Johnson Grace, Kharel Pranil, Goutham Matta

Culinary Herbs & Spices and Anti-Carcinogenic Properties

Herbs: Herbs are the fresh and dried leaves generally of temperate plants.

Generally herbs are green in color and have a pleasant taste.

Spices: Spices are the flowers, fruits, seeds, bark, and roots typically of tropical plants. Spices are brown, black or red in color with pungent smell.

Introduction

Brief History:Spices and herbs are woven in the history of mankind since the early civilization.

Introduction

Early records indicate that herbs and spices were used as medicinals in ancient Egypt and as food preservatives in ancient Rome and Greece.

Herbs and spices continued to be used during the Middle Ages for flavoring, food preservation, and/or medicinal purposes.Today, the business of producing herbs and spices has become one of the fastest growing industries in agriculture.

5 Years Growth: Herbs & Spices

Source: Nielsen Answers on Demand

Top 3 Sellers: Herbs & Spices

Source: Nielsen Answers on Demand

Source: againasttheodds.eu

Source: againasttheodds.eu

Source: againasttheodds.eu

Source: learnaboutcancer.com

According to the Global Oncology Trend Report, released by the IMS Institute for Healthcare Informatics, global spending on cancer medications rose 10.3 percent in 2014 to $100 billion, up from $75 billion in 2010.

The report follows one from IMS Health, which found that overall U.S. spending on drugs reached $373.9 billion in 2014 – a record high.

Introduction: Outgoings on drugs for cancer in U.S.A

Fighting cancer is something our body is doing all the time, not just when one is diagnosed with the disease.  

Some culinary herbs and spices are particularly rich in cancer preventive and cancer-fighting substances.

Herbs and spices serve as inhibitors of carcinogen bio-activation, decrease free radical formation, suppress cell division and promote apoptosis in cancerous cells.

Dragland et al. 2003 suggested that the addition of about 1 g/day of herbs and spices to one’s diet can significantly contribute to total antioxidant intake (>1 mmol) and offers a better source of antioxidants than many food items.

5 CANCER-FIGHTINGHERBS

BASIL

PARSLEY

RED CLOVER

ROSEMARY

CHICKWEED

Mechanism of action

Spices and herbs may be a key to determine the balance between pro- and anticancer factors that regulate risk and tumor behavior. 

Multiple factors may influence the need for spices for reducing the risk of cancer or changing the biological behavior of cancer cells.

Mechanism of action: Herbs & Spices

AntimicrobialAntioxidant

Anti-inflammatoryMediators

Anti-carcinogenicAnti-tumorigenic

Harmful MicrobesOxidation

Pro inflammatory- Mediators

CarcinogensTumorigenic-

Compounds

Low ------------------ Cancer ---------------- High

Dietary factors including herbs and spices

Herbs, Spices and Cancer

The selective cytotoxic anti-cancer properties and proteomic analysis

of Trigonella Foenum-GraecumAbdulaziz Alsemari,Fahad Alkhodairy,Ahmad Aldakan,Mai Al-Mohanna,

Eman Bahoush,Zakia Shinwari andAyodele Alaiya(2014)

Terms to Understand Trigonella foenum graecum: Fenugreek T-cell lymphoma: Lymphoma is the most common blood

cancer. Lymphoma occurs when cells of the immune system called lymphocytes, a type of white blood cell, grow and multiply uncontrollably. The body has two main types of lymphocytes that can develop into lymphomas: B-lymphocytes (B-cells) and T-lymphocytes (T-cells).

Proteomics: The study of proteins and their functions. Apoptosis: The death of cells that occurs as a normal and

controlled part of an organism's growth or development.

The current in vitro studies were prompted by the clinical profile of a previously reported case of primary CNS(Cerebral) T cell lymphoma.

Brain MRI with contrast was performed and revealed numerous cerebellar- enhancing lesions.

The patient initially went into clinical remission for 37 months after chemother-apy and radiotherapy.

After three years of remission, the patient’s disease relapsed with intermittent weakness in her left arm and deviation of her mouth to the left.

Background

The family decided to treat their child with concentrated extract of fenugreek, boiled in regular water. The approximate daily treatment amount was 8 g of fenugreek seeds, over the course of six months. 

