110-7.
13.    Sundaram S, Radhakrishnan A, Kanniappan GV, Bhaskaran SK, Palanisamy CP, Kannappan P. Comparative Study on Antioxidant Activity of Crude and Alkaloid Extracts of Hybanthus    enneaspermus (Linn) F. Mull. Anal. Chem. Lett. 2015;5(5):291-9. doi: 10.1080/22297928.2015.1135076
14.    Khandelwal, K. R. 2010. Practical Pharmacognosy. Techniques and Experiments. 20th Edn. Nirali Prakashan.
15.    Trease G. E. & Evans W. C.1989. Trease and Evan’s Textbook of Pharmacognosy. 13th Edn. Cambridge University Press, London. 546.
16.    Chakraborty P, Sharma S, Chakraborty S, Siddapurand A, Abraham J. Cytotoxicity and Antimicrobial Activity of Ipomoea batatas. Res. J. Pharm. Technol. 2018;11(7):2741-6. doi:  10.5958/0974-360X.2018.00506.1
17.    Chanda S, Dave R. In vitro models for antioxidant activity evaluation and some medicinal plants possessing antioxidant properties: An overview. Afr. J. Microbiol. Res. 2009;3(13):981-96.
18.    Ghagane SC, Puranik SI, Kumbar VM, Nerli RB, Jalalpure SS, Hiremath MB, Neelagund S, Aladakatti R. In vitro antioxidant and anticancer activity of Leea indica leaf extracts on human prostate cancer cell lines. Integr. Med. Res. 2017;6(1):79-87. doi: 10.1016/j.imr.2017.01.004
19.    Chen Z, Bertin R, Froldi G. EC50 estimation of antioxidant activity in DPPH assay using several statistical programs. Food Chem. 2013;138(1):414-20. doi: 10.1016/j.foodchem.2012.11.001
20.    Raju DC, Victoria TD, Biji N, Nikitha G. Evaluation of antioxidant potential of ethanolic extract of Centella asiatica L. Res. J. Pharm. Technol. 2015;8(9):1289-93. doi: 10.5958/0974-360X.2015.00234.6
21.    Ruch RJ, Cheng SJ, Klaunig JE. Prevention of cytotoxicity and inhibition of intercellular communication by antioxidant catechins isolated from Chinese green tea. Carcinogenesis. 1989;10(6):1003-8. doi: 10.1093/carcin/10.6.1003
22.    Boora F, Chirisa E, Mukanganyama S. Evaluation of nitrite radical scavenging properties of selected Zimbabwean plant extracts and their phytoconstituents. J. Food Process. 2014;2014. doi: 10.1155/2014/918018
23.    Krishnaveni M, Santhoshkumar J. Secondary metabolite, antioxidant, phyto nutrient assay of essential oil from dry Coriandrum sativum seed black variety. Res. J. Pharm. Technol. 2016;9(7):853. doi: 0.5958/0974-360X.2016.00161.X
24.    Praveen D, Chowdary PR, Thanmayi G, Poojitha G, Aanandhi MV. Antioxidant and analgesic activity of leaf extracts of Artocarpus heterophyllus. Res. J. Pharm. Technol. 2016;9(3):257-61. doi: 10.5958/0974-360X.2016.00047.0
25.    Vaghela JS, Sisodia SS. In vitro antioxidant activity of Terminalia chebula fruit extracts. Res. J. Pharm. Technol. 2011;4(12):1835-43.
26.    Prior RL, Wu X, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric. Food Chem. 2005;53(10):4290-302. doi: 10.1021/jf0502698
27.    Salama ZA, Aboul-Enein AM, Gaafar AA, Asker MS, Aly HF, Ahmed HA. In-vitro antioxidant, antimicrobial and anticancer activities of banana leaves (Musa acuminata) and olive leaves (Olea europaea L.) as by-products. Res. J. Pharm. Technol. 2020;13(2):687-96. doi: 10.5958/0974-360X.2020.00132.8
28.    Vichai V, Kirtikara K. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat. protoc. 2006;1(3):1112-6. doi:10.1038/nprot.2006.179
29.    Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR. New colorimetric cytotoxicity assay for anticancer-drug screening. J. Nat. Cancer Inst. 1990; 82(13):1107-12.  doi:10.1093/jnci/82.13.1107
30.    Mendonça LM, Joshi AB, Bhandarkar AV, Joshi H. Evaluation of antioxidant property and anticancer prospective of the leaf extract and biofractions of Bauhinia foveolata Dalzell–A native of the Indian Western Ghats. Int. J. Res Pharm Sci. 2021;12(3):1886-94. doi:10.26452/ijrps.v12i3.4789
