Chemotherapy being a significant treatment methodology utilized for the control of cutting edge phases of malignancies and as a prophylactic against conceivable metastasis, shows extreme poisonousness on typical tissues.⁹

The main case-controlled examination on bosom malignant growth the study of disease transmission was finished by Janet Lane-Claypon, who distributed a relative report in 1926 of 500 bosom malignancy cases and 500 controls of a similar foundation and way of life for the British Ministry of Health.^.¹⁰

During the 1980s and 1990s, a great many ladies who had effectively finished standard treatment at that point requested and got high-portion bone marrow transplants, figuring this would prompt better long haul endurance. In any case, it demonstrated totally incapable, and 15–20% of ladies kicked the bucket in view of the merciless treatment.¹¹

The 1995 reports from the Nurses' Health Study and the 2002 finishes of the Women's Health Initiative preliminary decisively demonstrated that hormone substitution treatment altogether expanded the frequency of bosom cancer.

Bosom malignant growth is the most widely recognized female disease overall speaking to about a quarter (23%) of all tumors in ladies^12,13. The worldwide weight of bosom disease is relied upon to cross 2 million continuously 2030, with developing extents from creating nations¹⁴. In spite of the fact that age-institutionalized frequency rates in India are lower than in the United Kingdom (UK) (25.8 versus 95 for every 100, 000), death rates are about as high (12.7 versus 17.1 per 100, 000, separately) as those of the UK. Bosom malignant growth occurrence rates inside India show a 3–4-crease variety the nation over, with the most noteworthy rates saw in the Northeast and in significant metropolitan urban areas, for example, Mumbai and New Delhi¹⁵. Explanations behind this variety remember contrasts for segment (e.g., training), conceptive (e.g., age from the outset kid and number of kids), anthropometric (e.g., adiposity) and way of life factors (e.g., tobacco smoking and liquor use)¹⁶.

Restorative plants speak to a financially significant and solid part of biodiversity segments. It is important to do a full stock of restorative plants in any nation vegetation so as to secure and guarantee the progression of utilization and along these lines ensure jeopardized therapeutic species in nature. The utilization of restorative plants is viewed as increasingly basic in conventional medication and pharmaceutical device beneficial around the world. Numerous people group bolster characteristic items gathered from environments for therapeutic purposes just as for nourishment purposes.¹⁷

Plants have been utilized for treating different ailments of people and creatures since days of yore. They keep up the wellbeing and essentialness of people, and furthermore fix illnesses, including malignancy without causing poisonousness. Over half of every cutting edge sedate in clinical use are of normal items, huge numbers of which can control malignant growth cells.¹⁸

The world wellbeing association (WHO) has assessed that 80% of the world's occupant depended on customary medication for their essential human services needs and the greater part of these treatments included the utilization of plant extricate or their dynamic mixes (Bruneton, 1995). Justicia adhatoda (L.) Nees (family Acanthaceae) is a bush across the board all through the tropical areas of Southeast Asia.¹⁹

INTRODUCTION OF THYMOL:

Trachyspermum ammi is a local of Egypt and is developed in Iraq, Iran, Afghanistan, Pakistan, and India. In India, it is developed in Madhya Pradesh, Uttar Pradesh, Gujarat, Rajasthan, Maharashtra, Bihar and West Bengal²⁰. Trachyspermum ammi L. having a place with family Apiaceae is a profoundly esteemed restoratively significant seed zest. The roots are diuretic in nature and the seeds have superb Spanish fly properties. The seeds contain 2–4.4% earthy colored shaded oil known as ajwain oil. The fundamental part of this oil is thymol, which is utilized in the treatment of gastro-intestinal diseases, absence of craving and bronchial issues. The oil displays fungicidal, antimicrobial and against aggregatory impacts on humans. Ajwain is a customary potential herb and is broadly utilized for restoring different infections in people and creatures. The natural product has energizer, antispasmodic and carminative properties. It is a significant therapeutic specialist for fart, atonic dyspepsia and diarrhea²¹. The seed of ajwain is unpleasant, sharp and it goes about as anthelmintic, carminative, diuretic, and stomachic. It additionally fixes stomach tumors, stomach torments and piles. Seeds contain a basic oil.

