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Critical Reviews in Food Science and Nutrition

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Rosemary and Cancer Prevention: Preclinical

Perspectives
a

Suong N. T. Ngo , Desmond B. Williams & Richard J. Head

School of Animal and Veterinary Sciences , University of Adelaide , Roseworthy, SA,

Australia
b

School of Pharmacy and Medical Sciences and the Sansom Institute for Health Research ,
University of South Australia , SA, Australia
c

Preventative Health National Research Flagship CSIRO , Adelaide, SA, Australia

Accepted author version posted online: 01 Jun 2011.Published online: 26 May 2011.

To cite this article: Suong N. T. Ngo , Desmond B. Williams & Richard J. Head (2011) Rosemary and Cancer Prevention:
Preclinical Perspectives, Critical Reviews in Food Science and Nutrition, 51:10, 946-954, DOI: 10.1080/10408398.2010.490883
To link to this article: http://dx.doi.org/10.1080/10408398.2010.490883

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Critical Reviews in Food Science and Nutrition, 51:946954 (2011)

C Taylor and Francis Group, LLC
Copyright

ISSN: 1040-8398 print / 1549-7852 online
DOI: 10.1080/10408398.2010.490883

Rosemary and Cancer Prevention:

Preclinical Perspectives
SUONG N. T. NGO,1 DESMOND B. WILLIAMS,2 and RICHARD J. HEAD3
1

School of Animal and Veterinary Sciences, University of Adelaide, Roseworthy, SA, Australia
School of Pharmacy and Medical Sciences and the Sansom Institute for Health Research, University of South Australia,
SA, Australia
3
Preventative Health National Research Flagship CSIRO, Adelaide, SA, Australia

Downloaded by [189.200.34.179] at 07:28 19 January 2014

Colorectal cancer is the second leading cause of cancer death in Australia. Nutrition, particularly intake of vegetables
and certain plant components, has been reported to have a major role in cancer risk reduction. Recently, there has been a
growing research interest in rosemary, a common household plant grown in many parts of the world. This study aims to review
scientific evidence from all studies, published from 1996 to March 2010 that examined the protective effects of rosemary
on colorectal cancer and other types of cancer. Literature evidence from animal and cell culture studies demonstrates the
anticancer potential of rosemary extract, carnosol, carnosic acid, ursolic acid, and rosmarinic acid. No evidence for other
rosemary constituents was found. The reported anticancer properties were found to arise through the molecular changes in
the multiple-stage process of cancer development, which are dose related and not tissue or species specific. This is evidenced
by the ability of rosemary to suppress the development of tumors in several organs including the colon, breast, liver, stomach,
as well as melanoma and leukemia cells. The results suggested that the different molecular targets modulated by rosemary
and its active constituents are useful indicators of success in clinical cancer chemo-prevention trials.
Keywords carnosol, carnosic acid, ursolic acid, rosmarinic acid, anti-carcinogenesis

INTRODUCTION
Cancer is a main cause of morbidity and mortality in many
industrial countries. Colorectal cancer in particular is the most
common cancer reported to the Australian cancer registries.
Colorectal cancer is also a noticeable major health problem in
Australia and there are about 12,600 new cases and 4700 deaths
each year. Approximately one in 21 Australians is likely to
develop colorectal cancer during their lifetime. The risk is higher
after the age of 40 years and increasing progressively from 50
years of age (Stryker et al., 1987). The latest available Australian
national figures show that colorectal cancer was responsible for
13% of cancer deaths in 2001, with an estimated 29,058 lifeyears lost from premature death from colorectal cancer before
the age of 75 years (AIHW, 2010). Overall, colorectal cancer is
the third leading cause of death from cancer for both men and
women in the United States and Australia (ACS, 2010; AIHW,
2010) and the second leading cause of cancer death in Canada
(CCS, 2010).
Address correspondence to Suong N. T. Ngo, School of Animal and Veterinary Sciences, University of Adelaide, Roseworthy, SA 5371, Australia. Tel.:
(61) 8 8313 0660; Fax: (61) 8 8303 7956. E-mail: suong.ngo@adelaide.edu.au

Most colorectal cancers are believed to develop from slowly

growing benign polyps (Hill et al., 1978; Stryker et al., 1987).
Environmental factors have been found to greatly influence the
multiple-stage process which leads from the precursor lesion
to cancer (Potter et al., 1993; Winawer et al., 1993). In the
late 1990s, evidence from animal preclinical studies suggested
a major role of food bioactive compounds in the reduction of
colorectal cancer risk (Lipkin et al., 1999; Shike, 1999). Particularly, vegetables and certain bioactive plant components have
been found to possess protective effects on the development of
colorectal cancer and other types of cancer. In recent years much
research interest has focused on rosemary.
Rosemary (Rosmarinus officinalis) belongs to the labiate
family (Lamiaceae). The active constituents of this plant family
are phenolic diterpenes and triterpenes. These compounds have
high antioxidant effects. The main active compounds of rosemary include caffeic acid, rosmarinic acid (RA), ursolic acid
(UA), carnosic acid (CA), and carnosol (Aruoma et al., 1992;
Frankel et al., 1996; al-Sereiti et al., 1999; Moran et al., 2005;
Costa et al., 2007; Anderson et al., 2008). Approximately 90%
of the total antioxidant activity of rosemary is derived from
carnosol and CA (Aruoma et al., 1992; Moran et al., 2005;
Cheung and Tai, 2007). Rosmarinic acid is one of the caffeic