31.    Pozarowski P, Grabarek J, Darzynkiewicz Z. Flow cytometry of apoptosis. Curr. Protoc. Cell Biol. 2003;18.8.1-33.    doi:10.1002/0471143030.cb1808s21
32.    Tilekar K, Upadhyay N, Jänsch N, Schweipert M, Mrowka P, Meyer-Almes FJ, Ramaa CS. Discovery of 5-naphthylidene-2, 4-thiazolidinedione derivatives as selective HDAC8 inhibitors and evaluation of their cytotoxic effects in leukemic cell lines. Bioorg. chem. 2020;95:103522.  doi:10.1016/j.bioorg.2019.103522
33.    Fridlender M, Kapulnik Y, Koltai H. Plant derived substances with anti-cancer activity: from folklore to practice. Front. Plant Sci. 2015;6:799.  doi: 10.3389/fpls.2015.00799
34.    Banu S, Ramakrishnaiah TN. Screening of Garcinia cambogia for in-vitro anti-cancerous activity against colon adenocarcinoma (Caco-2) cell line. Res. J. Pharmacogn. Phytochem. 2018;10(4):272-6. doi: 10.5958/0975-4385.2018.00043.2
35.    Ramos S. Effects of dietary flavonoids on apoptotic pathways related to cancer chemoprevention. The J. Nutr. Biochem. 2007;18(7):427-42. doi: 10.1016/j.jnutbio.2006.11.004
36.    Merlin NJ, Parthasarathy V, Manavalan R, Devi P, Meera R. Apoptosis Significance and Molecular Mechanisms-A Review. Asian J. Res. Chem. 2009;2(4):369-75
37.    Fatma F, Kumar A. The Cell Cycle, Cyclins, Checkpoints and Cancer. Asian J. Res. Pharm. Sci. 2021;11(2):175-83. doi: 10.52711/2231-5659.2021-11-2-14
38.    Rangasamy P, Hansiya VS, Maheswari PU, Suman T, Geetha N. Phytochemical analysis and evaluation of in vitro antioxidant and anti-urolithiatic potential of various fractions of Clitoria ternatea L. Blue flowered leaves. Asian J. Pharm. Ana. 2019;9(2):67-76. doi: 10.5958/2231-5675.2019.00014.0
39.    Awati SS, Singh SK, Wadkar KA. In vitro Antioxidant potential and Anticancer activity of Ceratophyllum demersum Linn. extracts on HT-29 human colon cancer cell line. Res. J. Pharm. Technol. 2021;14(1):28-36. doi: 10.5958/0974-360X.2021.00006.8
40.    Islam NN, Naine SJ, Jameel S, Khora SS, Kumar V, Mohanasrinivasan V, Devi CS, Panneerselvam A. Cytotoxic Property of Cocos nucifera shell Extracts on HeLa Cells. Res. J. Pharm. Technol. 2014;7(5):521-5.
41.    Israel BE, Tilghman SL, Parker Lemieux K, Payton Stewart F. Phytochemicals: Current strategies for treating breast cancer. Oncol. Lett. 2018;15(5):7471-8. doi: 10.3892/ol.2018.8304
42.    Swarnalatha Y. Isolation of flavonoids and their anticancer activity from Sphaeranthus amaranthoides in A549 cell line. Res. J. Pharm. Technol. 2015;8(4):462-67. doi: 10.5958/0974-360X.2015.00077.3
43.    Hadi SM, Bhat SH, Azmi AS, Hanif S, Shamim U, Ullah MF. Oxidative breakage of cellular DNA by plant polyphenols: a putative mechanism for anticancer properties. Semin. Cancer Biol.  2007;17(5):370-6. doi: 10.1016/j.semcancer.2007.04.002
44.    Ahmed SI, Hayat MQ, Tahir M, Mansoor Q, Ismail M, Keck K, Bates RB. Pharmacologically active flavonoids from the anticancer, antioxidant and antimicrobial extracts of Cassia angustifolia Vahl. BMC Complement Altern. Med. 2016;16(1):1-9. doi:10.1186/s12906-016-1443-z
45.    Grigalius I, Petrikaite V. Relationship between antioxidant and anticancer activity of trihydroxyflavones. Molecules. 2017;22(12):2169. doi: 10.3390/molecules22122169
46.    Baby B, Antony P, Vijayan R. Antioxidant and anticancer properties of berries. Crit. Rev. Food Sci. Nutr. 2018;58(15):2491-507. doi:10.1080/10408398.2017.1329198

Recomonded Articles:

Research Journal of Pharmacy and Technology (RJPT) is an international, peer-reviewed, multidisciplinary journal.... Read more >>>

RNI: CHHENG00387/33/1/2008-TC                     
DOI: 10.5958/0974-360X 

1.3
2021CiteScore
 
56th percentile
Powered by  Scopus


SCImago Journal & Country Rank


Recent Articles




Tags


Not Available