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A REVIEW ON ANTIPROLIFERATIVE ACTIVITY OF PLANT EXTRACTS AGAINST BREAST CANCER CELL LINES

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A REVIEW ON ANTIPROLIFERATIVE ACTIVITY OF PLANT EXTRACTS AGAINST BREAST CANCER CELL LINES

R. Kumar * 1, S. Mahey 2, V. Kumar 1, R. Arora 3, A. Sharma 4 and S. Arora 5

Department of Botany 1, DAV University, Jalandhar - 144012, Punjab, India.

Department of Botany 2, DAV College, Jalandhar - 144001, Punjab, India.

Department of Biochemistry 3, Sri Guru Ram Das Institute of Medical Sciences and Research, Amritsar - 143501, Punjab, India.

State Key Laboratory of Subtropical Silviculture 4, Zhejiang A and F University, Hangzhou - 311300, China.

Department of Botanical and Environmental Sciences 5, Guru Nanak Dev University, Amritsar - 143005, Punjab, India.

ABSTRACT: Breast cancer (BC) is the foremost cause of deaths among women worldwide. Plants contain diverse bioactive phytochemicals which have been explored by researchers all over the world for their cancer preventive potential. The present review tabulates in-vitro tested plants during 2014-17 against breast cancer cell line. We have collected data of 56 angiosperm families (117 plant species) which was subjected to cluster analysis. On the bases of IC50 values of plant extracts were clustered using cluster analysis Cluster analysis showed a grouping of order Brassicales, Fabales, Lamiales, Caryophyllales, Myrtales, and Apiales. It has been found that most of the plants tested against BC belong to eudicot group of plants. Active plant extract obtained after 24 h, 48 h and 72 h treatment were Mimosa caesalpinnifolia, Ferulago angulat, Magydaris tomentosa, and Ipomea batatas respectively. These plants may further be characterized for active ingredients to check their prospects in breast cancer treatment.

Keywords:

MTT, Breast cancer, Cluster analysis, MCF-7

INTRODUCTION: Breast Cancer (BC) is the most prevalent cause of cancer-related deaths among women worldwide 1, 2, 3, 4, 5. Despite advances in its diagnosis and treatment options, the number of incidences is increasing every year 6, 7. The number of breast cancer cases reported in 2018 from the whole world was 20,88,849 amongst which 6,26,679 died 8. BC is not just one disease but has many variations and subtypes with distinct signatures and treatment programs 9, 10.

The early stage BC can be successfully cured, but treatment options are scarcely accessible to patients with advanced or metastatic stages 7. Women with mutated BRCA gene, have nearly 80% risk of developing BC along with a 50% possibility of their children getting the mutated gene 11. The genetic mutations in BRCA1 and BRCA2 genes suppurates the exigent root cause of patrimonial breast cancer 12. Although numerous causes are associated with the establishment and progression of BC, yet the oxidative stress (OS) is operating in most of the intracellular pathways concerned with cellular proliferation 13.

It has been reported repeatedly that the level of OS is higher for BC patients as compared to healthy people due to genetic abnormalities 1, 14. This elevated OS is beneficial for malignant cells as it upregulates the reactive oxygen species (ROS) mediated signaling pathways which encourage cell growth, cell differentiation, glucose synthesis, protein synthesis and hence cell survival. Numerous ROS are generated within the body as a result of basal metabolic activities. Hydrogen peroxide (H2O2) is one such ROS which is generated during estrogen metabolism, and it is known to activate extracellular regulated kinase 1/2 (Erk 1/2). Erk 1/2 is known to promote the survival of breast cancer cells in humans by activating downstream elements 15. Plant extracts being excellent scavengers of free radicals have often been implicated as a remedial measure in various diseases. The plant extracts contain numerous phytochemicals which act synergistically against disorders unlikely the purified compounds 16. Numerous well established anticancer drugs in use today have been derived from plants such as Sulphoraphane, Paclitaxel, Epipodophyllotoxin, Vincristine, Vinblastin, Vinorelbine, Vindesine, Vinflunine, Pomiferin, Roscovitine, Flavopiridol, Noscapine 17, 18, 19. In light of the significant contribution of phytochemicals in cancer treatment, the present review was designed to compile the in-vitro antiproliferative activity of various plant extracts against breast cancer cell line.