946

Downloaded by [189.200.34.179] at 07:28 19 January 2014

ROSEMARY AND CANCER PREVENTION

derivatives which obtains one of its phenolic rings from phenylalanine via caffeic acid, and the other ring from tyrosine via
3,4-dihydroxyphenyllactic acid. Ursolic acid is a pentacyclic
triterpenoid with anti-cancer and anti-inflammatory properties
(al-Sereiti et al., 1999; Costa et al., 2007; Anderson et al., 2008).
In many parts of the world, rosemary is a common household
plant that is used for many purposes including food flavoring,
drink, and beverages, as well as cosmetic uses. Traditionally,
it was used for relieving renal colic, dysmenorrhea, respiratory disorders, and for stimulating hair growth. This was due to
the antispasmodic properties of rosemary which relaxes trachea
and intestinal smooth muscles, as well as its choleretic activity
(al-Sereiti et al., 1999). Moreover, rosemary has been reported
to be of potential therapeutic benefit in the treatment and/or
prevention of several other health conditions. The reported conditions include asthma, spasmogenic disorders, peptic ulcer, inflammatory diseases, hepatotoxicity, atherosclerosis, ischaemic
heart disease, cataract, and poor sperm motility (al-Sereiti et al.,
1999; Hsieh et al., 2007). Rosemary, in particular its phenolic
constituents, has also been found to exert protective effects on
colonic cancer and other types of cancer through several mechanisms. The reported mechanisms are summarized and discussed
briefly in the section titled Results and Discussion. The anticancer evidence was found to be rather subjective and mainly
from animal and cell culture studies. It is worth mentioning that
most of the reported benefits are not from human randomized
controlled trials.
The aim of the current study was to systematically review
and evaluate the evidence derived from all types of studies published over the last 15 years that examined the protective effects
of rosemary and its active constituents on colorectal cancer as
well as other types of cancer. The present work will summarize
the different molecular targets for cancer prevention by rosemary and provide a critical analysis of the evidence base for the
potential protective effects of rosemary.

METHODS
Studies and Data Sources
This critical review evaluated and summarized scientific evidence derived from all studies published in the last 15 years that
compared rosemary and/or rosemary constituents to a control.
The Medline and Pubmed databases were searched for studies
published in English, from May 1995 to March 2010. These
studies examined the effects of rosemary, and rosemary constituents on colorectal cancer and other types of cancer. Primary
preclinical animal studies cited in the primary-search studies
published any year were also collected and added to the review.
No clinical human studies were identified.
The search terms used were: rosemary, Rosmarinus officinalis, labiate vegetables, caffeic acid, rosmarinic acid, ursolic
acid, carnosic acid, carnosol, colorectal, colonic, rectal, cancer,

947

cancers, prevention, chemoprevention, human, animal, randomized controlled trials, controlled clinical trial, random allocation,
clinical trials, case control, cohort, in vivo, and in vitro studies.
The references from the published studies and journal references
not available electronically were sourced manually.

RESULTS AND DISCUSSION

A total of 36 studies were reviewed, of which 8 animal and
28 in vitro studies were identified. No symptoms of rosemary
toxicity have been reported in the literature. No published human
clinical studies on rosemary and cancers were identified.
Abbreviations used in the text are defined in Tables 1 and 2.
Descriptions of the animal studies are summarized in Table 1.
Different types of diet were employed across the animal studies.
It is important to assess the background diet as this can influence outcomes considerably. It has been reported that many of
the anticancer effects disappeared when the test animals were
switched from the old American Institute of Nutrition (AIN)
sucrose-based diet to a standard starch-based diet (Poulsen et
al., 2001). In the current review, of the 8 animal studies, only one
study used the AIN-76A diet (Moran et al., 2005). This study
demonstrated that carnosol decreased intestinal tumor multiplicity by 46% (P < 0.05). It would be of interest to test whether the
reported anticancer effect will still be observed when a starchbased diet is used.
The diet composition for the other 7 studies varied greatly.
However, all the different diets were mainly starch-based.
One study used a standard food pellet diet (Anderson et al.,
2008); three studies used powdered diets consisting of sucrose
(25%), corn starch (37%), and other supplements (Singletary
and Nelshoppen 1991; Singletary et al., 1996; Singletary and
Rokusek, 1997); one study used a standard diet containing corn
and/or coconut oil (20%) (Amagase et al., 1996); one study used
a standard powdered diet (Kitano et al., 2000); and one study
used a commercial diet (Purina Lab Chow 5001, Ralston-Purina
Co., St. Louis, MO) (Huang et al.,1994). Despite the variability
in diet composition, the anticancer effects were observed in all
studies. It would be of interest to test whether starch and its
constituents, such as amylose and amylopectin, may augment
the anticancer effects of rosemary.
In terms of the bioactives being studied, rosemary extract
was investigated in 6 of the 8 animal studies, carnosol in
3, and UA in 2 studies. The anticancer effects of rosemary
extract were found to be statistically significant in 5 of the
6 identified studies. In the remaining study, the anticancer
effects were observed, but did not reach significance at P <
0.05. In this study (Kitano et al., 2000), a standard powdered
diet was used, whereas other studies employed a standard
diet, that was supplemented with corn and/or coconut oil
(Singletary and Nelshoppen et al., 1991; Singletary et al.,
1996; Singletary and Rokusek, 1997; Amagase et al., 1996).
Whether oil supplements may impact the outcomes in terms
of the anticancer effects of rosemary will require further