Repeat brain MRI three months later showed significant improvement in the intra cerebral lesion.

The subsequent brain MRI after six months showed complete resolution of the relapsed lesion.

In this research demonstrated the selective cytotoxic effects of fenugreek extract in vitro on a panel of cancer cell lines, and a proteomics analysis of fenugreek.

Background

Background

Brain MRI shows enhanced lesion in the front parietal lesion.

Brain MRI with contrast shows complete resolution of relapsed lymphoma.

Research Questions Does fenugreek has selective cytotoxic effects against cancer

cells? Does fenugreek selectively induces apoptosis in cancer and

normal cells? Is there a significant difference between protein patterns of four

different regional fenugreek samples?

Summary of Research

Cell culture:T-cell lymphoma (TCP), B-cell lymphomas, Thyroid Papillary carcinoma

(FRO) and human breast cancer (MCF7)

Cells were cultivated at 37°C and 5% CO2 in Dulbecco’s MEM (DMEM) containing 5% fetal bovine serum, 3 mM glutamine and antibiotics

Fenugreek extraction:Fenugreek seeds were extracted in water, filtered, concentrated and

reconstituted to a working stock concentration of 50 mg/ml

Cells were exposed to fenugreek extract at different concentrations (100 ug/ml, 200 ug/ml and 300 ug/ml) and at

different time points (0, 24, 48, 72 and 96 hrs).

Lymphoma

Does fenugreek has selective cytotoxic effects against cancer cells?

Summary of Research

Cells were seeded onto a 96-well plate at a density of 5000 cells/well.

Incubated at 37°C for different time points (0, 24, 48, 72 and 96 hrs.) in a titrated medium containing different concentrations (100 ug/ml, 200 ug/ml and 300 ug/ml) of fenugreek extract.

96-Well Plate

Does fenugreek selectively induces apoptosis in cancer and normal cells?

Summary of Research: Research Question II

Approximately 5 × 105 cells were seeded and cultured in 60 mm plates.

Incubated at 37°C for 72 hours in a titrated medium containing different concentrations of fenugreek extract.

Thereafter, cells were stained with Propidium Iodide (PI) and Annexin V, using Vybrant Apoptosis Assay kit.Flow cytometric graphs of T-cell lymphoma without fenugreek treatment (A) and when treated with 300 μg/ml of Fenugreek, for 24, and 72 hours, B and C respectively. 

Summary of Research: Research Question II

Is there a significant difference between protein patterns of four different regional fenugreek samples?

Summary of Research: Research Question III

Seeds from four different regions were grounded and protein was ex-tracted.

2-D Electrophoresis was used to determine the protein patterns.

Representative 2-DE gels derived from all the 4 different regional Fenugreek samples (A) : Fenugreek taken by the patient. (B, C and D): Fenugreeks form other regions.

Sample pairs Correlation

Group correlation among pairs of A and B 0.37 (n= 3 pairs)

Group correlation among pairs of A and D 0.41 (n= 3 pairs)

Group correlation among pairs of A and C 0.36 (n= 3 pairs)

Group correlation among pairs of B and D 0.86 (n= 3 pairs)

Group correlation among pairs of B and C 0.83 (n= 3 pairs)

Group correlation among pairs of D and C 0.88 (n= 3 pairs)

Summary of Research: Research Question III

Correlation analysis of pairs of 2-DE gels of fenugreek samples from four different regions A, B, C and D. Note the poor correlation between pairs of fenugreek from region A vs. B/C/D, compared with good correlation among pairs of D/C,B/D, C/B

First human case in which established malignant CNS cancer showed regression, then disappearing of the cancer lesion with daily use of fenugreek extract.

Analysis of fenugreek extract indicated the presence of several compounds with anticancer properties, including gingerol, cedrene, zingerone, vanillin and eugenol.

Biologically active agents in fenugreek may vary based on geographical environments.

However, these exciting results obviously warrant additional study, including further drug characterization and development.

