496.full

7/23/2019 496.full

http://slidepdf.com/reader/full/496full 1/8

http://ict.sagepub.com/ Integrative Cancer Therapies

http://ict.sagepub.com/content/12/6/496The online version of this article can be found at:

DOI: 10.1177/1534735413485417 2013 12: 496 originally published online 30 April 2013Integr Cancer Ther

JafarabadiMohammad Mohammadzadeh, Elnaz Faramarzi, Reza Mahdavi, Behnam Nasirimotlagh and Mohammad Asgha

Metalloproteinase Enzymes in Rectal Cancer Patients Undergoing ChemoradiotherapyEffect of Conjugated Linoleic Acid Supplementation on Inflammatory Factors and Matrix

Published by:

http://www.sagepublications.com

can be found at:Integrative Cancer Therapies Additional services and information for

http://ict.sagepub.com/cgi/alertsEmail Alerts:

http://ict.sagepub.com/subscriptionsSubscriptions:

http://www.sagepub.com/journalsReprints.navReprints:

http://www.sagepub.com/journalsPermissions.navPermissions:

http://ict.sagepub.com/content/12/6/496.refs.htmlCitations:

What is This?

- Apr 30, 2013OnlineFirst Version of Record

- May 8, 2013OnlineFirst Version of Record

- Oct 17, 2013Version of Record>>

by Gheorghies Alina on October 30, 2014ict.sagepub.comDownloaded from by Gheorghies Alina on October 30, 2014ict.sagepub.comDownloaded from

http://ict.sagepub.com/


http://ict.sagepub.com/content/12/6/496


http://www.sagepublications.com/


http://ict.sagepub.com/cgi/alerts


http://ict.sagepub.com/subscriptions


http://www.sagepub.com/journalsReprints.nav


http://www.sagepub.com/journalsPermissions.nav


http://ict.sagepub.com/content/12/6/496.refs.html

http://online.sagepub.com/site/sphelp/vorhelp.xhtml


http://ict.sagepub.com/content/early/2013/04/29/1534735413485417.full.pdf


http://ict.sagepub.com/content/12/6/496.full.pdf












http://ict.sagepub.com/content/12/6/496.refs.html








7/23/2019 496.full


Integrative Cancer Therapies

12(6) 496 –502© The Author(s) 2013

Reprints and permissions:sagepub.com/journalsPermissions.nav

DOI: 10.1177/1534735413485417ict.sagepub.com

Article

Introduction

Colorectal cancer is one of the most prevalent cancers in the

world, and it is the third most common cancer in both men

and women.

1

Surgery and preoperative chemoradiotherapy(CRT) or postoperative CRT are the main treatments for

rectal cancer.1 In most oncology departments, preoperative

chemoradiation is now the current standard of care for

locally advanced operable rectal cancer.2

It is now known that exposure to clinically relevant

doses of ionizing radiation stimulates various biological

responses at the cell and tissue levels through the early

activation of cytokine cascades.3 Increases in proinflam-

matory cytokines such as tumor necrosis factor α (TNF-

α),interleukin 1 (IL-1), IL-6, and IL-8 as a result of cancer

or cancer treatments may be responsible for the incidence

of symptoms such as pain, fatigue, distress, and so on.3

Moreover, many of these inflammatory downstream

responses to radiation are harmful to normal tissue, and

they gave a survival advantage to tumor cells.3 Therefore,new adjuvant anti-inflammatory therapeutic approaches

may be a useful tool to downregulate inflammatory signal-

ing pathways that include TNF-α, IL-1, and Cox-2

417 ICTXXX10.1177/1534735413485417IntegrativeCancer TherapiesMohammad-zadeh etal

1Tabriz University of Medical Sciences, Tabriz, Iran

Corresponding Author:

Elnaz Faramarzi, Nutrition Research Center, Tabriz University of

Medical Sciences, Tabrzi 51677, Iran.

Email: [email protected]

Effect of Conjugated Linoleic AcidSupplementation on InflammatoryFactors and Matrix MetalloproteinaseEnzymes in Rectal Cancer PatientsUndergoing Chemoradiotherapy

Mohammad Mohammadzadeh, MD1, Elnaz Faramarzi, PhD candidate of

Nutrition1, Reza Mahdavi, PhD1, Behnam Nasirimotlagh, MD1 and

Mohammad Asghari Jafarabadi, PhD1

Abstract

Objectives. The aims of this study were to determine the effect of conjugated linoleic acid (CLA) supplementation oninflammatory factors and matrix metalloproteinase (MMP) enzymes in rectal cancer patients undergoing chemoradiothetrapy.