948

Rosemary extract
DMBA-induced mammary
adduct N = 20, 32 day old
SD rats

Singletary and
Nelshoppen, 1991

0.5-1%

Reduced incidence of ACF containing 3 crypts in cancer initiation phase

(P < 0.05) Increased colonic SMase activity
Decreased colonic adenomas number (per mouse):
P = 0.021
Treated: 18.0 12.7
Control: 33.1 14.1
Restored enterocyte membrane E-cadherin and -catenin
Decreased DEN-induced GST-P positive (number; area foci/cm2 liver):
P > 0.05
Treated: 9.9 5.0; 1.4 1.0
Control: 13.7 6.0; 2.0 1.4
Induced liver and stomach (GST; OR) carcinogen metabolising enzymes
(nmol/min/mg):
P < 0.05
Treated: 3940 876, 2860 544; 38 6, 442 91
Control: 2580 254, 1948 35; 20 2, 293 61
Decreased (palpable; adenocarcinomas) tumor number (per rat):
P < 0.001
Treated: 1.6 0.3, 1.4 0.4; 2.0 0.5, 1.6 0.4
Control: 5.5 0.7; 5.7 0.8 (carnosol 200, 100 mg)
Treated: 1.2 0.3; 1.5 0.3 (rosemary)
Control: 5.5 0.7; 5.7 0.8
Decreased adduct formation (nmol DMBA/nmol DNA):
P < 0.05
Treated: 149.7 9.6, 142.2 7.2
Control: 245.1 28.8, 197.4 17.2 (rosemary)
Treated: 147.8 7.3; 158.3 31.0
Control: 264.7 16.3 (carnosol; ip rosemary)
Reduced DNA adduct formation:
P < 0.05
Total: 76%; 66% (1%; 0.5% in 20% corn oil)
Anti-dG;dA; syn-dA; dG: > 76%
Total: 45%; 34% (1% in 10%; 5% corn oil)
: 40% (0.5% in 5%, 15% coconut oil)
Reduced tumor number (per mouse):
Total: 99; 68; 40% (rosemary, 3.6; 1.2; 0.4 mg)
: 78; 63; 38% (carnosol, 10; 3; 1 mmol)
: 6145% (UA)
Inhibited TPA-induced ornithine decarboxylase, inflammation,
hyperplasia, and tumor promotion
Reduced mammary tumor: 47% (P < 0.05)
Reduced DNA adduct formation (nmol DMBA/mol DNA):
P < 0.05
Total: 97.6 6.3; 80.0 10.5 (1; 0.5%)
Control: 167.8 14.8
Adduct4; 5; 6; 7: 37.2 4.6, 26.0 5.4; 2.7 0.4,
1.9 0.6; 9.9 0.8, 8.5 1.7;
12.3 0.6, 9.8 1.3 (1, 0.5%)
Control: 74.7 8.6; 5.4 0.9; 18.9 2.0; 21.9
2.2

Results

ANOVA and Fishers

test

Students t-test

ANOVA and Fishers

test

ANOVA and Fishers

test

ANOVA and Fishers

test

Students t-test:
in-signif.

Students test

Students t-test

Analysis

ACF, aberrant crypt foci; Alk-Smase, alkaline Sphingomyelinase; AOM, azoxymethane; CA, Carnosic acid; DEN, diethylnitrosamine; DMBA, dimethylbenz(a)anthracence; GST, glutathione S transferase;
OR, oxygen reductase; SD, Sprague Dawley, RA, Rosmarinic acid; UA, ursolic acid.

B(a)P//TPA- or
DMBA/TPA-induced skin
tumors in mouse

Huang et al., 1994

1.23.6mg
110 mmol
0.12 mmol
Topical

0.51%

Rosemary extract
DMBA-induced mammary
adduct N = 4-5, 36 day old
SD rats

Amagase et al.,1996

Rosemary extract
Carnosol UA

0.5%
100200
200 (Ip, mg/kg)

Rosemary extract
DMBA-induced mammary
Carnosol
adduct N = 5-8, 36 day SD
rats

Singletary et al., 1996

Kitano et al., 2000

10 mg/kg body wt
once daily for 5
day
0.6% by body wt

0.11% in diet for 32

week
1% in diet for 10
week

Dose

Rosemary extract
DEN-induced
hepato-carcinogenesis N =
15, 7 week old F344 rats
Rosemary extract
Singletary and Rokusek, Induction carcinogen
1997
metabolising enzymes, N
= 12, 7 week A/J mice

Moran et al., 2005

Rosemary
constituents

AOM-induced colonic cancer UA

in SD rats
Colonic adenomas N = 10, 5 Carnosol
week old Min/+ mice

Cancer type

Characteristics of included animal preclinical studies

Anderson et al., 2008

Study ID

Table 1

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949

AFB1-induced HepG2 cell

damages

Zunino and Storms,

2009

Rosemay extract

CA Rosemary extract

Myeloid leukemia tumor

bearing Balb/c mice

Sancheti and Goyal,

2006a; 2006b
Sharabani et al, 2006

Carnosol CA

DMBA- induced mouse skin

cancer

Human colonic adenoma

Caco-2 cells

Rosemary

CA

UA

Visanji et al., 2006

Cheung and Tai, 2007

Costa et al., 2007

Human cancer HL60, K562,

MCF7 and MDA-MB-468
cells

IL-1/TNF--induced rat C6
glioma

Russo et al., 2009

Huang et al., 2009

CA Carnosol RA UA

Rosemary constituents

Human melanoma M14; A375 Rosemary extract

cells
ALL (acute lyphoblastic
Carnosol
leukemia)

