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Prediction of new targets and mechanisms for

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Cite this: DOI: 10.1039/c9fo01168d

quercetin in the treatment of pancreatic cancer,
colon cancer, and rectal cancer
Bing Pang,a Xiaoguang Xu,a Yao Lu,a Han Jin,a Rongrong Yang,a Chunmei Jiang,a
Dongyan Shao,a Yanlin Liub and Junling Shi *a

Received 3rd June 2019,
Accepted 24th July 2019
DOI: 10.1039/c9fo01168d
rsc.li/food-function
Quercetin has been widely found to exhibit anticancer activity with low toxicity and prevalence in foods.
Quercetin has been reported to inhibit digestive system cancers including pancreatic cancer (PAAD) and
colon cancer (COAD), but rectal cancer (READ) has not been reported. The reported mechanisms and
targets are divergent. In this study, new targets and mechanisms were predicted for the influence of quer-
cetin on PAAD, COAD, and READ using bioinformatics methods. The results showed that quercetin may
target CD36 and reduce the death rate caused by PAAD by enhancing the cell adhesion, mediating the
uptake of fatty acids (FAs), regulating thrombospondin-1, and stimulating the immune response.
Quercetin may lower the death rate from READ by targeting SLCO1B1 and producing enhanced effects
from use of this compound, inhibiting cell growth, and inducing apoptosis in tumor cells. ACADS,
ALDH3B2, UGT2A3, AMH, CDKN2A, FOSL1, CD36, CFL2, CYP3A4, and MAF were identified as targets for
quercetin to reduce the death rate caused by COAD. Glutathione metabolism was mainly involved in the
effect of quercetin on COAD, including the enhancement of the oxidation of fatty acids, the metabolism
of anticancer medications, and the stiffness of cells, and the reduction of chemical carcinogenesis, the
level of anti-Müllerian hormone, the proliferation of cancer cells and transcriptional misregulation, and
mediation of the activity of glutathione transferases. The combined analyses of three databases can be
referred to and used to seek medications and targets that can be applied to other diseases.

Introduction problem. It is the third most prevalent cancer worldwide and

Cancers of the digestive tract (mouth, throat, esophagus,
stomach, small intestine, and colorectum) and accessory
organs ( pancreas, gallbladder, and liver) are major cancers of
the digestive system. Among them, pancreatic cancer (PAAD) is
known as a particularly aggressive form of cancer with low
incidence but a high fatality rate. In the United States and
Europe, the mortality rate of PAAD is approximately 3% of all
cancers.1 In clinical terms, colon and rectal cancers are collec-
tively referred to as colorectal cancer. It is a common malig-
nant tumor with high morbidity in the gastrointestinal tract.2
Colorectal cancer is considered a major public health
a
Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences,
Northwestern Polytechnical University, 127 Youyi West Road, Xi’an,
Shaanxi Province 710072, China. E-mail: sjlshi2004@nwpu.edu.cn,
15735356056@163.com, xuxiaoguang@mail.nwpu.edu.cn, 412549599@qq.com,
346221830@qq.com, 1763735068@qq.com, jiangcm@nwpu.edu.cn,
shaoyan@nwpu.edu.cn; Fax: +86-29-88460541; Tel: +86-29-88460541
b
College of Enology, Northwest A&F University, 28 Xinong Road, Yangling,
Shaanxi Province 712100, China
the fourth most frequent cause of death. The incidence of col-
orectal cancer has been declining in recent years due, in large
part, to the increase of colonoscopy screening. However, the
rate of colorectal cancer is still increasing among adolescent
and young adult patients.3 Although there have been many
technological and pharmaceutical advances in the last two
decades, digestive system cancers are still life threatening. It is
imperative to seek new therapeutic targets and medications.
Many natural compounds have been proved to have poten-
tial to inhibit cancers at cellular and animal levels, such as
lignans, lipopeptides and flavonoids.4–6 Among them, querce-
tin is an attractive compound which can be used as a comp-
lementary therapy or dietary agent to treat cancers. Quercetin,
known as 3,3′,4′,5,7-pentahydroxyflavone, is a natural flavonoid
which widely exists in plant flowers, leaves, and fruits in the
form of glycosides, including rutin, quercetin, and hypericin.
Many herbal medicines, such as locust rice, red dates, rhi-
zomes, dioscoreae, mulberry leaves, ginkgo leaves, panax noto-
ginseng, and gold buckwheat, contain this ingredient, and
locust rice flower contains a high content of quercetin (e.g.,
reaching as high as 4%).7 Recently, some endophytic fungi