Method and Material . In this randomized, double-blind, placebo-controlled pilot study, 34 volunteer patients with rectal

cancer undergoing chemoradiotherapy assigned into the CLA group (n = 16), receiving 3 g CLA/d, and placebo group (n =

18) receiving placebo capsules (sunflower oil) for 6 weeks. The supplementation began 1 week before starting RT (loading

period) and continued every day during treatment. Before and after intervention, serum tumor necrosis factor α (TNF-α),interleukin 1β (IL-1β), IL-6, MMP-2, MMP-9, and high-sensitivity C-reactive protein (hsCRP) were measured by enzyme-

linked immunosorbent assay (ELISA) kits and immunoturbidimetric method, respectively. Independent t tests and paired

t tests were used to compare parameters between and within groups, respectively. Results. In the CLA group, the mean

serum TNF-α, IL-1β, hsCRP, MMP-9, and MMP-2 levels reduced insignificantly. However, significant changes in TNF-α (P = 0.04), hsCRP (P = 0.03), and MMP-9 (P = 0.04) concentrations were observed in the CLA group when compared with

the placebo group. The mean serum IL-6 level remained unchanged in the CLA group but increased remarkably in the

placebo group. Conclusion. According to our results, CLA supplementation improved inflammatory factors, MMP-2, and

MMP-9 as biomarkers of angiogenesis and tumor invasion. It seems that CLA may provide new complementary treatmentby reducing tumor invasion and resistance to cancer treatment in patients with rectal cancer.

Keywords

rectal cancer, chemoradiotherapy, conjugated linoleic acid supplementation, inflammatory factors, matrix metalloproteinase

enzymes1

by Gheorghies Alina on October 30, 2014ict.sagepub.comDownloaded from





7/23/2019 496.full


Mohammad-zadeh et al 497

activities for the prevention of radiotherapy (RT) failure

and tumor recurrence and the enhancement of tumor radio-

sensitivity while reducing tumor vascularization and cell

invasion.4-6

Conjugated linoleic acids (CLAs), which represent a

heterogeneous group of positional and geometric isomers

of linoleic acid are found mainly in foods derived from

ruminant animals (such as dairy products and meats).7

CLA consists of a mixture of geometric and positional iso-

mers (cis or trans double-bond positioning at ([7,9],

[8,10], [9,11], [10,12], or [11,13]). The most abundant iso-

mer in dietary sources is cis-9,trans-11-CLA (more than

75%-90% of total CLA).7 The trans-10,cis-12 is the other

main isomer, which represents 1% to 10% of total CLA

from food sources.8 According to findings from animal

and human studies, CLA has beneficial effects on cancer,

inflammation, body composition, diabetes and cardiovas-

cular disease.9 Nowadays, most research is focused on

CLA because of its anti-inflammatory and anticancer

properties. The findings of in vitro and in vivo studiesindicate that CLA has anti-inflammatory effects, for

example, reducing colonic inflammation10 and modulating

the production of cytokines and prostaglandins.11,12

In

humans, CLA has modest anti-inflammatory effects and

improves immune function.13 The results of another clini-

cal trial showed that 6 g of CLA mixture per day in patients

with Crohn’s disease suppressed the production of proin-

flammatory cytokines.14

Furthermore, results from animal

studies showed that CLA supplementation inhibited colon

cancer incidence, progression, and metastasis.15-17 Also,

several in vitro studies have demonstrated that CLA sup-

plementation inhibited metastasis and immigration of

human colon cancer cell line Colo32018 and SW480 cells17

by different mechanisms. Recently, Grądzka et al19

reported that C9,t11-CLA increased sensitivity of HT-29

cells to X radiation. Also, Cho et al20

found that t10,c12-

CLA isomer inhibited HT-29 cells growth via the induc-

tion of G1 cell cycle arrest. The results of another

study indicated that the t10,c12 isomer has more inhibi-

tory effects than the c9,t11 isomer on colorectal cancer

proliferation.21

Anti-inflammatory properties of CLA have been investi-

gated less in human studies, and to the best of our knowl-

edge, there is no published article about the effect of mixed

CLA supplementation in cancer patients. Therefore, weevaluated the effect of CLA supplementation on serum

inflammatory factors and matrix metalloproteinase (MMP)

enzymes as biomarkers of angiogenesis and tumor invasion,

and serum liver enzyme levels in rectal cancer patients

undergoing CRT.

Methods and Materials

This randomized, double-blind, placebo-controlled pilot

study was approved by the Ethics Committee of Tabriz

University of Medical Sciences and registered as an RCT

study (IRCT:201012041197N9). In all, 34 volunteer

patients with rectal cancer who were referred to the RT cen-

ter of Imama Hospital in Tabriz were recruited. Written

informed consent was obtained from all patients.

Those included were ambulatory rectal cancer patients

in stage II or III (based on TNM [tumor-node–metastasis]

staging), who were slated to receive standard preoperative

CRT treatment. RT treatment was administered in 25 frac-

tions of 1.8-Gy (total dose, 45 Gy) concomitant chemother-

apy with 5-fluorouracil and leucovorin (intravenous bolus

infusion for 5 consecutive days in weeks 1 and 5). Exclusion

criteria were history of any other cancer, RT and chemo-

therapy treatments, underweight (BMI < 18.5 kg/m2), vita-

min and mineral supplementation within the past month,

diabetes, and liver, renal, or endocrine dysfunction.