KB C2 - human
multidrug-resistant cervical
carcinoma cells

Cancer type

Characteristics of included in vitro preclinical studies

Nabekura et al., 2010

Study ID

Table 2

10 M 1%

5001000 mg/kg body

wt

150 M

1/20001/150 dilution

1030 M

20 M

18-27 M

1080 M

50 M

Dose

Analysis

Inhibited P-gp mediated drug efflux (daunorubicin; rhodamine; calcein ANOVA and Dunnetts
test or Students
accumulation ratio):
t-test
P < 0.01
Carsonic acid: 2.67 0.13; 2.50 0.50; 1.12 0.03
Carnosol: 2.39 0.24; 1.40 0.30; 1.30 0.01
RA: 0.9 0.09; no data; 1.12 0.02
UA: 2.33 0.12; 2.20 0.20; 1.07 0.06
Reduced the growth of both cancer cells (P < 0.001) and DNA damage, Students t-test
and induced apoptotic cell death
Students t-test
Delayed chemotherapy-induced DNA damage and blocked apoptotic
terminal events, and reduced% cell death compared to chemotherapy
alone
Reduced IL-1 or TNF--induced cell invasion and eliminated MMP-9 Students t-test
activity/expression (P < 0.01), and inhibited the interaction of
ZIP/p62 and PKC-
Students t-test
Reduced AFB1-induced cancer damage:
ROS level: P < 0.001
Treated: 173; 146% (30; 20 M)
Cell death: P < 0.001; 0.01; 0.05 : 63; 26; 16% (30; 20;
10 M)
8-OH- deoxyguanine conc: P < 0.001 :57% (30 M)
Control: 0%
ANOVA and NewmaAnti-proliferative IC50 : 1/700; 1/400; 1/150; 1/500 dilution
Keuls test
(HL60; K562; MCF; MDA-MB-468) Inhibited
HL60 differentiation:
P < 0.001
Treated: 9.5 2.2%
Control: 2.8 0.8%
Reduced LPS-activated NO production:
P < 0.01; 0.001
Treated: 19.2 2.2; 7.7 1.2 M (1/1000;
1/2000)
Control: 32.6 2.3 M
Reduced IL-1 (P < 0.01) and COX-2 (P < 0.05) mRNA expression Students t-test
Inhibited cell growth IC50: 23 M
Increased cell doubling time:
P < 0.0001 Treated: 140; 120 h (carnosol; CA)
Control: 29 h
Increased G2 /M cell population:
P < 0.01
Treated: > 35; 25% (carnosol; CA)
Control: < 20%
Decreased tumor incidence, burden, yield, average weight and diameter, Students t-test
and prolonged tumor occurrence latency period Reduced lipid
peroxidation (P < 0.05) and depleted levels of glutathione restored
Potentiated 1,25D-induced WEHI-3B D(-) leukemia cell differentiation Students t-test
inhibited cell growth, and induced G0/G1 cell cycle arrest (CA)
Delayed 1,25D-induced tumor appearance and reduced tumor size (P
< 0.05, rosemary extract)
(Continued on next page)

Results

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950

Rat colonic cancer

B16/F10 mouse melanoma

cells

Rat DOX-induced
cardio-myocyte oxidative
stress and heart membrane
lipid peroxidation

Human colon cancer HT-29

cells

Leukemia HL60-G cells

Mouse LPS-induced
macrophage RAW 264.7
cell damages

Rat tBH- induced

erythrocyte/liver/
mitochondria damages

Huang et al., 2005

Chlopskova et al., 2004

Anderson et al., 2003

Danilenko et al., 2003

Lin et al., 2003; 2002

Psotova et al., 2003

Cancer type

10 M
0.0120 M

0.1100 M

CA

Carnosol CA UA RA

RA

2080 M

0200 M

RA

UA

1.2510 M

31.25 mM

Dose

Carnosol

UA

Rosemary constituents

Characteristics of included in vitro preclinical studies (Continued.)

Anderson et al., 2006

Study ID

Table 2

Increased colonic cancer inhibiting marker (alk-SMase; TC-induced

alk-SMase) activity:
P < 0.01; 0.001
Treated: 350; 220%
Control: 100%
Inhibited cell growth (colony number):
P < 0.05
Treated: 7; 80; 85% (5.0; 2.5; 1.25 M)
Control: 100%
Inhibited cell invasion:
P < 0.001; 0.05; 0.05 Treated: 20; 40; 81% (10.0; 5.0;
2.5 M)
Control: 100%
Reduced MMP-9 expression IC50: 5 M
Inhibited AKT, P38, JNK, ERK1/2 phosphorylation, and NFk -B and
c- Jun activation
Oxidative stress cardiomyocyte protection marker (ASAT; LDH; ATP;
troponin 1) total%:
P < 0.05 Treared: 105; 95, 105; 90, 60; 110, 105 (200;
100 M)
Marker ATP/ADP: 8.156 0.19
Control: 3.209 0.312
Inhibited heart membrane lipid peroxidation IC50 :
12.67 0.87; 8.17 0.76 M (microsomes; mitochondria)
Inhibited cell proliferation: 30; 35; 25% (80; 40; 20 M, P < 0.05)
Increased histone-DNA formation, caspase (3; 8; 9), and alk-Smase
activity: 5; 3.5; 3; 1.5; 1.4; 1.87 fold
(P < 0.01; 0.001; 0.01; 0.001; 0.05)
Decreased % high ROS containing cells and mean ROS index:
P < 0.05
Treated: 89.3 10.4; 71.9 8.1%
Control: 100 3.2; 104.4 10.8%
Free radical scavenge IC50 : 0.59; 0.6; 0.49 M (carnosol; CA; RA)
Carnosol:
Inhibited LPS-induced NO production: > 50% (P < 0.01)
IC50 : 9.4 M
Inhibited iNOS protein; mRNA; LPS-induced NF--k B DNA binding:
Treated: 0.2, 0.4, 0.7, 0.8; 0.2, 0.4, 0.5, 0.7; 0.2, 0.4, 0.9, 1 fold
(20, 10, 5, 2.5 M)
Control: 1 fold
Free radical scavenge IC50 and redox potential:
P < 0.01
Treated: 4.75 0.25 M; 0.10 V
Control: 66.30 3.70 M; 0.16 V Inhibited
tBH-induced erythrocyte haemolysis:
P < 0.01
Treated: 37.1 0.2; 91.4 9.0; 91.7 9.0% (0.1;
0.25; 0.50 mM)
Control: 0.4 0%
tBH-induced mitochondria lipoperoxidation IC50 : 0.09 0.01Mm