In Ayurvedic arrangement of medication it is fundamentally utilized in the treatment of respiratory, gastro-intestinal clutters and incendiary conditions.²²

Fig 1.2 Ajwain (Trachyspermum ammi)

Thymol, a characteristic monoterpene phenol subordinate of cymene (C10H14O), is synthetically known as 2-isopropyl-5-methylphenol. Thymol, a fundamental oil is biosynthesized through the transformation of gamma-terpinene to p-cymene²³. It is affirmed by EU and FDA as a protected nourishment added substance. The basic herbal wellsprings of thymol are Euphrasia rostkoviana²⁴, Monarda didyma, M. fistulosa, Nigella sativa, Trachyspermum ammi, Origanum compactum, O. dictamnus, O. onites, O. vulgare, Thymus glandulosus, T. hyemalis, T. vulgaris, and T. zygis.

Thymol and its subsidiaries are broadly utilized in clinical practices, beauty care products, nourishment, and farming industry²⁵. In pharmacology, a medication is "a compound substance utilized in the treatment, fix, anticipation, or analysis of malady or used to in any case upgrade physical or mental prosperity. Thymol has different organic exercises, including cancer prevention agent, mitigating, antimicrobial, anti diabetic, anti obesity, neuro pharmacological, anti hyperglycemic, and hypolipidemic impacts²⁶.

Thymol shows unconstrained contractile action (SCA) of smooth muscle strips (SCA) from the stomach and vena portae of guinea pigs.²⁷

From the early reports thymol and its subsidiaries are obvious for its anticancer, immune modulatory activities. Thymol and its subsidiaries have been accounted for to exert their anticancer action through an alternate instrument of activity including smothering cell development, inciting apoptosis delivering intracellular receptive oxygen species, depolarizing mitochondrial film potential, and initiating the proapoptotic mitochondrial proteins Bax, association with cysteine aspartases (caspases) or poly ADP ribose polymerase in various test model.²⁸

Anticancer action of orchestrated mixes was controlled by MTT test and NR measure strategy utilizing bosom malignant growth cell lines (MCF-7 and MDA MB 231)²⁹.

Chemistry of thymol:

Thymol (also known as 2-isopropyl-5-methylphenol, IPMP) is a natural monoterpenoid phenol derivative of cymene, C₁₀H₁₄O, isomeric with carvacrol, found in oil of thyme, and extracted from Thymus vulgaris (common thyme), Ajwain and various other kinds of plants as a white crystalline substance of a pleasant aromatic odor and strong antiseptic properties. Thymol is only slightly soluble in water at neutral pH, but it is extremely soluble in alcohols and other organic solvents. It is also soluble in strongly alkaline aqueous solutions due to deprotonation of the phenol.

Fig 1.3 Thymol

Anti Cancer mechanism of thymol:

Plants are the wellsprings of various malignant growth chemotherapeutic specialists. The greater part of malignant growth chemotherapeutic medications act through the double pro‐oxidant and cancer prevention agent impacts, the mitochondrial brokenness pathway of disease cell executing through which produces receptive oxygen species (ROS), the provocative instrument, and cell cycle guideline³⁰. Mastelić et al. conveyed an examination, in which thymol (100–600μg/mL) was clear to show critical DPPH rummaging limit. Right now, creators likewise announced an antioxidant‐dependent anti proliferative impact at 0.1–100mM of thymol in human cervical carcinoma cells (HeLa). Thymol (75–1, 200 μM) can create a stable phenoxy radical middle of the road, which produces ROS and quinone oxide subsidiaries in melanoma (B16‐F10) cells³¹. In an ongoing report, thymol secured the medication safe H1299 cells against hydrogen peroxide (H2O2)‐induced cytotoxicity, and layer and DNA harm. Moreover, thymol ensured human hepato carcinoma (HepG2) cells by rummaging free radicals, in this way diminishing oxidative pressure and mitochondrial harm along these lines hindering cell demise³².