Patients and those involved in doing the assessment and

chemical analyses were blinded to group assignments.

Patients were assigned to the CLA group (n = 16), receiving

four 1000-mg capsules (providing 3 g CLA) 3 times/d (1capsule at breakfast and dinner and 2 capsules at lunch) or

the placebo group (n = 18) receiving 4 placebo capsules for

6 weeks. Placebo capsules were made up with sunflower oil

and were carefully matched in appearance with that of the

CLA capsules. The CLA capsules (Tonalin, Natural factor,

Canada) contained 2 active isomers 18:2 c9, t11 and 18:2

t10,c12 in a 50/50 ratio. The dose of 3 g/d of CLA was cho-

sen on the basis of previous studies carried out in healthy

individuals or in those with other diseases.13,22 The supple-

mentation began 1 week before starting RT (loading period)

and continued every day during RT.

Patients were monitored weekly for any side effects of

supplementation. Compliance was assessed by a capsule

count every 2 weeks. Patients who consumed less than 90%

of the planned number of capsules were excluded from the

study. At the onset of the study, height was measured using

a mounted tape, with the participants’ arms hanging freely

by their sides, and recorded to the nearest 0.5 cm. After

ensuring that they were barefoot and wore light clothing,

their weight was recorded to the nearest 0.1 kg with a Seca

scale. BMI was calculated by dividing weight (in kilo-

grams) by the square of height (in meters).

Before and after intervention, blood samples were col-

lected after an overnight fast of 12 hours. The serums of

patients were kept at −80°C until biochemical analyses.Serum TNF-α, IL-1β, IL-6, and high-sensitivity C-reactive

protein (hsCRP) were measured by platinum enzyme-linked

immunosorbent assay (ELISA) kits (Bender MedSystem

eBioscience, Vienna, Austria) and immunoturbidimetric

method. Serum MMP-2 and MMP-9 were assessed by

ELISA kits (Boster, China, and Bender MedSystem eBio-

science, Vienna, Austria). Serum liver enzyme levels (alka-

line phosphatase [ALP], alanine transaminase [ALT], and

aspartate transaminase [AST]) were determined by the pho-

tometric method.






7/23/2019 496.full


498 Integrative Cancer Therapies 12(6)

Statistical Analysis

The data were analyzed using Statistical Package for theSocial Sciences (SPSS, version 11.5, Chicago, IL).We used

the independent t test on quantitative parameters and the χ 2

test on qualitative variables (gender and stage of disease) to

determine whether baseline characteristics differed between

the CLA and placebo groups. Because all quantitative param-

eters had normal distributions according to the Kolmogorov-

Smirnov test, data were presented as mean ± standard

deviation. The paired t test was used to compare serum bio-

chemical factors at the end of the study with baseline values.

An independent t test was performed to compare changes

(from preintervention to the end of study) of biochemical fac-

tors between the 2 groups. An analysis of covariance test was

used to adjust the effect of confounding factors (baseline val-

ues of biochemical parameters). A P value of less than 0.05

was considered statistically significant.

Results

In this study, 34 patients were recruited. In the CLA group,

1 patient consumed less than 90% of the planned number of

capsules because of forgetting to take CLA capsules regu-

larly and was excluded from the study. In the placebo group,

one patient was hospitalized and excluded from the study.

Finally, statistical analysis was performed on 32 patients

(CLA group, n = 15; placebo group, n = 17). There were nosignificant differences in baseline characteristics between

the 2 groups, as presented in Table 1. The mean biochemical

factors and comparison of changes between the study

groups before and after intervention are indicated in Tables

2 and 3, respectively. Only the baseline values of MMP-9

were significantly ( P = 0.01) different from those of the

control group (Table 2).

After intervention, the mean serum TNF-α and IL-1β

levels did not change significantly within each group (Table

2). However, significant changes ( P = 0.04) of TNF-α levels

(−1.07 pg/mL) were observed in the CLA group when com-

pared with the placebo group (1.8 pg/mL; Table 3).

The mean serum IL-6 levels remained unchanged in the

supplemented group but increased in the placebo group. At

the end of study, the mean serum hsCRP levels reduced in

the CLA group and increased in the placebo group when

compared with baseline (Table 2). The reduction of hsCRP

levels in the supplemented group was significant ( P = 0.03)

in comparison with that in the placebo group (Table 3).

After intervention, the mean serum MMP-9 reduced

insignificantly in both groups. However, changes in MMP-9

levels were −78.1 ng/mL for the CLA group and −23.4 ng/

mL for the placebo group, which indicated a significant dif-

ference from the placebo group ( P = 0.04; Table3). This find-ing was also significant after adjusting for baseline values of

MMP-9.

CLA supplementation reduced serum MMP-2 levels

(−0.093 ng/mL) insignificantly, whereas they increased in the

placebo group (2.85 ng/mL; Table 3) but not significantly.

No significant alterations were observed in ALT and

AST levels within each group during the intervention (Table

2). The mean serum ALP concentration decreased signifi-

cantly ( P = 0.02) in the supplemented group, whereas it did

not change in the placebo group (Table 2).