Conclusion

FenugreekFenugreek

Inhibition of breast tumor growth and angiogenesis by a medicinal herb:

Ocimum gratissimumPratima Nangia-Makker, Larry Tait, Malathy P.V. Shekhar, Eduardo Palomino, Victor Hogan, Marie P. Piechocki, Tatsuyoshi Funasaka and

Avraham Raz (2007)

Terms to Understand OG: Ocimum gratissimum (holy basil) COX-2: enzyme responsible for inflammation MDA-MB-435 and MDA-MB-231: human breast

cancer cells EIII8 and DCIS.com: human preneoplastic breast

cancer cells BAMEC: bovine adrenal medulla endothelial cells

“Holy Basil” or “Tulsi”Anti-oxidant, anti-carcinogenic, radio-pro-

tective and free radical scavenging properties

No detailed studies on effects on human cancers

Active constituents thought to be responsible for medical action: Eugenol, Apigenin, and Ursolic Acid

Background

Basil

Purpose: To analyze the impact of Ocimum on tumor

growth, angiogenesis, and metastasis related properties

To compare the analysis of eugenol, apigenin, and ursolic acid

Summary

Research Questions: What are the effects of Ocimum on human breast

cancer? How do eugenol, apigenin, and ursolic acid

compare in terms of tumor cell migration and 3D tube formation?

Summary

MethodsAqueous extracts prepared from mature OG

leaves Iceberg lettuce as control, prepared similarly to

OG leaf extract3D growth of DCIS.com and EIII8 cellsMDA-MB-435 and MDA-MB-231 were gifted Iceberg lettuce as control, prepared similarly to

OG leaf extract

Assay/TestsCell viability, cell proliferation, anchorage indepen-dent growth, three-dimensional growth and tube formation assay, induction of COX-2 expression and western blot analysis, angiogenesis assay, tu-mor growth in nude mice, immunohistochemical analysis, TUNEL assay, high performance liquid chromatography, and statistical analysis

Figure I: Growth curve of MDA-MB-231 cells in presence of OG extract a) Range between 10 and 15,000b) Range between 15 and 20,000Figure II: Anchorage independent growth of MDA-MB-231 cells on soft agar in presence of OG extracta) Control, b) .1% OG extract, c) histogram representing colony forming efficiency at various concentrations of OG extract

Figure I: Effect of OG extract and components on tube formation of EIII8 cells.(a) control untreated; (b) 0.05% OG extract (w/v); (c) 0.1% OG extract (w/v); (d) 625 μM eugenol, (e) 10 μm ursolic acid; (f) 18 μm apigenin, (g) 0.01% ethanol; (h) 1.25 mM KOH.

International Journal of CancerVolume 121, Issue 4, pages 884-894, 16 APR 2007 DOI: 10.1002/ijc.22733http://onlinelibrary.wiley.com/doi/10.1002/ijc.22733/full#fig3

Aqueous extract of Ocimum leaves inhibit tumor growth and angiogenesis by Tumor cell proliferation Migration Morphogenesis Stromal apoptosis Induction of COX-2

Inhibitory properties not due to any 1 of 3 active components Due to undescribed component or combination of components?

Conclusion

Cinnamon extract induces tumor cell death through inhibition of

NFκB and AP1Ho-Keun Kwon, Ji-Sun Hwang, Jae-Seon So,Choong-Gu Lee, Anupama Sahoo, Jae-Ha Ryu, Won Kyung Jeon,Byoung Seob Ko,Chang-Rok Im, Sung Haeng Lee,

Zee Yong Park andSin-Hyeog Im (2010)

Terms to Understand NFkB: It is a protein complex which controls

transcription of DNA, cytokine production and cell survival.

AP1: It regulates gene expression in response to stimuli like cytokines, growth factors, stress & bacterial & viral infections. It controls cell growth, proliferation & apoptosis.

Cinnamomum cassia bark is the outer skin of a tree . Its extract contains essential oils (cinnamic aldehyde and cinnamyl aldehyde),

tannin, mucus and carbohydrates. It has got the several biological functions such as anti-oxidant, anti-microbial,

anti-inflammation, anti-diabetic effects and anti-tumor activity. Tumor cells are generally resistant to apoptosis. Thus, killing the tumor cells by

stimulating the pathway of apoptosis is the best way to develop anticancer agents.