Results

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Students t-test

ANOVA and Dunnetts

test

Students t-test

Students t-test

Students t-test

Students t-test

Wilcoxon test

Analysis

951

Dorrie et al., 2001

Slamenova et al., 2002

Lymphoblastic leukaemia
SEM RS4/11, MV4/11,
REH, Nalm-6 cells

Human UVA-induced skin

GT-F fibroblasts
Human colon cancer Caco-2
cells

Offord et al., 2002

Carnosol

Rosemary extract

CA

Carnosol

Human epithelial 184B5/HER

cells

Subbaramaiah et al.,
2002

Rosemary extract

Carnosol

n/a

Persson et al., 2003

Rosemary extract

Sotelo-Felix et al., 2002; Rat CCl4 /Cl4 -induced liver

2002
damage

n/a

Leal et al., 2003

1827 M

Students t-test

(Continue on next page)

Students t-test
Induced apoptosis: 4070%
Decreased Bcl-2 protein expression (in SEM; RS411; MV4;11 REH;
Nalm-6):
P < 0.05
Treated: 53.1 8.8; 33.9 8.0; 43.2 3.1; 42.7 2.2; 50.2
7.3%
Control: 100%
(Continued on next page)

% decreased (H2 O2 -induced; FPG- oxidized purines/H2 O2 -induced;

FPG- oxidized purines/visible light-excited MB) DNA break :
P < 0.01; 0.001; 0.01
Treated: 6972; 1220/55-60; 2839/6970%
Control: 7985; 3540/8290; 4041/79-81%

0.3 g/mL

Students t-test

ANOVA and Tukey

test

n/a

n/a

n/a

3.0 0.5 4.7 0.5% Antioxidant activity: 4154% oxidation inhibition

dry weight
3.5 g/50 ml
Inhibited heterocyclic amine formation in MelQx; PhIP; Norharman;
Harman meat:
Treated: 7580; 4050; 4555; 8085%
Control: 100%
65 mg/kg body wt
Reduced liver MDA content and plasma ALT: 69; 50% Normalised
plasma bilirubin; prevented the fall of liver glycogen (P < 0.05);
distorted liver parenchyma; increased liver/plasma carcinogen
metabolising enzyme GST; decreased inflammation, necrosis, and
vacuolation
2060 M
Inhibited PMA-induced PGE2 synthesis (pg/g protein):
P < 0.01
Treated: 740 110; 640 90; 390 100 (20; 40;
60 M)
Control: 950 140
Suppressed PMA/EGF-induced COX-2 expression
0.33M
Inhibited UVA-induced MMP-1 mRNA expression: 90%

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952

Human neuronal IMR-32 cells

Human PMA-induced
mammary epithelial
184B5/HER cell damages

Human AFB1 - induced

bronchial B./liver epithelial
T.CMV cell damages

Human B[a]P-induced
bronchial epithelial
BEAS-2B cell damages

Fiander and Schneider,

2000

Subbaramaiah et al.,
2000

Offord et al., 1997

Offord et al., 1995

5 g/mL

0.66 g/mL

Rosemary extract
Carnosol CA

2.5-30 M

UA

Rosemary extract

10 M

2.510 M

Carnosol CA

CA

Inhibited cell proliferation IC50 : 67 M

Induced growth arrest and cell cycle block in the G1 phase; increased
cyclin-dependent kinase inhibitors p21WAFI/p27Kipl
Increased GST activity and cell viability inhibition IC50 :
P < 0.05
Treated: 467 37, 231 55.8; 258 40, 251 149% (carnosol;
CA)
Control: 100%
Inhibited PMA-induced PGE2 synthesis (pg/g):
P < 0.001 Treated: 200; 400; 700; 700; 850 (30; 15; 10; 5; 2.5 M)
Control: 1400 Inhibited PMA-induced COX-2 expression/promoter
activity; protein
kinase C/ERK1/2/ c-Jun/p38/ AP-1 activation
Inhibited AFB1 -induced DNA adduct formation in 1A2;3A4 expressing
B and T.CMV cells: 85; 67, 30; 48%
Inhibited carcinogen activating enzyme activity in 1A2;3A4
expressing B and T.CMV cells:88; 83; 62; 93%
Inhibited B[a]P-induced DNA adduct formation: 70-80%
Inhibited CYP1A1 activity: 90; 75; 50% (rosemary; carnosol; CA)
Inhibited B[a]P-induced CYP1A1 mRNA: 50%; induced GST and
QR mRNA: 34 fold (carnosol, CA)

n/a

n/a

Students t-test

Students t-test

n/a

AFB1, aflatoxin B1; alk-SMasea, alkaline Sphingomyelinase; ALT, alanine aminotransferase; AP-1, activator protein-1; ASAT, aspartate aminotransferase; ATP/ADP, adenosine tri/diphosphate; B[a]P,
benzo(a)pyrene; CA, Carnosic acid; COX-2, cyclooxygenase-2; DMBA, dimethylbenz(a)anthracence; DOX, doxorubicin; EGF, epidermal growth factor; ERK1/2, extracellular signal-regulated kinase1/2;
FPG, formamidopyrimidine glycosylase; GST, glutathione S transferase; IL-1, Interleukin-1; iNOS, inducible nitric oxide synthase; JNK, jun N-terminal Kinase; LDH, lactate dehydrogenase; LPS,
lipopolysaccharide; MB, methylene blue; MDA, malondialdehyde; MMP-9/-1, metalloproteinase-9/-1; NFK -B, Nuclear Factor-KappaB; NO, nitric oxide; PGE2 , prostaglandin E2; PKC, protein kinase C; PMA,
phorbol 12-myristate 13-acetate; QR, quinone reductase; RA, rosmarinic acid; ROS, reactive oxygen species; tBH, tert-butyl hydroperoxide; TC, taurocholate; TNF-, tumor necrosis factor ; UA, ursolic acid;
ZIP, zeta-interacting protein.

