Login Register Cart 0
Generic placeholder image

The Natural Products Journal

Editor-in-Chief

ISSN (Print): 2210-3155
ISSN (Online): 2210-3163

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Apoptosis Inducing Effects of Thymus Linearis Methanolic Extract in HCT-116 Cells and LC-MS Chemical Profiling of Its Active Constituents

Author(s): Rohina Bashir, Ovais Zargar, Qazi Parvaiz and Rabia Hamid*

Volume 12, Issue 4, 2022

Published on: 28 January, 2021

Article ID: e160921190858 Pages: 12

DOI: 10.2174/2210315511999210128202816

Price: $65

Open Access Journals Promotions 2
Abstract

Background: Cancer is one of the major problems at present, to which vast research is being dedicated to find an effective remedy. Medicinal plants are endowed with numerous molecules that could be effective in multiple diseases including cancer. Thymus linearis, being rich in phenols, terpenoid, and flavonoids have the potential to provide anti-cancer entities.

Methods: The extracts of Thymus linearis were investigated for in vitro anticancer activity using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay on a panel of cancer cell lines. The cellular and nuclear morphology was studied using microscopic techniques. Agarose gel electrophoresis was used for DNA fragmentation analysis. Protein expression was determined by western blotting. LC-MS was used for phytochemical identification.

Results: Among all the extracts, Thymus linearis methanolic (TLM) extract was found to exhibit antiproliferative activity on cell lines to varying degrees. TLM was found to be most potent against HCT-116 with an IC50 of 158μg/ml after 48hrs treatment while being nontoxic to HEK-293 and FR-2 cells under similar concentrations. TLM decreased clonogenic potential of HCT-116 cells. It induced cell shrinkage, membrane blebbing and nuclear fragmentation characteristic of apoptotic in a dose dependent manner in HCT-116 cells. Prominent internucleosomal DNA cleavage was observed in HCT-116 cells after 48hrs TLM treatment. Western blot analysis revealed the up regulation of expression of Bax, caspases 9 and caspases 3 and downregulation of Bcl-2 proteins. The LC-MS data revealed the presence of Salvianolic acid H, Synparvolide C, Thymuside A and Jasmonic acid; 12-Hydroxy, O-β-D-glucopyranoside and polyphenolic flavonoids to which antiproliferative activity can be attributed.

Conclusion: The results suggest that Thymus linearis methanolic extract could be valuable source of anti-cancer agents.

Keywords: Thymus linearis, colon cancer, HEK-293, MTT, LC-MS, Bcl-2, apoptosis.