NFkB and AP1 play important roles in tumor development. It is reported that anticancer effect of cinnamon extract is associated with

modulation of angiogenesis and effector function of CD8* T cells

Background

Research Questions:Does cinnamon extract inhibit tumor cell proliferation and

induced tumor cell death?Does cinnamon extract induces active cell death of

melanoma cells?Does cinnamon extract inhibits the melanoma growth by

inhibiting NFκB and AP1?

Summary

Summary: (In Vitro)Treatment of cinnamon extract inhibits the growth of various cancer cells. Cinnamon (0.5 mg/ml) was treated for 0, 48 and 72 hrs to cancer cell lines (Hela; cervical cancer, Caco2; colon cancer, Clone M3; melanoma and B16F10; melanoma). After treatment of cinnamon extract, (A) morphological changes of each cancer cell lines were monitored by microscopic observation. (B) Proliferation and viability of cancer cells were measured by cell viability assay at the indicated time points.

Oral administration of cinnamon extract significantly inhibits melanoma progression in vivo

Oral administration of cinnamon extract inhibits melanoma growth in vivo

Cinnamon extract treatment induces active cell death in melanoma cells. At the indicated time periods after treatment of cinnamon extract, cells were stained with Annexin V and 7-ADD. Double positive (Annexin V+ and 7-ADD+) (A) or Annexin V+ positive (B) populations were analyzed by FACS. Gene expression (C) and protein (D) level of pro-apoptotic molecules such as Bad, Bim, Bax and Bak were measured by quantitative real-time PCR or immunoblotting, respectively.

Cinnamon extract treatment down-regulates the levels of NFκB and AP1 and their target genes

Anti-tumor effect of cinnamon extract is linked to the re-duced levels of NFκB and AP1 in vivo melanoma model

Cinnamon extract treatment induces tumor apoptosis by decreasing the levels of NFκB and AP1 and their target genes.

The extracts of cinnamon inhibited the various tumor cell growths in vitro and suppressed in vivo melanoma progression.

The cinnamon extract mediate the anti-cancer effect by apoptosis induction and NFkB & AP1 blockade.

Conclusion

Compare and Contrast of Research Articles

Article Main Finding Strengths Weaknesses Future Studies

Impact of Ocimum on Breast Cancer Tumor Growth

Aqueous extract of Oci-mum leaves inhibit tumor growth and angiogenesis

Inhibitory properties not due to any 1 of 3 active components

Comparing the analysis of active components: eugenol, apigenin, and ursolic acid

Use of human breast cancer cells 

Individual active compo-nents had no affect

Isolate active compo-nents

The selective cytotoxic anti-cancer properties and proteomic analysis of Trigonella Foenum-Graecum

Fenugreek has selective cytotoxic effects against cancer cells.

Observed a significant degree of heterogeneity in the protein expression between samples

First human study to show anticancer proper-ties of fenugreek in pa-tient with relapsed CNS-lymphoma

Comparing the protein content between fenu-greek samples

Geographical Locations of samples were not specified.

Limited sample size.

A more rigorous scientific evaluation, such as a clinical trial, is obviously warranted before the anti cancer properties of fenugreek can be con-firmed.

Cinnamon extract in-duces tumor cell death through inhibition of NFkB and AP1

Extracts of cinnamon inhibit the tumor cell growths in vitro and suppressed in vivo melanoma progression.

The extract of cinnamon mediates the anti-cancer effect by apoptosis induction and NFkB and AP1 blockade.

Use of human cancer-ous cells.

The temperature of wa-ter used for extraction is not mentioned.

The human melanoma cells are not used.

Further study on hu-man melanoma cells can be done.

Mechanism of action of cinnamon can be studied.

Future ResearchDNA

RepairCarcinogen Metabolism

Inflammatory Response

CarcinogenMetabolism

HormonalRegulation

DifferentiationApoptosis

CellCycle

Future Research Currently available data are intriguing, the effective exposures needed to

bring about the desired outcome(s), and what interactions (both positive and negative) exist with other components of the diet or with medications that an individual may regularly consume

Clarification is also needed regarding which types of cells respond to various herbs, spices, and their bioactive components and further exploration of the potential for gender-based differences in response.

References:Dragland S, Senoo H, Wake K, Holte K, Blomhoff R. Several culinary and medicinal herbs are important sources of dietary antioxidants. J Nutr. 2003;133:1286–90.

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