Discussion

A variety of cytokines (TNF-α, Il-6, Il-1β) and other factors

such as NF-κB and COX2 are involved in the incidence of

inflammation resulting from cancer and cancer treat-

ments.3,17 It has been reported that inflammation plays an

important role in progression, invasion, metastasis, chemo-

resistance, and radioresistance of colorectal cancer.23,24

Therefore, today, many studies are focused on the develop-

ment of new anti-inflammatory therapeutic approaches

for blocking their synthesis or action,25 especially by

Table 1. Baseline Characteristics of Study Groups.

Supplemented Group (n = 15) Placebo Group (n = 17) P

Age (year) 62.4 ± 15.6 58.05 ± 16.4 0.44a

Gender, n (%) 0.75c

Male 8 (53.38%) 10 (58.8%)

Female 7 (46.7%) 7 (41.2%)

Stage rectal cancer, n (%)b 0.78c

II 6 (40%) 6 (35.3%)

III 9 (60%) 11 (64.7%)

Height (cm) 161.7 ± 8.7 156.5 ± 22.9 0.42a

Weight (kg) 66.1 ± 11.2 64.8 ± 10.1 0.73a

BMI (kg/m2) 25.3 ± 4.2 24.6 ± 3.7 0.61a

aIndependent t test.bBased on TNM staging.cχ2 test.






7/23/2019 496.full



dietary supplementation26

such as CLA. The findings from

several animal studies have documented anti-inflammatory

properties of CLA in gut inflammation27,28 and suppressing

colon carcinogenesis and progression.15,29

In pigs with bac-

terial-induced colitis, CLA supplementation decreased

inflammatory colonic lesion developments and upregulated

colonic peroxisome proliferator–activated receptor (PPAR)

γ expression.27

Also, Evans and colleagues29

reported that

CLA supplementation not only attenuated inflammation– induced colorectal cancer in mice but also decreased disease

activity and suppressed colitis-related adenoma and tumor

formation via activation of PPARγ. Furthermore, findings of

other researchers showed that a diet containing 1% t10,c12

or 9t,t11 in rats reduced the incidence and growth of colon

cancer by inducing apoptosis in colonic mucosa of rats

treated with 1,2-dimethylhydrazine.30,31

In a clinical trial,

Bassaganya-Riera et al14

found that CLA supplementation (6

g/d for 12 weeks) in patients with Crohn’s disease sup-

pressed the ability of peripheral blood T cells to produce

proinflammatory cytokines and decreased disease activity.

They concluded that CLA supplementation represents a new

complementary treatment for gut inflammation. Taking into

account the important role of inflammation in tumor forma-

tion and progression of colorectal cancer and the anti-

inflammatory and anti-carcinogenic properties of CLA, we

assumed that dietary CLA ameliorates inflammatory condi-tions and, therefore, it can prevent disease progression. In

support of this hypothesis, the result of our study showed

that 3 g/d CLA supplementation for 6 weeks in rectal cancer

patients undergoing CRT reduced TNF-α levels significantly

as compared with the placebo group. To the best of our

knowledge, there is no published article about the effect of

CLA supplementation on inflammatory factors and MMP

enzymes in rectal cancer patients undergoing CRT.

Table 3. Comparison of Changes in Biochemical Factors Between the Study Groups.a.

CLA Group (n = 15) Placebo Group (n = 17) P b

TNF-α (pg/mL) −1.07 (−6.64, 4.48) 1.88 (−2.66, 6.42) 0.04

IL-1β (pg/mL) −0.32 (−3.26, 2.60) 1.00 (−3.6, 5.69) 0.60

IL-6 (pg/mL) 0.38 (−3.06, 3.83) 4.3 (−2.57, 11.20) 0.26

hsCRP (mg/L) −0.84 (−2.95, 1.26) 1.41 (−0.069, 2.89) 0.03

MMP-9 (ng/mL) −78.1 (−235.2, 78.88) −23.4 (−224.3, 177.5) 0.04

MMP-2 (ng/mL) −0.093 (−9.29, 9.10) 2.85 (−10.07, 15.78) 0.69

ALT (IU/L) 0.21 (−4.3, 4.8) −1.13 (−5.3, 3.12) 0.60

AST (U/L) −0.46 (−3.71, 2.78) −1.73 (−5.09, 1.63) 0.56

ALP (U/L) −28.3 (−52.31, −4.2) −0.85 (−27.7, 26.05) 0.05

Abbreviations: CLA, conjugated linoleic acid; TNF-α, tumor necrosis factor α; IL, interleukin 1β; hsCRP, high-sensitivity C-reactive protein; MMP,matrix metalloproteinase; ALT, alanine transaminase; AST, aspartate transaminase; ALP, alkaline phosphatase.aANCOVA was used to adjust baseline values of biochemical parameters; mean value (95% confidence interval).bP , comparison between groups by independent t test.