Human myeloid HL-60 and

U937 leukemia cells

Characteristics of included in vitro preclinical studies (Continued.)

Steiner et al., 2001

Table 2

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ROSEMARY AND CANCER PREVENTION

investigations. The Kitano study may have failed to show a

significant effect because of the large inter-individual variability.
Overall, only 2 of the 8 animal studies investigated colorectal
cancer. Carnosol (1% in diet) inhibited colorectal cancer at
initiation, promotion, and progression, in addition to reducing
intestinal tumor multiplicity (Moran et al., 2005). Ursolic acid
(0.11% in diet) reduced the incidence of aberrant crypt foci, one
of the earliest precursors of colorectal adenoma development;
in particular the tumor initiation phase (Anderson et al., 2008).
Other types of cancer investigated include mammary, liver,
and skin cancer. Rosemary extract, carnosol, and UA reduced
the incidence and/or multiplicity of mammary tumor (Singletary
and Nelshoppen 1991; Singletary et al., 1996; Amagase et al.,
1996). This finding is constituent with the data from a number
of human studies (reviewed in Michels et al., 2007). From a
randomized controlled trial (RCT) which involved 34 patients,
foods enriched with bioactive compounds, including rosemary
extract, were found to be a promising adjuvant therapy in advanced breast cancer patients. The mechanism underlying this
effect was considered to be through lowering oxidative-stress
in the patients by a rosemary extract. In this review, rosemary
constituents were also found to decrease the number of skin
tumor (Huang et al., 1994), and rosemary extract reduced hepatic tumor biomarkers such as glutathione S transferase-positive
(GST-P) foci as well as inducing carcinogen metabolising enzymes (Singletary and Rokusek, 1997; Kitano et al., 2000).
Overall, the identified animal studies have provided important
insights into the mechanisms of cancer protection by rosemary
and its constituents.
Descriptions of the cell culture studies are summarized in
Table 2. Only 4 of the 28 studies examined colorectal cancer
cells. Carnosol, CA, and UA inhibited cancer growth and/or
proliferation, in particular, by Caco-2 and HT-29 cells, and the
apoptotic effects were found for UA (Anderson et al., 2003;
Visanji et al., 2006). Rosemary extract also decreased oxidativeinduced DNA damage in Caco-2. The mechanism underlying
this effect was through scavenging free radicals (Slamenova
et al., 2002). Both anti-proliferative and apoptotic effects were
dose and time dependent. It was, however, not feasible to compare the potency of the different bioactive carnosol, CA, and
UA or to establish their minimum effective concentrations due
to the relatively small number of studies available for review.
Several other types of cancer have been investigated in cell
culture studies (Table 2). The results suggest that the anticancer
effects of rosemary are unlikely to be tissue specific. This is because rosemary extract was found to inhibit the growth of cancer
cells and related molecular events in several different organs,
including the breast, liver, stomach, melanoma, and leukemia
cells. Similar findings were obtained for carnosol, CA, and UA.
In addition, the anticancer effects were consistently observed in
both human and different rodent cell lines. This suggested that
the protective effects of rosemary are not species specific. In general, the mechanisms underlying the effects of rosemary against
cancers are either induction of cancer protective markers/factors
(apoptosis, carcinogen metabolising enzymes) or inhibition of

953

tumor promoting events (cell growth and proliferation, DNA

adduct/free radical formation, carcinogen activating enzymes,
lipid peroxidation).
In summary, there is substantial and consistent scientific evidence derived from animal and cell culture studies demonstrating protective effects of rosemary on colorectal cancer and
other types of cancer. Preclinical evidence points to the anticancer properties of rosemary extract, carnosol, CA, UA, and
RA. No studies have reported the anticancer activities of other
constituents such as caffeic acid. The anticancer properties appear to arise through dose-related changes in several molecular
events associated with cancers.

REFERENCES
Al-Sereiti, M.R., Abu-Amer, K.M., and Sen, P. (1999). Pharmacology of rosemary (Rosmarinus officinalis Linn.) and its therapeutic potentials. Indian J
Exp Biol. 37: 124130.
Amagase, H., Sakamoto, K., Segal, E.R., and Milner, J.A. (1996). Dietary rosemary suppresses 7,12-dimethylbenz(a)anthracene binding to rat mammary
cell DNA. J Nutr. 126: 14751480.
ACS (American Cancer Society) (2010). Cancer Facts and Figures 2008,
http://www.cancer.org.
Anderson, D., Cheng, Y., and Duan, R.D. (2008). Ursolic acid inhibits the formation of aberrant crypt foci and affects colonic sphingomyelin hydrolyzing enzymes in azoxymethane-treated rats. J Cancer Res Clin Oncol. 134:101107.
Anderson, D., Nilsson, ., and Duan, R.D. (2006). Ursolic acid and other
pentacyclic triterpenoids stimulate intestinal alkaline sphingomyelinase. Eur
J Lipid Sci Technol. 108: 103108.
Anderson, D., Liu, J.J., Nilsson, A., and Duan, R.D. (2003). Ursolic acid inhibits
proliferation and stimulates apoptosis in HT29 cells following activation of
alkaline sphingomyelinase. Anticancer Res. 23: 33173322.
Aruoma, O.I., Halliwell, B., Aeschbach, R., and Loligers, J. (1992). Antioxidant
and pro-oxidant properties of active rosemary constituents: Carnosol and
carnosic acid. Xenobiotica. 22: 257268.
AIHW (Australian Institute of Health and Welfare) (2010). Australias health
2008, http://www.aihw.gov.au/publications.
CCS (Canadian Cancer Society) - National Cancer Institute of Canada (2010).
Canadian Cancer Statistics 2006, http://www.cancer.ca.
Cheung, S. and Tai, J. (2007). Anti-proliferative and antioxidant properties of
rosemary Rosmarinus officinalis. Oncol Rep. 17: 15251531.
Chlopckova, S., Psotova, J., Miketova, P., Sousek, J., Lichnovsky, V., and
Simanek, V. (2004). Chemoprotective effect of plant phenolics against
anthracycline-induced toxicity on rat cardiomyocytes Part II. Caffeic, chlorogenic and rosmarinic acids. Phytotherapy Res. 18: 408413.
Costa, S., Utan, A., Speroni, E., Cervellati, R., Piva, G., Prandini, A., and
Guerra, M.C. (2007). Carnosic acid from rosemary extracts: A potential
chemoprotective agent against aflatoxin B1. An in vitro study. J Appl Toxicol.
27: 152159.
Danilenko, M., Wang, Q., Wang, X., Levy, J., Sharoni, Y., and Studzinski, G.P.
(2003). Carnosic acid potentiates the antioxidant and proliferation effects
of 1{alpha},25-dihydroxyvitamin D3 in leukemia cells but does not promote
elevation of basal levels of intracellular calcium. Cancer Res. 63: 13251332.
Dorrie, J., Sapala, K., and Zunino, S.J. (2001). Carnosol-induced apoptosis
and downregulation of Bcl-2 in B-lineage leukemia cells. Cancer Lett. 170:
3339.
Fiander, H., and Schneider, H. (2000). Dietary ortho phenols that induce glutathione S-transferase and increase the resistance of cells to hydrogen peroxide are potential cancer chemopreventives that act by two mechanisms: the
alleviation of oxidative stress and the detoxification of mutagenic xenobiotics.
Cancer Lett. 156: 117124.