Cancer prevention agents are defensive at a low fixation/portion while pro‐oxidant or cytotoxic at high focus/portion. For a model, ascorbic corrosive is a cell reinforcement at low focus while pro‐oxidant at high fixation³³. Notwithstanding, the pro‐oxidative impacts of a cancer prevention agent may change because of varieties of test frameworks, time of introduction, cell inward obstructions, etc. Thymol (25–150μM) indicated ROS‐induced mitochondrial cell demise and DNA harm, up‐regulated poly [ADP‐ribose] polymerase 1 (PARP‐1), phospho c‐Jun N‐terminal kinase (p‐JNK), cytochrome c and caspase‐3 proteins in HCT‐116 cells, while at 25, 50, and 50μM it caused an apoptotic cell passing through the characteristic pathway alongside ROS enlistment, caspase‐3/9 initiation, mitogen‐activated protein kinase (MAPK) actuation, arrival of cytochrome c and down‐regulation of antiapoptotic Bcl‐2 family proteins in bladder malignancy cells. Thymol (10–400μM) applied cytotoxic, genotoxic, apoptotic, ROS‐generating, and decreased glutathione (GSH)‐reducing impacts in gastric adenocarcinoma (AGS) cells³⁴.

MATERIALS AND METHODS:

Reagent and chemicals:

Thymol was bought from Sigma Aldrich. Stock arrangement of thymol was set up in dimethyl sulfoxide (DMSO) and was weakened to conclusive fixation in the way of life medium. Last focus of DMSO utilized as vehicle never surpassed 0.03% and had no perceptible consequences for MCF-7, SK-MEL-2, HCT-15 and MIAPaCa-2 cells in examination with the untreated control. Tissue culture medium constituents, all synthetic concoctions and solvents were expository evaluation bought from Hi Media (Mumbai, India).

Cell culture:

The human bosom disease cell line MCF-7 cell line was given by National Center to Cell Science (NCCS), Pune and was developed in Eagles Minimum Essential Medium (EMEM) which contained 10% fetal ox-like serum (FBS). All cells were kept up at 37°C, 100% relative moistness, 5% CO2, 95% air and the way of life medium was changed two times every week.³⁵

Cytotoxicity assay:

Cytotoxicity movement was assessed by Sulforhodamine B (SRB) examine as depicted by Vichai et al. Exploratory medications were solubilized in suitable dissolvable to get ready supply of 10-2 focus. At the hour of test four 10-crease sequential weakenings were made utilizing complete medium. Aliquots of 10µL of these distinctive medication weakenings were added to the proper smaller scale titer wells previously containing 90 µL of medium, bringing about the necessary last medication fixations.

After expansion of aggravates; the plates were hatched at standard conditions for 48 hours and measure was ended by the expansion of cold TCA. Cells were fixed in-situ by the delicate expansion of 50 µL of cold 30% (w/v) TCA (last fixation, 10% TCA) and hatched for an hour at 4°C. The supernatant was disposed of; the plates were washed multiple times with faucet water and air dried. Sulforhodamine B (SRB) arrangement (50µL) at 0.4% (w/v) in 1% acidic corrosive was added to each of the wells, and plates were brooded for 20 minutes at room temperature. Subsequent to recoloring, unbound color was recouped and the lingering color was evacuated by washing multiple times with 1% acidic corrosive. The plates were air dried. Bound stain was along these lines eluted with 10 mM trizma base, furthermore, the absorbance was perused on an Elisa plate peruse at a frequency of 540 nm with 690 nm reference frequency.

Percent development was determined on a plate-by-plate reason for test wells comparative with control wells. Percent development was communicated as the proportion of normal absorbance of the test well to the normal absorbance of the control wells × 100. Utilizing the six absorbance estimations [time zero (Tz), control development (C), and test development within the sight of medicate at the four focus levels (Ti)], the rate development was determined at every one of the medication focus levels. The portion reaction parameters were determined for each test article. Development hindrance of half (GI50) was determined from [(Ti-Tz)/(C-Tz)] × 100=50, which is the medication focus bringing about a half decrease in the net protein increment (as estimated by SRB recoloring) in control cells during the medication brooding. The medication focus bringing about complete development restraint (TGI) was determined from Ti=Tz. The LC50 (grouping of medication bringing about a half decrease in the deliberate protein toward the finish of the medicate treatment when contrasted with that toward the start) showing a net loss of cells following treatment is determined from [(Ti-Tz)/Tz] × 100=- 50. Qualities were determined for every one of these three parameters if the level of action was reached; be that as it may, if the impact was not reached or was surpassed, the qualities for that parameter were communicated as more noteworthy or not exactly the most extreme or least fixation tried.