2. MATERIALS AND METHODS:

2.1. Database Search: We have searched online free resource “PubMed” (maintained by National Center for Biotechnology Information at the National Library of Medicine, USA) for plants extracts assessed for their antiproliferative activity against breast cancer cell line (MCF-7) employing in-vitro assays (MTT, MTS, XTT, CCK-8, SRB, CVS, WST-1, ATPlite, Alamar blue, Methylene blue, RTCA MP) for treatment period of 24, 48 and 72 h. The data was collected for the last four years i.e, 2014-17.

2.2. Presentation of Collected Data: The collected data was presented in the tabulated form. Various parameters selected for the present work were a plant, family, plant part used, the solvent used for extraction, assay employed and IC50 concentration.

2.3. Data Analysis: Cluster analysis was done by using PAST software applying Ward’s method, and Euclidian distance was calculated and presented as a measure of similarity.

3. RESULTS AND DISCUSSION:

3.1. Families and Orders: The collected data covered 56angiosperm families and 118 plants Table 1. From these, 50 families belong to eudicots (22 orders), 4 families belong to monocots (4 orders), and 2 families belong to magnoliids (2 orders) as shown in Table 2.

3.2. Cluster Analysis (CA): The CA was applied to IC50 values of different plant species (as reported in the respective research paper) on the basis of above-ground plant part with treatment time of 24 (AG24), 48 (AG48) & 72 h (AG72); below ground with treatment period of 24 h (BG24).

