DIFFERENTIAL EFFECT OF PURIFIED QUERCETIN AND ITS DERIVATIVES FROM IN VITRO CELL SUSPENSION CULTURES OF CAESALPINIA PULCHERRIMA SW. AGAINST SELECTED CANCER CELL LINES AND ITS MODE OF ACTION

  • JM Aswathy Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala
  • Greeshma Murukan Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala
  • Bosco Lawarence Department of Botany and Biotechnology, Government Arts College, Thiruvananthapuram, India *e-mail address: harimurukan@gmail.com
  • K Murugan Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala

Abstract

Tribal people use the floral extract of Caesalpinia pulcherrima to cure liver, stomach and skin prone disorders in traditional Indian medicine. This study aimed to evaluate the effect of purified quercetin and its derivatives from in vitro cell suspension cultures of C. pulcherrima Sw. against SW 480, HeLa, MCF-7 and MCF 10A cell lines and its mode of action. Standard protocol was developed for callus induction using leaf explants. Cytotoxic effect was evaluated against SW 480, HeLa, MCF-7 and MCF 10A cells by MTT assay. Apoptosis was evaluated via Hoechst analysis,  flow cytometry, mitochondrial membrane potential and caspase 3 and 9 expression. 2, 4-D (2.5 mg/l), BAP (2.5 mg/l) + kin (1 mg/ml) was effective for remarkable callus induction. Further, cell suspension culture was established.  Effect of elicitors on cell suspension culture was also carried. Sucrose, ABA and salicylic acid (SA) at different concentrations influenced cell biomass and quercetin synthesis. Cells cultured on the medium fortified with 45 g/L sucrose without ABA/SA showed the highest quercetin content (16.5 mg/g). Quercetin was purified, fractionated by HPLC-DAD and was further analyzed by NMR revealed a major fraction of quercetin (3, 5, 7, 3’, 4’-pentahydroxyflavon). Insignificant cytotoxicity was noticed in SW 480, HeLa, MCF 10A when compared to MCF-7 cell lines exposed to different concentrations of purified quercetin for 24- 48 h. Similarly, the apoptosis by nuclei staining using Hoechst 33258 revealed a concentration dependent effect on MCF 7 cells only. This was further substantiated by caspase-9 and 3 induction and mitochondrial depolarization as revealed by flow cytometry. Overall, the results showed that quercetin and its derivatives induced effective apoptosis on MCF-7 cells. Quercetin isolated from the in vitro cell suspension culture of C. pulcherrima showed significant cytotoxicity and apoptotic activity towards MCF-7 cell lines as compared to other cell lines.


KeywordsCaesalpinia pulcherrima; quercetins; suspension culture; cytotoxicity; apoptotic.

Downloads

Download data is not yet available.

Author Biographies

JM Aswathy, Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala

Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala

Greeshma Murukan, Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala

Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala

Bosco Lawarence, Department of Botany and Biotechnology, Government Arts College, Thiruvananthapuram, India *e-mail address: harimurukan@gmail.com

Department of Botany and Biotechnology, Government Arts College, Thiruvananthapuram, India

*e-mail address: harimurukan@gmail.com

K Murugan, Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala

Plant biochemistry and molecular biology laboratory, Department of Botany, University College, Thiruvananthapuram 695034, Kerala