Graphical Abstract

[1]
Gerlinger, M.; Rowan, A.J.; Horswell, S.; Math, M.; Larkin, J.; Endesfelder, D.; Gronroos, E.; Martinez, P.; Matthews, N.; Stewart, A.; Tarpey, P.; Varela, I.; Phillimore, B.; Begum, S.; McDonald, N.Q.; Butler, A.; Jones, D.; Raine, K.; Latimer, C.; Santos, C.R.; Nohadani, M.; Eklund, A.C.; Spencer-Dene, B.; Clark, G.; Pickering, L.; Stamp, G.; Gore, M.; Szallasi, Z.; Downward, J.; Futreal, P.A.; Swanton, C. Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N. Engl. J. Med., 2012, 366(10), 883-892.
[http://dx.doi.org/10.1056/NEJMoa1113205] [PMID: 22397650]
[2]
Siegel, R.L.; Miller, K.D.; Jemal, A. Cancer statistics, 2016. CA Cancer J. Clin., 2016, 66(1), 7-30.
[http://dx.doi.org/10.3322/caac.21332] [PMID: 26742998]
[3]
Cheng, Y-T.; Yang, C-C.; Shyur, L-F. Phytomedicine-Modulating oxidative stress and the tumor microenvironment for cancer therapy. Pharmacol. Res., 2016, 114, 128-143.
[http://dx.doi.org/10.1016/j.phrs.2016.10.022] [PMID: 27794498]
[4]
Xie, S.; Zhou, J. Harnessing plant biodiversity for the discovery of novel anticancer drugs targeting microtubules. Front. Plant Sci., 2017, 8, 720.
[http://dx.doi.org/10.3389/fpls.2017.00720] [PMID: 28523014]
[5]
Pfeffer, C.M.; Singh, A.T.K. Apoptosis: A target for anticancer therapy. Int. J. Mol. Sci., 2018, 19(2), 448.
[http://dx.doi.org/10.3390/ijms19020448] [PMID: 29393886]
[6]
Pedersen, J.A. Distribution and taxonomic implications of some phenolics in the family Lamiaceae determined by ESR spectroscopy. Biochem. Syst. Ecol., 2000, 28(3), 229-253.
[http://dx.doi.org/10.1016/S0305-1978(99)00058-7]
[7]
Banno, N.; Akihisa, T.; Tokuda, H.; Yasukawa, K.; Higashihara, H.; Ukiya, M.; Watanabe, K.; Kimura, Y.; Hasegawa, J.; Nishino, H. Triterpene acids from the leaves of Perilla frutescens and their anti-inflammatory and antitumor-promoting effects. Biosci. Biotechnol. Biochem., 2004, 68(1), 85-90.
[http://dx.doi.org/10.1271/bbb.68.85] [PMID: 14745168]
[8]
Rana, S.S.; Angiras, N.N.; Sharma, G.D. Effect of herbicides and interculture on nutrient uptake by puddle seeded rice and associated weeds. Indian J. Weed Sci., 2000, 32(1/2), 70-73.
[9]
Rana, P.K. Uses of local plant biodiversity among the tribal communities of Pangi Valley of district Chamba in cold desert Himalaya, India. Sci. World J., 2014, 2014, 753289.
[http://dx.doi.org/10.1155/2014/753289]
[10]
Hussain, A.I. Chemical composition and bioactivity studies of the essential oils from two Thymus species from the Pakistani flora. Lebensm. Wiss. Technol., 2013, 50(1), 185-192.
[http://dx.doi.org/10.1016/j.lwt.2012.06.003]
[11]
Alamgeer, ; Akhtar, M.S.; Jabeen, Q.; Khan, H.U.; Maheen, S.; Haroon-Ur-Rash, ; Karim, S.; Rasool, S.; Malik, M.N.; Khan, K.; Mushtaq, M.N.; Latif, F.; Tabassum, N.; Khan, A.Q.; Ahsan, H.; Khan, W. Pharmacological evaluation of antihypertensive effect of aerial parts of Thymus linearis benth. Acta Pol. Pharm., 2014, 71(4), 677-682.
[PMID: 25272894]
[12]
Alamgeer, ; Auger, C.; Chabert, P.; Lugnier, C.; Mushtaq, M.N.; Schini-Kerth, V.B. Mechanisms underlying vasorelaxation induced in the porcine coronary arteries by Thymus linearis, Benth. J. Ethnopharmacol., 2018, 225, 211-219.