TABLE 1: PLANT SPECIES TESTED AGAINST MCF-7 BREAST CANCER CELL LINE DURING 2014-18

S. no. Family Plant Plant part Solvent Assay Treatment time (H) IC50 (µg/ml)
1 Acanthaceae Clinacanthus nutans 32 Root MeOH MTT 24 35
Avicennia alba33 Leaves MeOH MTT 48 117
Ecbolium virde33 Leaves MeOH MTT 48 60
Phlogacanthus thyrsiformis34 Leaves EtOH MTT 24 49
2 Anacardiaceae Mangifera indica35 Kernal EtOH MTT 72 15
Lannea coromandelica33 Bark MeOH MTT 48 270
Leaves MeOH MTT 48 161
3 Apiaceae Glehnia littoralis36 Root AQ CCK-8 24 158.63
Ferula heuffelii37 Underground Parts CF MTT 24 6.1
MeOH MTT 24 47.6
4 Apocynaceae Leptadenia reticulata38 Whole plant AQ MTT 24 740
Picralima nitida39 Root Bark EtOH MTS 24 22.76
Holarrhena floribunda40 Leaves MeOH MTT 24 357.6
48 244.3
72 126.7
Hemidesmus indicus41 Root AQ MTT 24 829.32
30% EtOH MTT 24 209.73
5 Araliaceae Hedera nepalensis42 Aerial Part MeOH+ CF SRB 72 62
6 Asteraceae Vernonia cinerea43 Whole plant EtOH SRB 72 60
Gnaphalium luteoalbum33 Leaves MeOH MTT 48 340
Artemisia indica34 Leaves 95% EtOH MTT 24 48
Launaea procumbens44 Leaves MeOH MTT 48 ˃ 80
Sphaeranthus amaranthoide45 Whole plant PE MTT 48 23.55
Anthemis mirheydari46 Whole plant DCE MTT 72 25.2
7 Berberidaceae Berberis orthobotrys39 Root MeOH MTS 24 81.35
8 Bignoneaceae Tabebuia impetigosa47 Dried inner bark MeOH SRB 24 110.76
9 Cactaceae Opuntia ficus-indica48 Stem EA MTT 48 138
10 Capparaceae Crateva adansonii49 Stem Bark DCM + MeOH ABA 24 289
11 Caprifoliaceae Nardostachys jatamansi50 Roots + Rhizome MeOH MTT 48 58.01
PE MTT 48 60.59
12 Caryophyllaceae Arenaria montana51 Aerial part AQ SRB 48 130.05
13 Chrysobalanaceae Licania rigida52 Seeds EtOH ATPlite 24 N.C
Licania tomentosa52 Seeds EtOH ATPlite 24 N.C
14 Combretaceae Anogeissus latifolia53 Stem & Leaves 95% EtOH MTT 48 20.1
Terminalia bellerica53 Stem & Bark 95% EtOH MTT 48 9
Terminalia bellerica54 Fruits 70% MeOH WST-1 48 104.65
Terminalia chebula55 Leaf galls EtOH MTT 72 208.16
15 Convolvulaceae Ipomoea batatas56 Peeled Root tubers Acidified EtOH MTT 24 5.9
Not Peeled Root tubers Acidified EtOH MTT 24 4.9
16 Cucurbitaceae Momordica cochinchinensis57 Aril Hexane+ Acetone + EtOH MTT 48 117
17 Dilleniaceae Dillenia suffruticosa58 Root DCE MTT 24 20.3
Dillenia suffruticosa59 Root EA MTT 24 76
Dillenia indica33 Leaves MeOH MTT 48 340
18 Dioscoreaceae Dioscorea villosa60 Root MeOH CC 72 27
19 Dipterocarpaceae Dipterocarpus turbinatus33 Bark MeOH MTT 48 168
20 Droseraceae Drsera burmanni61 Whole plant MeOH WST-1 48 120.94
21 Ebenaceae Diospyros peregrina33 Leaves MeOH MTT 48 7
22 Ericaceae Vaccinium bracteatum62 Leaves Hexane RTCA MP 72 206.75
23 Euphorbiaceae Jatropha curcas39 Root Bark EtOH MTS 24 36.55
Jatropha gossypifolia39 Root Bark EtOH MTS 24 25.55
Croton sphaerogynus63 Leaves EtOH SRB 48 53.4
24 Fabaceae Eythrina excelsa64 Stem Bark EtOH MTT 24 13.6
Acacia catechu53 Fruit 50% EtOH MTT 48 22.8
Acacia catechu54 Heartwood 70% MtOH MTT 48 288.85
Enterolobium cyclocarpum65 Leaves MeOH MTT 48 11.84
Sophora interrupta66 Root EA MTT 24 250
Caesalpinia pulcherrima33 Leaf MeOH MTT 48 240
Clitoria ternatea33 Flower MeOH MTT 48 114
Leaf MeOH MTT 48 170
Alhagi graecorum67 Aerial Part 85% EtOH CVA 48 36.4
Saraca indica68 Bark 80% MtOH MTT 72 73.6
Pithecellobium dulce16 Leaf AQ MTT 24 400
48 300
Mimosa caesalpiniifolia69 Leaves 70% EtOH MTT 24 5
25 Geraniaceae Pelargonium sidoides70 Root AQ + Alcohol SRB 72 43
26 Hypericaceae Hypericum adenotrichum71 Aerial part MeOH MTT 72 10.9
27 Icacinaceae Pyrenacantha standtii72 Leaves EtOH MTS 24 37.36