DISCUSSION:

Malignant growth is an exceptionally unpredictable ailment and the event and advancement of tumor cells are firmly identified with strange intracellular sign transduction framework.^{36, 37}These days, one of the fundamental strategies for current malignant growth treatment is chemotherapy. In any case, most chemotherapeutic operators have diverse considerable short-and long haul reactions. Hence, as of late, significant research has been centered around herbs and plants which have been considered for being nontoxic and for the avoidance and treatment of specific sorts of disease.

In the current examination, the concentrate of T. ammi could diminish cell reasonability of MCF-7 bosom disease cell line. TAE (Trachyspermum ammi extract)- incited cell lethality is performed through the acceptance of apoptosis and hindrance of cell multiplication.

The root of malignancy includes the misrepresented cell expansion, just as the concealment of apoptotic forms. Apoptosis is a significant kind of cell passing because of cytotoxic applicant in malignant circumstance. It has been reported that numerous normal mixes with anticancer property could instigate apoptosis of tumor cells.³⁸What's more, various logical reports have exhibited that the acceptance of apoptotic passing in disease treatment is strikingly connected with initiation of caspase 3.³⁹ Since caspase 3 has a focal job in apoptosis and should be the last agent of apoptosis pathway we assessed the impact of TAE on initiated caspase 3 level in MCF-7 cells.

Notwithstanding receptor-intervened apoptosis, there is another pathway actuated by cytotoxic mixes. It happens by adjustment in mitochondrial penetrability and ensuing cytochrome c discharge and arrangement of the apoptosome and initiation of caspase 9 and afterward caspase 3 bringing about downstream occasions engaged with cell demise. It has been uncovered that the arrival of cytochrome c is managed by Bcl2 family proteins. Antiapoptotic Bcl2 relatives exist in the external mitochondrial film and forestall cytochrome c discharge, while proapoptotic individuals are translocated to the mitochondria to instigate apoptosis either by framing pores in mitochondria straightforwardly or by irritating the antiapoptotic proteins.⁴⁰

Cyclin D1, a subunit of CDK4 and CDK6, is one of the major biochemical switches in cell cycle.⁴¹ Upregulation of this utilitarian protein is likewise seen in a few malignancies, including bosom, prostate, neck, and head diseases^{42, 43, 44}. It has been reported that a few phytochemicals, as curcumin, resveratrol⁴⁵, genistein, and apigenin, can diminish cyclin D1 overexpression in malignancy cells ^{46, 47, 48}.

Shockingly, the information indicated that TAE fundamentally advances cell harm, initiates caspase 3, and lifts Bax/Bcl2 proportion. Also, the statement of Cyclin D1 altogether diminishes in TAE-treated MCF-7 cells. Along these lines, this plant can be presented as a contender for more investigation in disease treatment.

T.ammi has polyphenols that might be liable for its watched anticancer impact in this examination. Thymol (50-55%) is the most dynamic parts of T. ammi. It has been demonstrated that thymol has antiproliferative properties on non-little cell lung malignant growth cells, A549, interminable myeloid leukemia cells, K562, Hep-2 cells, murine B16 melanoma cells, and human metastatic bosom disease cells, MCF-7 ^49-50.

As of late, it has been accounted for that the anticancer impacts of thymol in metastatic bosom malignancy cells (MCF-7) depended on the initiation of the old style apoptosis reaction, remembering decline for mitochondrial layer potential and increment in cytochrome c discharge from mitochondria, decline in Bcl-2/Bax proportion, increment in caspase action, and cleavage of PARP and fracture of DNA, which have a place with the mitochondrial pathway of the apoptosis ⁵¹.

In the current examination, we assessed thymol-instigated MCF-7 harmfulness. The information indicated that TAE potentiates the cytotoxicity of thymol which is joined by an expansion in severed caspase 3 and an abatement in Cyclin D1 protein articulation. In any case, the point by point instruments of thyme extricate stay indistinct and ought to be explained by further examination.

CONCLUSION:

Taken together, this examination shows that T. ammi separate has a potential ant proliferative/proapoptotic property in MCF-7 cells and can be utilized as pharmaceutics contextual analysis for bosom malignant growth medications.