[http://dx.doi.org/10.1016/j.jep.2018.07.010] [PMID: 30009977]
[13]
Monks, A.; Scudiero, D.; Skehan, P.; Shoemaker, R.; Paull, K.; Vistica, D.; Hose, C.; Langley, J.; Cronise, P.; Vaigro, W.A. Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines. J. Nat. Cancer Inst., 1991, 83(11), 757-766.
[http://dx.doi.org/10.1093/jnci/83.11.757] [PMID: 2041050]
[14]
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.
[http://dx.doi.org/10.1016/0022-1759(83)90303-4] [PMID: 6606682]
[15]
Gupta, D.; Crosby, M.E.; Almasan, A.; Macklis, R.M. Regulation of CD20 expression by radiation-induced changes in intracellular redox status. Free Rad. Biol. Med., 2008, 44(4), 614-623.
[http://dx.doi.org/10.1016/j.freeradbiomed.2007.10.048] [PMID: 18060882]
[16]
Lambu, M.R.; Kumar, S.; Yousuf, S.K.; Sharma, D.K.; Hussain, A.; Kumar, A.; Malik, F.; Mukherjee, D. Medicinal chemistry of dihydropyran-based medium ring macrolides related to aspergillides: Selective inhibition of PI3Kα. J. Med. Chem., 2013, 56(15), 6122-6135.
[http://dx.doi.org/10.1021/jm400515c] [PMID: 23845251]
[17]
Rahbar Saadat, Y.; Saeidi, N.; Zununi Vahed, S.; Barzegari, A.; Barar, J. An update to DNA ladder assay for apoptosis detection. Bioimpacts, 2015, 5(1), 25-28.
[http://dx.doi.org/10.15171/bi.2015.01] [PMID: 25901294]
[18]
Ghasemi, A.; Khanzadeh, T.; Zadi Heydarabad, M.; Khorrami, A.; Jahanban Esfahlan, A.; Ghavipanjeh, S.; Gholipour Belverdi, M.; Darvishani Fikouhi, S.; Darbin, A.; Najafpour, M.; Azimi, A. Evaluation of BAX and BCL-2 gene expression and apoptosis induction in acute lymphoblastic leukemia cell Line CCRFCEM after high- dose prednisolone treatment. Asian Pac. J. Cancer Prev., 2018, 19(8), 2319-2323.
[PMID: 30141309]
[19]
Li, J.; Yuan, J. Caspases in apoptosis and beyond. Oncogene, 2008, 27(48), 6194-6206.
[http://dx.doi.org/10.1038/onc.2008.297] [PMID: 18931687]
[20]
Liu, X.; Yang, Y.; Zhang, X.; Xu, S.; He, S.; Huang, W.; Roberts, M.S. Compound Astragalus and Salvia miltiorrhiza extract inhibits cell invasion by modulating transforming growth factor-β/Smad in HepG2 cell. J. Gastroenterol. Hepatol., 2010, 25(2), 420-426.
[http://dx.doi.org/10.1111/j.1440-1746.2009.05981.x] [PMID: 19793165]
[21]
Tsai, M-K.; Lin, Y-L.; Huang, Y-T. Effects of salvianolic acids on oxidative stress and hepatic fibrosis in rats. Toxicol. Appl. Pharmacol., 2010, 242(2), 155-164.
[http://dx.doi.org/10.1016/j.taap.2009.10.002] [PMID: 19822164]
[22]
Ho, J.H-C.; Hong, C-Y. Salvianolic acids: small compounds with multiple mechanisms for cardiovascular protection. J. Biomed. Sci., 2011, 18(1), 30.
[http://dx.doi.org/10.1186/1423-0127-18-30] [PMID: 21569331]
[23]
Wang, T.-y.; Li, Q.; Bi, K.-s. Bi, Bioactive flavonoids in medicinal plants: Structure, activity and biological fate. Asian j. of Pharmaceuti Sci, 2017.
[24]
Krych, J.; Gebicka, L. Catalase is inhibited by flavonoids. Int. J. Biol. Macromol., 2013, 58, 148-153.
[http://dx.doi.org/10.1016/j.ijbiomac.2013.03.070] [PMID: 23567286]
[25]
Grimmer, H.R.; Parbhoo, V.; McGrath, R.M. Antimutagenicity of polyphenol-rich fractions from sorghum bicolor grain. J. Sci. Food Agric., 1992, 59(2), 251-256.
[http://dx.doi.org/10.1002/jsfa.2740590217]
[26]