Downloaded by [189.200.34.179] at 07:28 19 January 2014

954

S. N. T. NGO ET AL.

Frankel, E.N., Huang, S-W., Aeschbach, R., and Prior, E. (1996). Antioxidant
activity of a rosemary extract and its constituents, carnosic acid, carnosol, and
rosmarinic acid, in bulk oil and oil-in-water emulsion. J Agric Food Chem.
44:131135.
Hill, M.J., Morson, B.C., and Bussey, H.J. (1978). Aetiology of adenomacarcinoma sequence in large bowel. Lancet. 1: 245247.
Hsieh, C.L., Peng, C.H., Chyau, C.C., Lin, Y.C., Wang, H.E., and Peng, R.Y.
(2007). Low-density lipoprotein, collagen, and thrombin models reveal that
Rosemarinus officinalis L. exhibits potent antiglycative effects. J Agric Food
Chem. 55: 28842891.
Huang, H.C., Huang, C.Y., Lin-Shiau, S.Y., and Lin, J.K. (2009). Ursolic acid inhibits IL-1beta or TNF-alpha-induced C6 glioma invasion through suppressing the association ZIP/p62 with PKC-zeta and downregulating the MMP-9
expression. Mol Carcinog. 48(6): 517531.
Huang, S.C., Ho, C.T., Lin-Shiau, S.Y., and Lin, J.K. (2005). Carnosol inhibits the invasion of B16/F10 mouse melanoma cells by suppressing
metalloproteinase-9 through down-regulating nuclear factor-kappaB and cJun. Biochem Pharmacol. 69: 221232.
Huang, M.T., Ho, C.T., Wang, Z.Y., Ferraro, T., Lou, Y.R., Stauber, K., Ma,
W., Georgiadis, C., Laskin, J.D., and Conney, A.H. (1994). Inhibition of skin
tumorigenesis by rosemary and its constituents carnosol and ursolic acid.
Cancer Res. 54: 701708.
Leal, P.F., Braga, M.E., Sato, D.N., Carvalho, J.E., Marques, M.O., and Meireles,
M.A. (2003). Functional properties of spice extracts obtained via supercritical
fluid extraction. J Agricul Food Chem. 51: 25202525.
Lipkin, M., Reddy, B., Newmark, H., and Lamprecht, S.A. (1999). Dietary
factors in human colorectal cancer. Annu Rev Nutr. 19: 545586.
Kitano, M., Wanibuchi, H., Kikuzaki, H., Nakatani, N., Imaoka, S., Funae,
Y., Hayashi, S., and Fukushima, S. (2000). Chemopreventive effects of
coumaperine from pepper on the initiation stage of chemical hepatocarcinogenesis in the rat. Jpn J Cancer Res. 91: 67480.
Michels, K.B., Mohllajee, A.P., Roset-Bahmanyar, E., Beehler, G.P., and
Moysich, K.B. (2007). Diet and breast cancer: a review of the prospective
observational studies. Cancer. 109: 2712S2749S.
Moran, A.E., Carothers, A.M., Weyant, M.J., Redston, M., and Bertagnolli,
M.M. (2005). Carnosol Inhibits {beta}-Catenin Tyrosine Phosphorylation
and Prevents Adenoma Formation in the C57BL/6J/Min/ +(Min/+) Mouse.
Cancer Res. 65: 10971104.
Offord, E.A., Gautier, J.C., Avanti, O., Scaletta, C., Runge, F., Kramer, K.,
and Applegate, L.A. (2002). Photoprotective potential of lycopene, [beta]carotene, vitamin E, vitamin C and carnosic acid in UVA-irradiated human
skin fibroblasts. Free Radi Biol Med. 32: 12931303.
Offord, E.A., Mace, K., Avanti, O., and Pfeifer, A.M. (1997). Mechanisms
involved in the chemoprotective effects of rosemary extract studied in human
liver and bronchial cells. Cancer Lett. 114: 275281.
Offord, E.A., Mace, K., Ruffieux, C., Malnoe, A., and Pfeifer, A.M. (1995).
Rosemary components inhibit benzo[a]pyrene-induced genotoxicity in human bronchial cells. Carcinogenesis. 16: 20572062.
Persson, E., Graziani, G., Ferracane, R., Fogliano, V., and Skog, K. (2003).
Influence of antioxidants in virgin olive oil on the formation of heterocyclic
amines in fried beefburgers. Food Chem Toxicol. 41: 15871597.
Potter, J.D., Slattery, M.L., Bostick, R.M., and Gapstur, S.M. (1993). Colon
cancer: a review of the epidemiology. Epidemiol Rev. 15: 499545.
Poulsen, M., Mlck, A.M., Thorup, I., Breinholt, V., and Meyer, O. (2001). The
influence of simple sugars and starch given during pre- or post-initiation on
aberrant crypt foci in rat colon. Cancer Lett. 167: 135143.
Psotova, J., Lasovsky, J., and Vicar, J. (2003). Metal-chelating properties, electrochemical behavior, scavenging and cytoprotective activities of six natural phenolics. Biomedicals Papers of the Medical Faculty of the University
Palacky, Olomouc, Czechoslovakia Republic. 147: 147153.
Russo, A., Lombardo, L., Troncoso, N., Garbarino, J., and Cardile, V. (2009).
Rosmarinus officinalis extract inhibits human melanoma cell growth. Nat
Prod Commun. 4(12): 17071710.