ACKNOWLEDGEMENT:

· I would like to thank Dr. Ajay Kumar Gupta, Dr. Nisha Sharma, Dr. A. Rajendrian, Dr. Ajay Yadav, Dr. Anju Singh, Dr. Pratima Katiyar of Institute of Pharmacy, CSJM University for their guidance and support.

· I would like to acknowledge Mr. Sagar Singh Jough, Asst. Professor, CSSGI, Etawah, for his guidance.

· Finally I would like to thank our family, friends and god for their cooperation and help.

CONFLICT OF INTEREST:

The author declare no conflict of interest.

REFERENCES:

1. B. Edwin Jose, P. Muralidharan. Effect of Azima tetracantha Lam on Human Breast Cancer Cells MCF -7 Research J. Science and Tech. 2019; 11(2):109-112.

2. Enayatrad, M., Amoori, N. & Salehiniya, H. Epidemiology and trends in breast cancer mortality in Iran. Iranian journal of public health 44, 430 (2015).

3. Maruthanila, V et al., J. Recent development of targeted approaches for the treatment of breast cancer. Breast Cancer 24, 191 (2017).

4. Emami, S. A. et al. Growth Inhibition and Apoptosis Induction of Essential Oils and Extracts of Nepeta cataria L on Human Prostatic and Breast Cancer Cell Lines. Asian Pacific journal of cancer prevention 17, 125 (2016).

5. Lahlou, M. The success of natural products in drug discovery. Pharmacol Pharm 4, 17 (2013).

6. Roy, A., Attre, T. & Bharadvaja, N. Anticancer agent from medicinal plants: a review. New apects in medicinal plants and pharmacognosy 1, 54 (2017).

7. "Oldest evidence of breast cancer found in Egyptian skeleton". Reuters. 24 March 2015. Archived from the original on 27 March 2015. Retrieved 25 March 2015.

8. "The History of Cancer". American Cancer Society. 25 March 2002. Archived from the original on 9 October 2006. Retrieved 9 October 2006.

9. R D, Ahmad A et al., A New Profile of Therapeutic Potential and Toxicities of Antineoplastic Drugs. Research J. Pharmacology and Pharmacodynamics. 2010; 2(6): 370-375.

10. Alfredo Morabia (2004). A History of Epidemiologic Methods and Concepts. Boston: Birkhauser. pp. 301–302. ISBN 978-3-7643-6818-0. Retrieved 31 December 2007.

11. Jump up to: a b Sulik, Gayle A. (2010). Pink Ribbon Blues: How Breast Cancer Culture Undermines Women's Health. USA: Oxford University Press. pp. 200–3. ISBN 978-0-19-974045-1. OCLC 535493589

12. Ferlay J, et al. Estimated Cancer Incidence, Mortality and Prevalance Worldwide in 2012. 2012. v1.0 (IARC CancerBase No. 11).

13. Global Burden of Disease Cancer Collaboration, The global burden of cancer 2013. JAMA Oncol 2015. [PMC free article] [PubMed]

14. Jemal A, et al., Global cancer statistics. CA Cancer J Clin. 2011; 61:69–90. [PubMed] [Google Scholar]

15. National Cancer Registry Programme. National Centre for Disease Informatics and Research. and Indian Council of Medical Research., Three year report of population based cancer registries 2009–2011 national cancer registry programme. National Cancer Registry, 2013.

16. Sharma K., Costas A., Shulman L.N., Meara J.G. A systematic review of barriers to breast cancer care in developing countries resulting in delayed patient presentation. J Oncol. 2012; 2012:8.

17. N. Moualla, M. Naser. Using GC/MS to Study the Chemical Composition of Essential Oil of Thymus vulgaris L. at AL-Qadmous Area, Syria. Research J. Pharm. and Tech. 8(4): April, 2015; Page 437-442.

18. Anjali Soni, Patel Femida, Preeti Sharma. In-vitro Cytotoxic Activity of Plant Saponin Extracts on Breast Cancer Cell-Line. Res. J. Pharmacognosy and Phytochem. 2017; 9(1): 17-22.2011; 3(1): 41-44.

19. Sankaradoss Nirmala, et al., Investigation of In vitro Anti-cancer property of Adhatoda vasica in Hela, HepG2, MCF-7, MDAMB-231 Cell Lines. Res. J. Pharmacognosy and Phytochem. 2019; 11(4):212-216. Dubey

20. Marchese, A., Orhan, I. E., Daglia, M., Barbieri, R., Di Lorenzo, A., et al. (2016) Antibacterial and antifungal activities of thymol: a brief review of the literature. Food Chem. 210, 402–414.