Patridge, E.; Gareiss, P.; Kinch, M.S.; Hoyer, D. An analysis of FDA-approved drugs: Natural products and their derivatives. Drug Discov. Today, 2016, 21(2), 204-207.
[http://dx.doi.org/10.1016/j.drudis.2015.01.009] [PMID: 25617672]
[27]
Miranda-Vilela, A.L.; Portilho, F.A.; de Araujo, V.G.; Estevanato, L.L.; Mezzomo, B.P.; Santos, Mde.F.; Lacava, Z.G. The protective effects of nutritional antioxidant therapy on Ehrlich solid tumor-bearing mice depend on the type of antioxidant therapy chosen: Histology, genotoxicity and hematology evaluations. J. Nutr. Biochem., 2011, 22(11), 1091-1098.
[http://dx.doi.org/10.1016/j.jnutbio.2010.09.009] [PMID: 21273055]
[28]
Franken, N.A.P.; Rodermond, H.M.; Stap, J.; Haveman, J.; Bree, C. Clonogenic assay of cells in vitro. Nat. Protoc., 2006, 1(5), 2315-2319.
[http://dx.doi.org/10.1038/nprot.2006.339] [PMID: 17406473]
[29]
Zaker, A.; Asili, J.; Abrishamchi, P.; Tayarani-Najaran, Z.; Mousavi, S.H. Cytotoxic and apoptotic effects of root extract and tanshinones isolated from Perovskiaabrotanoides Kar. Iran. J. Basic Med. Sci., 2017, 20(12), 1377-1384.
[PMID: 29238474]
[30]
Pieme, C.A.; Kumar, S.G.; Dongmo, M.S.; Moukette, B.M.; Boyoum, F.F.; Ngogang, J.Y.; Saxena, A.K. Antiproliferative activity and induction of apoptosis by Annona muricata (Annonaceae) extract on human cancer cells. BMC Complement. Alter. Med., 2014, 14(1), 516.
[http://dx.doi.org/10.1186/1472-6882-14-516] [PMID: 25539720]
[31]
Khazaei, S. Cytotoxicity and proapoptotic effects of Allium atroviolaceum flower extract by modulating cell cycle arrest and caspase-dependent and p53-independent pathway in breast cancer cell lines. Evid. Based Complement. Alter. Med., 2017, 2017, 1-16.
[http://dx.doi.org/10.1155/2017/1468957]
[32]
Elkady, A.I.; Hussein, R.A.E.H.; Abu Zinadah, O.A. Differential control of growth, apoptotic activity and gene expression in human colon cancer cells by extracts derived from medicinal herbs, Rhazya stricta and Zingiber officinale and their combination. World J. Gastroenterol., 2014, 20(41), 15275-15288.
[http://dx.doi.org/10.3748/wjg.v20.i41.15275] [PMID: 25386076]
[33]
Miah, M. Methanolic Bark Extract of Abroma augusta (L.) Induces Apoptosis in EAC Cells through Altered Expression of Apoptosis Regulatory Genes. Evid. Based. Complement. Alter. Med., 2020, 2020, 9145626.
[34]
Bayala, B.; Bassole, I.H.; Scifo, R.; Gnoula, C.; Morel, L.; Lobaccaro, J.M.; Simpore, J. Anticancer activity of essential oils and their chemical components - A review. Am. J. Cancer Res., 2014, 4(6), 591-607.
[PMID: 25520854]
[35]
Qin, T.; Rasul, A.; Sarfraz, A.; Sarfraz, I.; Hussain, G.; Anwar, H.; Riaz, A.; Liu, S.; Wei, W.; Li, J.; Li, X. Salvianolic acid A & B: Potential cytotoxic polyphenols in battle against cancer via targeting multiple signaling pathways. Int. J. Biol. Sci., 2019, 15(10), 2256-2264.
[http://dx.doi.org/10.7150/ijbs.37467] [PMID: 31592132]
[36]
Wang, T.Y.; Li, Q.; Bi, K.-s. Bioactive flavonoids in medicinal plants: Structure, activity and biological fate. Asian. J. Pharm. Sci., 2018, 13(1), 12-23.

Rights & Permissions Print Cite
Article Metrics
27
1
Wayfinder Image
Open Access Journals Promotions
© 2024 Bentham Science Publishers | Privacy Policy