28 Lamiaceae Teucrium polium73 Leaves MeOH MTT 48 35
Flower MeOH MTT 48 20
Lavandula dentata74 Whole plant EtOH MTT 24 39
Coridothymus capitatus75 Aerial part EtOH MTT 24 100
Lavandula angustifolia76 Aerial part Hexane MTS 24 85.68
EtOH MTS 24 179.6
Stachys acerosa77 Aerial part DCM MTT 72 160.3
Stachys benthamiana77 Aerial part DCM MTT 72 71.1
MeOH MTT 72 162.4
Stachys byzantina77 Aerial part DCM MTT 72 131
Stachys lavandulifolia77 Aerial part DCE MTT 72 81.2
80% MeOH MTT 72 151.7
Stachys persica77 Aerial part 80% MeOH MTT 72 104.1
Stachys pilifera77 Aerial part DCM MTT 72 40.9
Stachys pubescens77 Aerial part DCM MTT 72 103.3
MeOH MTT 72 146.5
Stachys spectabilis77 Aerial part DCM MTT 72 65.2
Melissa officinalis78 Leaves AQ MTT 48 51
Nepeta cataria79 Aerial part MeOH ABA 48 ˃ 500
Salvia chorassavica80 Root MeOH ABA 48 13
29 Lauraceae Cinnamomum cassia81 Bark Hexane MTT 24 34
30 Lecythidaceae Barringtonia racemosa82 Fruit MeOH MTT 48 57.61
31 Loranthaceae Plicosepalus curviflorus83 Leaves MeOH CVS 48 20.9
Macrosolen parasiticus84 Stem MeOH MTT 48 97.33
MeOH SRB 48 51.9
AQ MTT 48 59.33
AQ SRB 48 94.58
32 Malvaceae Theobroma cacao85 Leaf MeOH MTT 24 41.4
Bark MeOH MTT 24 72
Root MeOH MTT 24 76.4
Abutilon theophrasti86 Aerial Part MeOH MTT 24 505.8
Ceiba pentandra87 Stem Bark PE MTT 48 152.17
72 330.85
Hibiscus sabdariffa82 Fruit MeOH MTT 48 112.1
33 Melastomataceae Melastoma malabathricum88 Leaves MeOH MBA 72 7.14
Flower MeOH MBA 72 33.63
34 Molluginaceae Glinus oppositifolius33 Whole plant MeOH MTT 48 150
35 Moraceae Ficus cyathistipula89 Leaves Aqueous SRB 48 30
EtOH SRB 48 18
36 Moringaceae Moringa oliferna53 Leaves 50% EtOH MTT 48 26.4
Moringa oliferna90 Essential oil CP MTT 24 226.1
37 Myricaceae Myrica nagi33 Leaves MeOH MTT 48 172
38 Myrtaceae Syzygium aromaticum91 Cloves EtOH MTT 48 455
Pimenta dioica92 Berries AQ MTT 72 50
39 Oleaceae Fraxinus micrantha93 Dried Bark MeOH MTT 24 18.95
Jasminum sambac33 Leaves MeOH MTT 48 7
40 Papaveraceae Chelidonium majus94 Whole plant EtOH MTT 24 179.35
48 44.65
Fumaria vaillantii95 Aerial part 80% EtOH MTT 24 90
48 20
72 2
41 Phyllanthaceae Flueggea leucopyrus96 Aerial part AQ SRB 24 27.89
42 Piperaceae Piper cubeba97 Seeds MeH MTT 72 22.31
DCM MTT 72 62.2
Piper nigrum98 Seeds 93% EtOH MTT 24 27.1
43 Poaceae Cymbopogon citratus99 Leaves 50% EtOH MTT 48 68
90% EtOH MTT 48 104.6
44 Potenderiaceae Eichhornia crassipes100 Whole Plant MeOH SRB 72 1.2
45 Primulaceae Aegiceras corniculatum33 Fruit MeOH MTT 48 91
Maesa macrophylla34 Leaves 95% EtOH MTT 24 22.66
46 Punicaceae Punica granatum101 Fruit Fruit Juice MTT 72 50
47 Resedaceae Ochradenus arabicus74 Aerial part 95% EtOH MTT 24 562
48 Rhamnaceae Ziziphus spina-christi102 Leaves 80% EtOH MTT 24 230
49 Rubiaceae Hymenodictyon excelsum33 Bark MeOH MTT 48 80
Wood MeOH MTT 48 72
Mussaenda glabrata33 Leaves MeOH MTT 48 133
Galium aparine103 Whole plant MeOH XTT 72 503
50 Rutaceae Glycosmis pentaphylla104 Leaves PE SRB 48 95.5
51 Salicaceae Casearia sylvestris105 Leaves AQ EtOH MTT 24 141
52 Solanaceae Capsicum annuum106 Seeds AQ CPC 120 14.7
53 Thymelaeaceae Phaleria macrocarpa107 Fruit AQ MeOH MTT 24 96
Seed AQ MeOH MTT 24 12
54 Urticaceae Urtica pilulifera75 Aerial part EtOH MTT 24 63
55 Verbenaceae Lantana camara108 Whole plant EtOH MTT 24 32.39
Clerodendrum viscosum33 Leaves MeOH MTT 48 50
56 Zingiberaceae Curcuma zedoaria109 Rhizome Hexane MTT 72 18.4
Etlingera elatior110 Flowers Water + EtOH MTT 72 173.1
Alpinia galanga111 Rhizome EtOH MTT 72 170
Curcuma kwangsiensis112 Rhizome HD MTT 24 82.3