References

1. Balasuriya BWN, Rupasinghe HPV, Plant quercetins as angiotensin converting enzyme inhibitors in regulation of hypertension, Funct. Foods Health Dis, 2011; 5:172–188.
2. Batra P, Sharma AK, Anti-cancer potential of quercetins: recent trends and future perspectives, 3 Biotech, 2013; 3(6):439–459.
3. Bode AM, Dong Z, Post-translational modification of p53 in tumorigenesis, Nat. Rev. Cancer. 2004; 4:793–805.
4. Ho JS, Ma W, Mao DY, Benchimol S, p53-dependent transcriptional repression of c-myc is required for G1 cell cycle arrest, Mol. Cell Bio, 2005; 25:7423–7431.
5. Amin AR, Kucuk O, Khuri FR, Shin DM, Perspectives for cancer prevention with dietary compounds, J. Clin. Oncol, 2009; 27(16):2712–2725.
6. Roy AM, Baliga MS, Katiyar SK, Epigallocatechin-3-gallate induces apoptosis in estrogen receptor-negative human breast carcinoma cells via modulation in protein expression of p53 and Bax and caspase-3 activation, Mol. Cancer Ther, 2005; 4:81–90.
7. Patil SJ, Salunkhe VR, Alai MH, Estimation of quercetin in ayurvedic proprietary medicine by UV- spectrophotometry, Int. J. Pharm. Pharm. Sci, 2012; 4(3):645-647.
8. Le Son H, Le Quoc Linh N, Minh T V, Isolation and in vitro anticancer activity of flavonoids from Caesalpinia sappan Linn, J. Sci, 2015; 3(3):24-30.
9. Anju V, Zachariah S M, Phytochemical screening, isolation, antibacterial and anticancer activity studies of Caesalpinia pulcherrima Linn. leaves by HPTLC analysis, Int. J. Pharm. Bio Sci, 2017; 8(2):12-29.
10. Shivaprakash P, Balaji KS, Lakshmi GM, Chandrashekara KT, Jayarama S, Methanol extract of Caesalpinia bonducella induces apoptosis via up-regulation of Bax and activation of PARP in Ehrlich Ascites tumor cells, Med. Aromat. Plants, 2016; 5:273.
11. Pawar C R, Mutha R E, Landge A D, Jadhav R B, Surana S J, Antioxidant and cytotoxic activities of Caesalpinia pulcherrima wood, Ind. J. Biochem. Biophy, 2009; 46:198-200.
12. Zanin J L B, Massoni M, dos Santos M H, de Freitas G, Niero E L O, Schefer R R, Lago J H G, Iontae M, Soares M G. Caesalpinioflavone, a New Cytotoxic Biflavonoid Isolated from Caesalpinia pluviosa var. peltophoroides. J. Braz. Chem. Soc. 2015; 26(4):804-809.
13. Orangi M, Pasdaran A, Shanehbandi D, Kazemi T, Yousefi B, Hosseini B, Baradaran B, Cytotoxic and apoptotic activities of methanolic subfractions of Scrophularia oxysepala against human breast cancer cell line, Evid. Based Complement Alternat. Med, 2016:8540640.
14. Looi CY, Arya A, Cheah FK, Muharram B, Leong KH, Mohamad K, Wong WF, Rai N, Mustafa MR, Induction of apoptosis in human breast cancer cells via caspase pathway by vernodalin isolated from Centratherum anthelminticum (L.) Seeds PLoS One, 2013; 8(2):e56643.
15. Hosseini BA, Pasdaran A, Kazemi T, Shanehbandi D, Karami H, Orangi M, Baradaran B, Dichloromethane fractions of Scrophularia oxysepala extract induce apoptosis in MCF-7 human breast cancer cells, Bosn. J. Basic Med. Sci, 2015; 15(1):26-32.
16. Shoja MH, Reddy ND, Nayak PG, Srinivasan K, Rao CM, Glycosmis pentaphylla (Retz.) DC arrests cell cycle and induces apoptosis via caspase-3 activation in breast cancer cells, J. Ethnopharmaco, 2015; 168(20):50-60.
17. Vijayalakshmi A , Masilamani K , Nagarajan E and Ravichandiran V, In vitro antioxidant and anticancer activity of quercetins from Cassia tora Linn. leaves against human breast carcinoma cell lines, Der Pharma Chemica, 2015; 7(9):122-129.
18. Sak K, Cytotoxicity of dietary quercetins on different human cancer types, Pharmacogn. Rev, 2014; 8(6): 122-146.
19. Priya K, Neha B, Robinka K, Quercetins and their therapeutic potential as anticancer agents: biosynthesis, metabolism and regulation, World J. Pharm. Pharm. Sci, 2014; 3(6):2188-2216.
20. Rafik S, Mahesh P, Ashwini D, Sayyed I, Evaluation of anticancer, antioxidant and possible anti-inflammatory properties of selected medicinal plants used in Indian traditional medication, J. Tradit. Complement. Med, 2014; 4(4):253-257.
21. Hoang LS, Nquyem Le QL, Tran VM, Isolation and in vitro anticancer activity of quercetins from Ceasalpinia sappan L, J. Sci, 2015; 3(3):24-30.
22. Zainab YM, Khulood WA, Subhi JH, Preliminary study for the anticancer activity of quercetins extracted from wild Lycium barbarum leaves, Int. J. Res. Stud. Biosci, 2015; 3(12):88-94.
23. Chandrappa CP, Govindappa M, Anil Kumar NV, Channabasava R, Chandrasekar N, Umashankar T, Identification and separation of quercetin from ethanol extract of Carmona retusa by thin layer chromatography and high performance liquid chromatography with diode array detection, World J. Pharm. Pharm. Sci, 2014; 3 (6):2020-2029.
24. Nancy ER, Chi-TangHo, ShimingLi, Efficacious anti-cancer property of quercetins from citrus peels, Food Sci. Human Well, 2014; 3:104-109.
25. Shikha S, Ranganatha RS, Mahesh H, Mayilaadumveettil N, Satish KT, Mrinal S, Quercetin, a Natural Quercetin Interacts with DNA, Arrests Cell Cycle and Causes Tumor Regression by Activating Mitochondrial Pathway of Apoptosis, Scientific reports, 2016.
26. Joby J, Sudheesh S, Sumesh KT, Sony J, Jayadevi V, A comparative evaluation of anticancer activities of quercetins isolated from Mimosa pudica, Aloe vera and Phyllanthus niruri against human breast carcinoma cell line (MCF-7) using MTT assay, Int. J. Pharm. Pharm. Sci, 2014; 6(2):319-322.
Statistics
45 Views | 0 Downloads
How to Cite
Aswathy, J., Murukan, G., Lawarence, B., & Murugan, K. (2018). DIFFERENTIAL EFFECT OF PURIFIED QUERCETIN AND ITS DERIVATIVES FROM IN VITRO CELL SUSPENSION CULTURES OF CAESALPINIA PULCHERRIMA SW. AGAINST SELECTED CANCER CELL LINES AND ITS MODE OF ACTION. Journal of Drug Delivery and Therapeutics, 8(5-s), 196-208. https://doi.org/10.22270/jddt.v8i5-s.1950
Section
Research