Table 2. The Mean Serum Biochemical Factors Before and After Intervention in Both Supplemented and Placebo Groups.a.

CLA Group (n = 15) Placebo Group (n = 17)

Before After P b Before After P b

TNF-α (pg/mL) 55.6 ± 8.67 54.5 ± 3.06 0.68 55.8 ± 7.2 57.7 ± 4.39 0.30

IL-1β (pg/mL) 45.4 ± 5.18 45.1 ± 4.84 0.81 44.7 ± 5.3 45.7 ± 10.84 0.35

IL-6 (pg/mL) 6.1 ± 3.9 6.5 ± 4.5 0.89 8.03 ± 5.4 12.3 ± 11.8 0.19hsCRP (mg/L) 2.5 ± 2.1 1.7 ± 1.5 0.45 2.4 ± 2.26 3.8 ± 3.2 0.06

MMP-9 (ng/mL) 578.01 ± 237.14c 499.8 ± 297.3 0.31 839.1 ± 302.43 815.7 ± 291.72 0.80

MMP-2 (ng/mL) 21.28 ± 16.17 21.19 ± 4.69 0.98 22.26 ± 18.58 25.11 ± 20.74 0.64

ALT (U/L) 15.1 ± 11.4 15.3 ± 6.0 0.90 14.8 ± 9.04 13.6 ± 9.7 0.57

AST (U/L) 15.6 ± 3.2 15.1 ± 5.34 0.31 17.93 ± 4.25 16.2 ± 6.01 0.28

ALP (U/L) 218.7 ± 55.16 190.4 ± 36.95 0.02 221.2 ± 53.60 220.3 ± 56.29 0.94

Abbreviations: CLA, conjugated linoleic acid; TNF-α, tumor necrosis factor α; IL, interleukin 1β; hsCRP, high-sensitivity C-reactive protein; MMP,matrix metalloproteinase; ALT, alanine transaminase; AST, aspartate transaminase; ALP, alkaline phosphatase.aMean value ± standard deviation.bP , comparison within group by paired t test.cBaseline values were significantly different from that in the placebo group.






7/23/2019 496.full



Therefore, we compared our results with in vitro and animal

studies or human studies, which were conducted in healthy

individuals or patients with other diseases.

The results of the study by Yang and Cook indicated that

the plasma TNF-α level after lipopolysaccharide injection

was suppressed in CLA-fed mice.32 Also, Rahman et al33

demonstrated that CLA mixture supplementation for 10

weeks reduced TNF-α levels in mice. In healthy individu-

als, 3 CLA mixtures per day (50:50 c9,t11 CLA and t10,c12

CLA for 12 weeks) decreased TNF-α concentration.13 In

another clinical trial, CLA (cis-9,trans-11 isomer) supple-

mentation in those with birch pollen allergy decreased the

in vitro production of TNF-α.34 The finding of the present

study is in agreement with results of the above-mentioned

studies.13,32-34

Another result of this study was that IL-1β decreased

slightly in the CLA group while increasing in the placebo

group. The findings of other studies showed that CLA sup-

plementation had no effect on serum IL-1β concentration in

healthy individuals.35,36

Previously, Song et al13

reportedthat CLA mixture supplementation (3 g/d for 12 weeks) in

healthy young individuals reduced serum IL-1β levels

significantly.

In this study, CLA supplementation did not change IL-6

levels in cancer patients. Similarly, the findings of previous

studies showed that CLA mixture supplementation had no

effect on serum IL-6 concentration in healthy individu-

als36,37

and patients with diabetes.22

In contrast to the supplemented group, IL-6 levels

increased remarkably in the placebo group, which is com-

patible with the study by Wang et al, who noted that serum

IL-6 concentration increased in non-small-cell lung cancer

during CRT.38 According to our results, CLA supplementa-

tion may prevent the enhancement of IL-1β and IL-6 con-

centrations in patients with cancer during CRT.

At the end of the study, hsCRP levels were elevated in the

placebo group. In line with the present result, Cengiz et al39

observed significant elevation of CRP levels in cancer

patients undergoing pelvic RT. As compared with the pla-

cebo group, CLA supplementation resulted in significant

reduction of hsCRP concentration in the supplemented

group. In contrast to our result, Smedman et al40 observed

significant increases in hsCRP levels in healthy individuals

supplemented with 4.2 g of CLA mixture per day. On the

other hand, the findings of other studies showed that CLAmixture supplementation (3 g/d and 3.2 g/d) had no effect on