N.C = Not Cytotoxic; MeOH = Methanol; EtOH = Ethanol; DCM = Dichloromethane; DCE = Dichloroethane; PE = Petroleum ether; EA = Ethyl Acetate; ABA = Alamar Blue Assay; CC = Cell Counting; CVA = Crystal Violet Assay; MBA = Methylene Blue Assay; CPC = Coulter particle counter; AQ = Aqueous; CF = Chloroform; HD = Hydrodistillation; CP = Cold Pressing; CPC = Coulter particle counter; MBA = Methylene blue assay.

TABLE 2: NUMBER OF ORDERS AND FAMILIES OF PLANT SPECIES (TESTED AGAINST BREAST CANCER CELL LINE) BELONGING TO VARIOUS APG CLADES

S. no. APG Clade No. of orders No. of families No. of plants
1 Eudicots 22 50 108
2 Monocots 04 04 07
3 Magnoliids 02 02 03

3.2.1. CA of AG24: AG24 involved 31 plant species belonging to 23 families. The CA of AG24 revealed a clustering of Crateva adansonii (Capparaceae) with Moringa oliferna (Moringaceae) both belonging to order Brassicales and Mimosa caesalpiniifolia with Eythrina excelsa which belong to family Fabaceae and order Fabales Fig. 1. The minimum IC50 value was found in Momosa caesalpiniifolia (5.0 µg/ml) while maximum IC50 in Leptadenia reticulata (740 µg/ml).

3.2.2. CA of AG48: AG48 involved 44 plants belonging to 31 families. The CA of AG48 showed clustering of Clerodendrum viscosum (Verbenaceae) with Melissa officinalis (Lamiaceae), both belonging to order Lamiales. Clustering was also observed in plants of Caryophyllales order i.e, Opuntia ficus-indica (Cactaceae) with Arenaria montana (Caryophyllaceae) Fig. 2. The minimum IC50 value was found in Ferulago angulata (5.3 µg/ml) while maximum IC50 in Syzygium aromaticum (455 µg/ml).

3.2.3. CA of AG72: The AG72 group had 27 plants which were spread across 16 families. The CA presented aggregation of families belonging to order Lamiales and Myrtales. The plants belonging to order lamiales were Stachys persica, Stachys pubescens and Stachys byzantinai.

Order Myrtales included Pimenta dioica and Punica granatum Fig. 3. The minimum IC50 value was found in Magydaris tomentosa (0.94 µg/ml) while maximum IC50 in Galium aparins (503 µg/ml).

3.2.4. CA of BG24: The BG24 group included 13 plants belonging to 10 families. The CA showed grouping of Glehnia littotalis and Hemidesmus indicus belonging to family Apiaceae and order Apiales Fig. 4.

The minimum IC50 value was found in Ipomoea batatas (5.9 µg/ml) while maximum IC50 in Sophora interrupta (250 µg/ml).


3.3. Assay Reported: The in-vitro tetrazolium and resazurin-based reduction assays employed by authors were MTT (3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide); CCK-8 (Cell counting kit-8); MTS (3-(4,5-dimethylthiazol-2-yl)-5- (3-carboxymethoxyphenyl)- 2- (4-sulfophenyl)-2H-tetrazolium, inner salt); XTT (2,3-bis(2-methoxy-4-nitro-5-sulphophenyl)-5-carboxanilide-2H-tetrazolium, monosodium salt); SRB (Sulforhodamine B colorimetric assay); ABA (Alamar blue assay); WST (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt); Luminogenic ATP (Adenosine triphosphate); RTCA-MP (real-time cell impedance-based cell growth method); CVA (Crystal violet staining).