21. Xue, J., Davidson, P. M., and Zhong, Q. (2016) Inhibition of Escherichia coli O157: H7and Listeria monocytognes growth in milk and cantaloupe juice by thymol nanoemulsions prepared with gelatin and lecithin. Food Contr. 73, 1499–1506.

22. A.K Meena, et al., Comparative Studies on Quality Assessment of Trachyspermum Ammi Linn. Seeds Collected from Different locations of Punjab State. Research J. Pharmacognosy and Phytochemistry

23. Novy, P., Davidova, H., Serrano Rojero, C. S., Rondevaldova, J., Pulkrabek, J., et al. (2015) Composition and antimicrobial activity of Euphrasia rostkoviana Hayne essential oil. Evid. Based Complement. Altern. Med. 734101. https://doi.org/10.1155/2015/734101.

24. Sobczak, M., Kalemba, D., Ferenc, B., and Zylinska, L. (2014) Limited protective properties of thymol and thyme oil on differentiated PC12 cells with downregulated Mgst1. J. Appl. Biomed. 12, 235–243.

25. Güvenç, M., Cellat, M., Gökçek, _ I., Yavas¸, _ I., and Yurdagül Özsoy, Ş. (2018) Effects of thymol and carvacrol on sperm quality and oxidant/antioxidant balance in rats. Arch. Physiol. Biochem. 25, 1–8.

26. N. V. S. Venugopal, V. Nageswara Raoand, B. Sumalatha. Spectrophotometric Determination of Naproxen Sodium with Bromocresol Green and Bromothymol Blue as Chromogenic Reagents. Asian J. Research Chem. 4(5): May, 2011; Page 715-718.

27. Rakesh K. Joshi. Essential oil composition of Thymus linearis (Benth) from western Himalaya of Uttrakhand, India. Asian J. Pharm. Tech. 2016; 6(4): 199-201.

28. Kang, S.–. H., Kim, Y.–. S., Kim, E.–. K., Hwang, J.–. W., and Jeong, J.–. H. (2016) Anticancer effect of thymol on AGS human gastric carcinoma cells. J. Microbiol. Biotechnol. 26, 28–37.

29. Deepak Kumar Basedia, Birendra Shrivastava, B. K. Dubey, Pankaj Sharma. Synthesis and Anticancer Activity of Novel Substituted 1, 3, 4-oxadiazolo-[3, 2-a]-1, 3, 5-triazine and 1, 3, 4-thiadiazolo-[3, 2-a]-1, 3, 5-triazine Derivatives. Asian J. Research Chem. 7(3): March 2014; Page 310-315.

30. Sharifi‐Rad, M., Nazaruk, J., Polito, L., Morais‐Braga, M. F. B., Rocha, J. E., et al. (2018) Matricariagenus as a source of antimicrobial agents: from farm to pharmacy and food applications. Microbiol. Res. 215, 76– 88.

31. Satooka, H., and Kubo, I. (2012) Effects of thymol on B16‐F10 melanoma cells. J. Agric. Food Chem. 60, 2746– 2752.

32. Shettigar, N. B., Das, S., Rao, N. B., and Rao, S. B. (2015) Thymol, a monoterpene phenolic derivative of cymene, abrogates mercury‐induced oxidative stress resultant cytotoxicity and genotoxicity in hepatocarcinoma cells. Environ. Toxicol. 30, 968– 980.

33. Mendes‐da‐Silva, R. F., Lopes‐de‐Morais, A. A., Bandim‐da‐Silva, M. E., Cavalcanti, Gde., A., Rodrigues, A. R., et al. (2014) Prooxidant versus antioxidant brain action of ascorbic acid in well‐nourished and malnourished rats as a function of dose: a cortical spreading depression and malondialdehyde analysis. Neuropharmacology 86, 155– 160.

34. Günes‐Bayir, A., Kocyigit, A., Güler, E. M., and Kiziltan, H. S. (in press) Effects of thymol, a natural phenolic compound, on human gastric adenocarcinoma cells in vitro. Altern Ther. Health Med. pii: AT5782.