CRP levels among overweight37 and obese individuals.41

It can be assumed that the observed difference between

our results and those of previous studies regarding the effect

of CLA on CRP may be a result of different doses of CLA

supplementation used or the health condition of the indi-

viduals. It has been suggested that the anti-inflammatory

properties of CLA are more profound in inflammatory

conditions.34

In this study, the observed modulatory effects of CLA

supplementation on proinflammatory cytokines and

hsCRP in rectal cancer patients can be ascribed to differ-

ences in the mechanisms which that are proposed by in

vitro and in vivo studies. CLA isomers are reported to be a

ligand for PPARγ,42,43 which has a crucial role in the regu-

lation of inflammation, especially inflammatory bowel

disease and the incidence and metastasis of colorectal can-

cer.44,45 The results of several animal studies have shown

that CLA attenuated gut inflammation via activation of

PPARγ.10,27,29

Also, various CLA isomers activate PPARγ

in RAW 264.7 cells and reduce the production of proin-

flammatory cytokines such as TNF-α, IL-1β, and IL-6.45

Therefore, CLA can improve different inflammatory con-

ditions through activation of PPARγ. In addition, CLA

reduces proinflammatory eicosanoid (PGE2, LTB4)

production46,47and attenuates the NF-κB pathway,48 which

is involved in cytokine gene expression, cell cycle activa-

tion, apoptosis, and carcinogenesis.26

It seems that CLA

may have a beneficial effect on inflammation, which influ-ences initiation, progression, and metastasis of colorectal

tumors49and also tumor resistance to cancer treatments

and side effects of these treatments.

In comparison with the placebo group, noticeable

changes in MMP-9 and MMP-2 levels were observed in the

CLA group. As far as we know, this is the first study evalu-

ating the effect of CLA supplementation on MMP-2 and

MMP-9 levels in rectal cancer patients. The present find-

ings in rectal cancer patients are compatible with those of

previous studies.17,50 Soel et al17 noted that MMP-9 activity

in SW480 human colon cancer cells was significantly sup-

pressed by c9,t11 CLA. The results of another study showed

that MMP-2 and MMP-9 synthesis and activity were inhib-

ited by CLA in rats.50 MMP-2 and MMP-9 are 2 important

members of the MMP family that play vital roles in tumor

invasion and angiogenesis; therefore, inhibition of MMP

synthesis or activity in different ways can suppress tumor

invasion and metastasis.51 However, more studies are

needed to clarify the inhibitory mechanism of CLA on the

synthesis and activity of MMP enzymes.

Taking into account that this is the first study to examine

the effect of CLA supplementation in cancer patients, we

determined serum liver enzymes to assess the safety of

CLA in cancer patients. No significant alterations were

observed in ALT and AST levels in the CLA group duringthe intervention. Moreover, the mean serum ALP concen-

trations decreased significantly in the CLA group. The

mean serum liver enzyme levels were in the normal range

in both the groups.

In conclusion, the results of the present study indicate

that CLA supplementation (50:50 cis-9,trans-11 and trans-

10,cis-12 CLA) improved TNF-α and hsCRP levels as indi-

cators of inflammation and MMP-2 and MMP-9 enzymes

as biomarkers of angiogenesis and tumor invasion.





7/23/2019 496.full



Moreover, CLA supplementation prevented the elevation of

IL-1β and IL-6 concentrations as biomarkers of inflamma-

tion when compared with the placebo group in rectal cancer

patients undergoing CRT.

Based on the results of previous animal studies and

the present study, it seems that CLA may provide a

new complementary treatment by reducing tumor invasion

and resistance to cancer treatment in patients with rectal

cancer. However, further studies with larger sample sizes

are needed to achieve more precise results. Moreover,

it would be worthwhile to study the effect of different

doses of oral CLA supplementation on the production of

inflammatory factors and the immune function in cancer

patients.

Acknowledgment

The authors are grateful for the financial support of the Nutrition

Research Center, Tabriz University of Medical Sciences. The

authors also are deeply indebted to all patients who participated in

this study.

Authors’ Note

This article was written based on a data set for the PhD thesis

(Elnaz Faramarzi) registered in Tabriz University of Medical

Sciences.

Declaration of Conflicting Interests

The authors declared no potential conflicts of interest with respect

to the research, authorship, and/or publication of this article.

Funding

The authors disclosed receipt of the following financial support forthe research, authorship, and/or publication of this article: The

authors are grateful for the financial support of the Nutrition

Research Center, Tabriz University of Medical Sciences.

References

1. American Cancer Society. Cancer Facts and Figures 2012.

Atlanta, GA: American Cancer Society; 2012.

2. Kachnic LA. Adjuvant chemoradiation for localized rec-

tal cancer: current trends and future directions. Gastrointest

Cancer Res. 2007;1S:S64-S72.

3. Deorukhkar A, Krishnan S. Targeting inflammatory path-

ways for tumor radiosensitization. Biochem Pharmacol .

2010;80:1904-1914. 4. Dinarello CA. Anti-inflammatory agents: present and future.

Cell . 2010;140:935-950.

5. Balkwill F, Mantovani A. Cancer and inflammation: impli-

cations for pharmacology and therapeutics. Clin Pharmacol

Ther . 2010;87:401-406.

6. Grivennikov SI, Greten FR, Karin M. Immunity, inflamma-

tion, and cancer. Cell . 2010;140:883-899.

7. Chin SF, Storkson JM, Ha YL, et al. Dietary sources of conju-

gated dienoic isomers of linoleic acid. J Food Compos Anal .

1992;5:185-197.