3.4. Causes of Breast Cancer: There are numerous causes of breast cancers as conversed in proceeding discussion. Elderly women are at more risk of developing breast cancer as compared to younger women 20. Post-menopausal women develop a greater threat of having breast cancer, which doubles with every passing decade to 80 years of life 21. The risk of getting breast cancer is reduced by bearing a child. This has been justified by the lower rates of incidences in married women as compared to single women 22. The reason may be early differentiation of mammary stem cells which belittles the threat of developing breast cancer 23.

Mutations in certain high penetrance genes like RCA1, BRCA2, PTEN, TP53, CDH1, and STK11and lower penetrance genes (CHEK2, BRIP1, ATM, and PALB2) are responsible for breast cancer incidences 24, 25. Long term or frequent exposure to polycyclic aromatic hydrocarbons (PAHs) disrupt estrogen metabolism and induce mammary cancer 26.

Most of the ovarian hormones taken after menopause to allay its effects increase the rate of breast cancer induction in postmenopausal women 27. Increased alcohol consumption in women is also linked with breast cancer incidences 28. Lanky lifestyle like persistent obesity and unhealthy dietary intake is associated with breast cancer 29.

Exposure to ionizing radiations especially during breast development elevates the risk of developing breast cancer which can be avoided by lessening repetitive needless testing 29, 30. Lopsided work schedule of women in developing as well as developed nations have also been positively correlated to increased breast cancer incidences 31. Table 3 and Fig. 5 compares the number of breast cancer cases reported their percentage of mortality in different countries.

TABLE 3: BREAST CANCER INCIDENCES REPORTED AND PERCENTAGE of CANCER DEATHS IN DIFFERENT COUNTRIES (GLOBOCAN 2014)

Country Population Breast cancer % of breast cancer death cases
India 124×107 144937 21.5
Bangladesh 155×106 14836 16.9
Afgganistan 29825000 3108 22.8
Australia 23050000 14710 17.3
China 139×107 187213 -
France 63937000 54245 19.9
Germany 82800000 71623 18.8
Italy 60885000 50658 18.2
Japan 1.27×106 55710 9.2
Pakistan 1.79×106 34038 30.8
South Africa 52386000 9815 16
Sri Lanka 21098000 3955 18.8
USA 318×106 232714 16.1

FIG. 5: NUMBER OF BREAST CANCER PATIENTS REPORTED AND PERCENTAGE CAUSALITIES IN DIFFERENT COUNTRIES

CONCLUSION: It has been concluded that most of the plants tested against breast cancer cell line belong to eudicots. Above ground plant parts showed better antiproliferative activity as compared to below ground plant parts. Active plant extract obtained after 24 h, 48 h and 72 h treatment were Mimosa caesalpinnifolia, Ferulago angulat, Magydaris tomentosa, and Ipomea batats. These plants must be characterized for active ingredients which can further be used in-vivo studies in animals induced with breast cancer to find out their prospects in breast cancer treatment.

ACKNOWLEDGEMENT: Authors thank DAV University, Jalandhar for providing necessary facilities for writing this review.

CONFLICT OF INTEREST: Authors declare no conflict of interest.

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    How to cite this article:

    Kumar R, Mahey S, Kumar V, Arora R, Sharma A and Arora S: A review on antiproliferative activity of plant extracts against breast cancer cell lines. Int J Pharm Sci & Res 2019; 10(7): 3144-54. doi: 10.13040/IJPSR.0975-8232.10(7).3144-54.

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Article Information

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English

IJPSR

R. Kumar *, S. Mahey, V. Kumar, R. Arora, A. Sharma and S. Arora

Department of Botany, DAV University, Jalandhar, Punjab, India.

raakysh@gmail.com

23 October 2018

13 February 2019

08 March 2019

10.13040/IJPSR.0975-8232.10(7).3144-54

01 July 2019

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