Sancheti, G., and Goyal, P.K. (2006a). Effect of Rosmarinus officinalis in modulating 7,12-dimethylbenz(a)anthracene induced skin tumorigenesis in mice.
Phytother Res. 20: 981986.
Sancheti, G., and Goyal, P. (2006b). Modulatory influence of Rosemarinus
officinalis on DMBA-induced mouse skin tumorigenesis. Asian Pac J Cancer
Prev. 7: 331335.
Sharabani, H., Izumchenko, E., Wang, Q., Kreinin, R., Steiner, M., Barvish,
Z., Kafka, M., Sharoni, Y., Levy, J., Uskokovic, M., Studzinski, G.P., and
Danilenko, M. (2006). Cooperative antitumor effects of vitamin D3 derivatives and rosemary preparations in a mouse model of myeloid leukemia. Int
J Cancer. 118: 30123021.
Shike, M. (1999). Diet and lifestyle in the prevention of colorectal cancer: an
overview. Am J Med. 106: 11S15S.
Singletary, K.W., and Rokusek, J.T. (1997). Tissue-specific enhancement of
xenobiotic detoxification enzymes in mice by dietary rosemary extract. Plant
Foods Hum Nutr. 50: 4753.
Singletary, K., MacDonald, C., and Wallig, M. (1996). Inhibition by rosemary
and carnosol of 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumorigenesis and in vivo DMBA-DNA adduct formation. Cancer Lett.
104: 4348.
Singletary, K.W. and Nelshoppen, J.M. (1991). Inhibition of 7,12 dimethylbenz[a]anthracene (DMBA)-induced mammary tumorigenesis and of in vivo
formation of mammary DMBA-DNA adducts by rosemary extract. Cancer
Lett. 60: 169175.
Slamenova, D., Kuboskova, K., Horvathova, E., and Robichova, S. (2002).
Rosemary-stimulated reduction of DNA strand breaks and FPG-sensitive
sites in mammalian cells treated with H2 O2 or visible light-excited Methylene
Blue. Cancer Lett. 177: 145153.
Sotelo-Felix, J., Martinez-Fong, D., and Muriel De la Torre, P. (2002). Protective effect of carnosol on CCl(4)-induced acute liver damage in rats. Eur J
Gastroenterol Hepatol. 14: 10011006.
Sotelo-Felix, J., Martinez-Fong, D., Muriel, P., Santillan, R.L., Castillo,
D., and Yahuaca, P. (2002). Evaluation of the effectiveness of Rosmarinus officinalis (Lamiaceae) in the alleviation of carbon tetrachlorideinduced acute hepatotoxicity in the rat. J Ethnopharmacol. 81:145
154.
Steiner, M., Priel, I., Giat, J., Levy, J., Sharoni, Y., and Danilenko, M. (2001).
Carnosic acid inhibits proliferation and augments differentiation of human
leukemic cells induced by 1,25-dihydroxyvitamin D3 and retinoic acid. Nutr
Cancer. 41: 13544.
Stryker, S.J., Wolff, B.G., Culp, C.E, Libbe, S.D., Ilstrup, D.M., and MacCarty,
R.L. (1987). Natural history of untreated colonic polyps. Gastroenterology.
93: 10091013.
Subbaramaiah, K., Cole, P.A., and Dannenberg, A.J. (2002). Retinoids
and carnosol suppress cyclooxygenase-2 transcription by CREB-binding
protein/p300-dependent and -independent mechanisms. Cancer Res. 62:
25222530.
Subbaramaiah, K., Michaluart, P., Sporn, M.B., and Dannenberg, A.J. (2000).
Ursolic acid inhibits cyclooxygenase-2 transcription in human mammary
epithelial cells. Cancer Res. 60: 23992404.
Visanji, J.M., Thompson, D.G., and Padfield, P.J. (2006). Induction of G2/M
phase cell cycle arrest by carnosol and carnosic acid is associated with
alteration of cyclin A and cyclin B1 levels. Cancer Lett. 237: 130
136.
Winawer, S.J., Zauber, A.G., OBrien, M.J., Ho, M.N., Gottlieb, L., Sternberg,
S., Waye, J.D., Bond, J., Schapiro, M., Stewart, E.T., Panish, J., Ackroyd, F.,
Kurtz, R.C., and Shik, M. (1993). Randomized comparison of surveillance
intervals after colonoscopic removal of newly diagnosed adenomatous polyps.
N Engl J Med. 328: 901906.
Zunino, S.J. and Storms, D. (2009). Carnosol delays chemotherapy induced
DNA fragmentation and morphological changes associated with apoptosis in
leukemic cells. Nutr Cancer. 61(1): 9410.