8. Choi JS, Jung MH, Park HS, et al. Effect of conjugated lin-

oleic acid isomers on insulin resistance and mRNA levels

of genes regulating energy metabolism in high fat-fed rats.

Nutrition. 2004;20:1008-1017.

9. Park Y. Conjugated linoleic acid (CLA): good or bad trans fat.

J Food Compos Anal . 2009;225:S4-S12.

10. Bassaganya-Riera J, Hontecillas R. Dietary conjugated

linoleic acid and n-3 polyunsaturated fatty acids in inflam-matory bowel disease. Curr Opin Clin Nutr Metab Care.

2010;13:569-573.

11. Changhua L, Jindong Y, Defa L, et al. Conjugated linoleic

acid attenuates the production and gene expression of proin-

flammatory cytokines in weaned pigs challenged with lipo-

polysaccharide. J Nutr . 2005;135:239-244.

12. Stachowska E, Dolegowska B, Dziedziejko V, et al.

Prostaglandin E2 (PGE2) and thromboxane A2 (TXA2)

synthesis is regulated by conjugated linoleic acids (CLA) in

human macrophages. J Physiol Pharmacol . 2009;60:77-85.

13. Song HJ, Grant I, Rotondo D, et al. Effect of CLA supplemen-

tation on immune function in young healthy volunteers. Eur J

Clin Nutr . 2005;59:508-517.

14. Bassaganya-Riera J, Hontecillas R, Horne WT, et al.Conjugated linoleic acid modulates immune responses in

patients with mild to moderately active Crohn’s disease. Clin

Nutr . 2012;31:721-727.

15. Shiraishi R, Iwakiri R, Fujise T, et al. Conjugated linoleic acid

suppresses colon carcinogenesis in azoxymethane-pretreated

rats with long-term feeding of diet containing beef tallow. J

Gastroenterology. 2010;45:625-635.

16. Kim KH, Park HS. Dietary supplementation of conjugated

linoleic acid reduces colon tumor incidence in DMH-treated

rats by increasing apoptosis with modulation of biomarkers.

Nutrition. 2003;19:772-777.

17. Soel SM, Choi OS, Bang MH. Influence of conjugated lin-

oleic acid isomers on the metastasis of colon cancer cells in

vitro and in vivo. J Nutr Biochem. 2007;18:650-657.

18. Kuniyasu H, Yoshida K, Sasaki T, et al. Conjugated linoleic

acid inhibits peritoneal metastasis in human gastrointestinal

cancer cells. Int J Cancer . 2006;118:571-576.

19. Grądzka I, Sochanowicz B, Brzóska K, et al. Cis-9,trans-11-

conjugated linoleic acid affects lipid raft composition and

sensitizes human colorectal adenocarcinoma HT-29 cells to

X-radiation. Biochim Biophys Acta. 2013;1830:2223-2242.

20. Cho HJ, Kim EJ, Lim SS, et al. Trans-10,cis-12, not cis-

9,trans-11, conjugated linoleic acid inhibits G1-S progression

in HT-29 human colon cancer cells. J Nutr . 2006;136:893-898.

21. Palombo JD, Ganguly A, Bristrain BR, et al. The antipro-

liferative effects of biological active isomers of conjugated

linoleic acid on human colorectal and prostatic cancer cells.Cancer Lett . 2002;177:163-172.

22. Moloney F, Yeow TP, Mullen A, et al. Conjugated linoleic

acid supplementation, insulin sensitivity, and lipoprotein

metabolism in patients with type 2 diabetes mellitus. Am J

Clin Nutr . 2004;80:887-895.

23. Aggarwal BB, Gehlot P. Inflammation and cancer: how

friendly is the relationship for cancer patients? Curr Opin

Pharmacol . 2009;9:351-369.

24. Terzic J, Grivennikov S, Karin E, Karin M. Inflammation and

colon cancer. Gastroenterology. 2010;138:2101-2114.






7/23/2019 496.full



25. Germano G, Allavena P, Mantovani A. Cytokines as a

key component of cancer-related inflammation. Cytokine.

2008;43:374-379.

26. Aggarwal BB, Vijayalekshimi RV, Sung B. Targeting inflam-

matory pathways for prevention and therapy of cancer: short-

term friend, long-term foe. Clin Cancer Res. 2009;15:425-430.

27. Hontecillas R, Wannemeulher MJ, Zimmerman DR, et al.

Nutritional regulation of porcine bacterial-induced colitis byconjugated linoleic acid. J Nutr . 2002;132:2019-2027.

28. Hontecillas R, Bassaganya-Riera J, Wilson J, et al. CD4+

T-cell responses and distribution at the colonic mucosa dur-

ing Brachyspira hyodysenteriae –induced colitis in pigs.

Immunology. 2005;115:127-135.

29. Evans NP, Misyak SA, Schmelz EM, et al. Conjugated linoleic

acid ameliorates inflammation-induced colorectal cancer in

mice through activation of PPARγ. J Nutr . 2010;140:515-521.