35. Vichai V, Kirtikara K (2006) Sulforhodamine B colorimetric assay for cytotoxicity screening. Nature protocols 1: 1112-1116.

36. M Lavanya, et al., Evaluation of Anticancer Activity of Ethanolic and Ethylacetoacetate Extracts of Sweet Cherry Against Human Breast Cancer Cell Line MCF-7. Research J. Pharmacology & Pharmacodynamics. 2016; 8(2): 65-70

37. B. Wolf, M. Brischwein, V. Lob, J. Ressler, and J. Wiest, “Cellular signaling: aspects for tumor diagnosis and therapy,” Biomedizinische Technik, vol. 52, no. 1, pp. 164–168, 2007.

38. J. J. Lu, Y. Y. Dang, M. Huang, W. S. Xu, X. P. Chen, and Y. T. Wang, “Anti-cancer properties of terpenoids isolated from Rhizoma Curcumae—a review,” Journal of Ethnopharmacology, vol. 143, no. 2, pp. 406–411, 2012.

39. A. M. Hunter, E. C. LaCasse, and R. G. Korneluk, “The inhibitors of apoptosis (IAPs) as cancer targets,” Apoptosis, vol. 12, no. 9, pp. 1543–1568, 2007.

40. C. C. Wu and S. B. Bratton, “Regulation of the intrinsic apoptosis pathway by reactive oxygen species,” Antioxidants and Redox Signaling, vol. 19, no. 6, pp. 546–558, 2013.

41. V. Baldin, et al., “Cyclin D1 is a nuclear protein required for cell cycle progression in G1,” Genes and Development, vol. 7, no. 5, pp. 812–821, 1993.

42. J. Adelaide, G. Monges, C. Derderian, J.-F. Seitz, and D. Birnbaum, “Oesophageal cancer and amplification of the human cyclin D gene CCND1/PRAD1,” British Journal of Cancer, vol. 71, no. 1, pp. 64–68, 1995.

43. L. M. Gumbiner, P. H. Gumerlock, P. C. Mack et al., “Overexpression of cyclin D1 is rare in human prostate carcinoma, ” Prostate, vol. 38, no. 1, pp. 40–45, 1999.

44. E. Peurala, et al., “The prognostic significance and value of cyclin D1, CDK4 and p16 in human breast cancer,” Breast Cancer Research, vol. 15, no. 1, article R5, 2013.

45. B. B. Aggarwal, et al., “Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies,” Anticancer Research, vol. 24, no. 5, pp. 2783–2840, 2004.

46. S. Shukla and S. Gupta, “Apigenin-induced cell cycle arrest is mediated by modulation of MAPK, PI3K-Akt, and loss of cyclin D1 associated retinoblastoma dephosphorylation in human prostate cancer cells,” Cell Cycle, vol. 6, no. 9, pp. 1102–1114, 2007.

47. A. Shehzad, F. Wahid, and Y. S. Lee, “Curcumin in cancer chemoprevention: molecular targets, pharmacokinetics, bioavailability, and clinical trials,” Archiv der Pharmazie, vol. 343, no. 9, pp. 489–499, 2010.

48. K. A. Hwang, et al., “Genistein, a soyphytoestrogen, prevents the growth of BG-1 ovarian cancer cells induced by 17β-estradiol or bisphenol A via the inhibition of cell cycle progression,” International Journal of Oncology, vol. 42, no. 2, pp. 733–740, 2013.

49. A. T. Koparal and M. Zeytinoglu, “Effects of carvacrol on a human non-small cell lung cancer (NSCLC) cell line, A549,” Cytotechnology, vol. 43, no. 1–3, pp. 149–154, 2003

50. K. M. Arunasree, “Anti-proliferative effects of carvacrol on a human metastatic breast cancer cell line, MDA-MB 231,” Phytomedicine, vol. 17, no. 8-9, pp. 581–588, 2010.

51. S. Karkabounas, O. K. Kostoula, T. Daskalou et al., “Anticarcinogenic and antiplatelet effects of carvacrol,” Experimental Oncology, vol. 28, no. 2, pp. 121–125, 2006.

Received on 21.04.2020 Modified on 23.05.2020

Asian J. Research Chem. 2020; 13(5):327-333.

DOI: 10.5958/0974-4150.2020.00063.2