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Paper
have also been found to produce quercetin and methyl-
quercetin, expanding the availability of these bioactive com-
pounds in nature.8 Quercetin is widely recognized for its high
antioxidant, anti-antiallergic, anti-inflammatory, and antiviral
activities. Recently, the activities of quercetin have been
explored in anti-proliferative and pro-apoptotic effects on
cancer cells of the breast, lung, prostate, bladder, bone, brain,
blood, head and neck, cervix, skin, eye, thyroid gland, ovary,
kidney, and mesothelium,9–12 as well as digestive system
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cancers of the liver, gastric system, colon, and pancreas.
However, to date, there has been no report on the capability of
quercetin to affect rectal cancer.
At cellular levels, the targets and mechanisms by which
quercetin can affect digestive system cancers have been
reported as mitogen-activated protein kinases (MAPKs), AMPK/
mTOR/p38, TLR4 or NF-κB, JNK/JUN, FAK, AKT, MEK1/2, and
ERK1/2 signaling pathways. Proteins of E-cadherin, NF-κB,
p65, TLR4, MMPs (matrix metal proteinases), TIMPs (tissue
inhibitor of metalloproteinases), EMT markers (E-,N-catenin,
Wnt/β-catenin, and snail), and specificity proteins (Sp1, Sp2,
Sp3, and Sp4) have also been reported regarding the effect of
quercetin on digestive system cancers. However, CD36, one of
the most important factors for cancer progress, has not been
reported with regard to the effect of quercetin.13–18
In addition, most of the currently reported targets and
mechanisms for the effect of quercetin on digestive cancers
were obtained at cellular levels, and are rarely identified at
clinical levels. To predict the targets and mechanisms for quer-
cetin in curing digestive system cancers in human patients, we
jointly analyzed the Drug Bank database, String database, and
TCGA database, and used the Kaplan–Meier survival estimate
to predict the possible molecules related to the prognosis.
Materials and methods
Quercetin–target search
The DrugBank database (https://www.drugbank.ca/) was used
to find direct protein targets (DPTs) of quercetin. It is a unique
bioinformatics and cheminformatics resource that combines
detailed drug data with comprehensive drug target
information.
Search for direct protein targets (DPTs) and establishment of a
protein to protein interaction (PPI) network related to
quercetin effects
The STRING database (http://string-db.org/) was used to find
DPTs that may directly interact with quercetin, and a confi-
dence score of >0.5 was set as the cut-off criterion. This data-
base searches for and predicts interactions between known
proteins. It can be applied to 2031 species, containing
9.6 million proteins and 13.8 million proteins. The PPI
network, which was established to evaluate the interaction
between proteins, includes direct physical interactions and
indirect functional correlation.19
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Function enrichment analysis
The Database for Annotation, Visualization and Integrated
Discovery (DAVID) (https://david.ncifcrf.gov/) was used to
perform function enrichment analysis on genes. It provides a
comprehensive set of functional annotation tools to under-
stand the biological meaning behind a large list of genes.
The Cancer Genome Atlas (TCGA) (https://www.cancer.gov/)
was used to obtain the mRNA expression data and clinical data
about pancreatic cancer, colon adenocarcinoma, and rectum
adenocarcinoma. With oversight provided by the National
Cancer Institute and the National Human Genome Research
Institute, the aim is to use high-throughput genome analysis
techniques to help people better understand cancer and
improve methods for preventing, diagnosing, and treating
cancer.
Differentially expressed gene (DEG) search
edgR was used to analyze the data obtained from the TCGA
database. This package analyzes the different genes in
Bioconductor, which provides tools for the analysis and com-
prehension of high-throughput genomic data. Bioconductor
uses the R statistical programming language, is open source
and open development, and is available as an Amazon
Machine Image and a series of Docker images.
Results
DPTs of quercetin and proteins and interaction with DPTs
In an organism, chemicals, proteins, and genes interact with
one another to produce a physiologic function at the cellular
and organ levels. Based on this theory, we queried DrugBank
using quercetin as an input to find targets that interact directly
with quercetin. This resulted in output DB04216 categorizing
quercetin as antioxidants, flavonoids, BCRP/ABCG2 inhibitors,
benzopyrans, chromones, COMT inhibitors, COMT substrates,
cytochrome P-450 CYP2C8 inhibitors, cytochrome P-450
enzyme inhibitors, OATP1B1/SLCO1B1 inhibitors, p-glyco-
protein/ABCB1 inhibitors, protective agents, and pyrans. A
total of 29 quercetin targets were identified (Table 1).
Expanding the search and analysis using the String database,
a total of 884 target proteins (minimum required interaction
score = 0.5, maximum number of interactors = 50), including
the 29 primary targets, were identified to be related to
quercetin.
To assess the functional features of quercetin-mediated
gene sets, KEGG pathway enrichment analysis was performed
using DAVID. The top KEGG pathways (count ≥ 30, FDR < 0.01)
that link to quercetin DPTs and the DPT-associated genes in
cancer were identified for endometrial cancer (30 genes), pan-
creatic cancer (31 genes), colorectal cancer (35 genes), prostate
cancer (47 genes), and pathways in cancer (88 genes) (Fig. 1).
These results suggest that quercetin may play a role in treating
these cancers. All enriched pathways which were identified
using this approach represent biological areas that show a stat-
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Table 1 Proteins interacting with quercetin