30. Park HS, Ryu JH, Ha YL, et al. Dietary conjugated lin-

oleic acid (CLA) induces apoptosis of colonic mucosa in 1,

2-dimethylhydrazine-treated rats: a possible mechanism of the

anticarcinogenic effect by CLA. Br J Nutr . 2001;86:549-555.

31. Park HS, Chun CS, Kim S, Ha YL, Park JH. Dietary trans-

10, cis-12 and cis-9, trans-11 conjugated linoleic acidsinduce apoptosis in the colonic mucosa of rats treated with

1,2-dimethylhydrazine. J Med Food . 2006;9:22-27.

32. Yang M, Cook ME. Dietary conjugated linoleic acid decreased

cachexia, macrophage tumor necrosis factor-α production,

and modifies splenocyte cytokines production. Exp Biol Med .

2003;228:51-58.

33. Rahman MM, Bhattacharya A, Banu J, et al. Conjugated

linoleic acid protects against age-associated bone loss in

C57BL/6 female mice. J Nutr Biochem. 2007;18:467-474.

34. Turpeinen AM, Ylönen N, von Willebrand E, Basu S, Aro A.

Immunological and metabolic effects of cis-9, trans-11-con-

jugated linoleic acid in subjects with birch pollen allergy. Br

J Nutr . 2008;100:112-119.

35. Kelley DS, Taylor PC, Rudolph IL, et al. Dietary conjugated

linoleic acid did not alter immune status in young healthy

women. Lipids. 2000;35:1065-1071.

36. Albers R, Van Der Wielen RP, Brink EJ, et al. Effects of

cis-9, trans-11 and trans-10, cis-12 conjugated linoleic acid

(CLA) isomers on immune function in healthy men. Eur J

Clin Nutr . 2003;57:595-603.

37. Ramakers JD, Plat J, Sebedio JL, et al. Effects of the individ-

ual isomers cis-9,trans-11 vs. trans-10,cis-12 of conjugated

linoleic acid (CLA) on inflammation parameters in moder-

ately overweight subjects with LDL-phenotype B. Lipids.

2005;40:909-918.

38. Wang XS, Shi Q, Williams LA, et al. Inflammatory cytokines

are associated with the development of symptom burden in

patients with NSCLC undergoing concurrent chemoradiation

therapy. Brain Behav Immun. 2010;24:968-974.

39. Cengiz M, Akbulut S, Atahan IL, et al. Acute phase

response during radiotherapy. Int J Radiat Oncol Biol Phys.

2001;49:1093-1096.

40. Smedman A, Basu S, Jovinge S, et al. Conjugated linoleicacid increased C-reactive protein in human subjects. Br J

Nutr . 2005;94:791-795.

41. Steck SE, Chaleci AM, Miller P, et al. Conjugated linoleic

acid supplementation for twelve weeks increases lean body

mass in obese humans. J Nutr . 2007;137:1188-1193.

42. Mc Carty MF. Activation of PPAR gamma may mediate a

portion of the anti cancer activity of conjugated linoleic acid.

Med Hypotheses. 2000;55:187-188.

43. Martin H. Role of PPAR-gamma in inflammation: prospects

for therapeutic intervention by food components. Mutat Res.

2009;669:1-7.

44. Kuniyasu H. The role of dietary PPARγ ligands for metastasis

in colorectal cancer. PPAR Res. 2008;2008:529720.

45. Yu Y, Correl PH, Vanden Heuvel JP. Conjugated linoleic aciddecreases production of proinflammatory products in macro-

phages evidence for a PPAR gamma-dependent mechanism.

Biochem Biophys Acta. 2002;1581:89-99.

46. Li Y, Watkins BA. Conjugated linoleic acids alter bone

fatty acid composition and reduce ex vivo prostaglandin

E2 biosynthesis in rats fed n-6 or n-3 fatty acids. Lipids.

1998;33:417-425.

47. Sugano M, Tsujita A, Yamasaki M, et al. Conjugated linoleic

acid modulates tissue levels of chemical mediators and immu-

noglobulins in rats. Lipids. 1998;33:521-527.

48. Zulet MA, Marti A, Parra MD, et al. Inflammation and conju-

gated linoleic acid: mechanisms of action and implications for

human health. J Physiol Biochem. 2005;61:483-494.

49. Kelley NS, Hubbard NE, Erickson KL. Conjugated linoleic

acid isomers and cancer. J Nutr . 2007;137:2599-2607.

50. Harris MA, Hansen RA, Vidsudhiphan P, et al. Effects of con-

jugated linoleic acids and docosahexaenoic acid on rat liver

and reproductive tissue fatty acids, prostaglandins and matrix

metalloproteinase production. Prostaglandins Leukot Essent

Fatty Acids. 2001;65:23-29.

51. Sbardella D, Fasciglione GF, Gioia M, et al. Human matrix

metalloproteinases: an ubiquitarian class of enzymes

involved in several pathological processes. Mol Aspects Med .

2012;33:119-208.

Documents

496.full