DB-ID Name Target symbol Uniprot ID Target symbol Uniprot ID

DB04216 Quercetin HCK P08631 CYP1B1 Q16678

DB04216 Quercetin PIK3CG P48736 ACTB P60709
DB04216 Quercetin UGT3A1 Q6NUS8 CSNK2B P67870
DB04216 Quercetin ATP5A1 P25705 EIF3F O00303
DB04216 Quercetin ATP5B P06576 CSNK2A1 P68400
DB04216 Quercetin ATP5C1 P36542 HSP90AA1 P07900
DB04216 Quercetin PIM1 P11309 HSPA2 P54652
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DB04216 Quercetin ttgR Q9AIUO RUVBL2 Q9Y230

DB04216 Quercetin HIBCH Q6NVY1 SF3B3 Q15393
DB04216 Quercetin fabz Q5G940 UBA1 P22314
DB04216 Quercetin STK17B Q94768 SHBG P04278
DB04216 Quercetin ESR1 P03372 CBR1 P16152
DB04216 Quercetin ESR2 Q92731 CEBPB P17676
DB04216 Quercetin NQO2 P16083 NR1I2 O75469
DB04216 Quercetin AHR P35869

istically significant association with quercetin gene sets, thus

warranting further investigation.

DEGs in digestive system cancers

First, we downloaded the mRNA expression data and clinical
data related to PAAD (4 normal and 178 cancer), COAD
(41 normal and 480 cancer), and READ (10 normal and 167
cancer) from TCGA. By using the edgR packages, the differen-
tially expressed genes (absolute value of log FC > 2) were identi-
fied by comparing the differences of the expression levels
between the normal and cancer samples. The results demon-
strated a total of 1666 DEGs for COAD with 912 up-regulated
and 754 down-regulated DEGs (Fig. 2a); 299 DEGs were identi-
fied for PAAD with 28 up-regulated (log FC > 2) and 271 down-
regulated (log FC < −2) DEGs (Fig. 2b), and 2235 DEGs for
READ with 1120 up-regulated and 1115 down-regulated DEGs
(Fig. 2c).

Identification of intersection genes

Fig. 1 Top KEGG pathways (count ≥ 30, FDR < 0.01) of DPTs and pro-
teins interact with DPTs. The ordinate is the name of each pathway;
abscissa is the number of genes in pathways; and bubble size indicates
the percentage of genes located in a functional area. DPTs refer to the
direct protein targets of quercetin.
To search for the potential targets of quercetin in treating
cancers, we took into account the intersection in IPQ (interact-
ing proteins with quercetin) and DEGs among the three
cancers tested. According to the results (Fig. 3), CD36 exists in
the four parts. It was known that the protein encoded by CD36
is the fourth major glycoprotein of the platelet surface and

Fig. 2 Identification of differentially expressed genes (DEGs) in COAD (a), PAAD (b), and READ (c). The red dots represent all the up-regulated
genes, the black dots represent the normal genes, and the green dots represent all the down-regulated genes.

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As shown in Fig. 4 and Table 3, in PAAD, 8 genes were

identified as responding to immune response (GO:0006955),
innate immune response (GO:0045087), and some proteins in
the membrane, indicating that quercetin may influence PAAD
via immunological response. For COAD, the shared genes are
related to glutathione transferase activity (GO:0004364), the
glutathione derivative biosynthetic process (GO:1901687), and
the glutathione metabolic process (GO:0006749), indicating
that quercetin may affect COAD by glutathione metabolism. In
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READ, the genes are mainly related to oxidation–reduction

(GO:0055114, GO:0019825, GO:0016491, GO:0016712, and
GO:0016705) and the endoplasmic reticulum membrane
(GO:0005789).

Fig. 3 Intersection situation of IPQ and DEGs. IPQ refers to the proteins
interacting with quercetin.
serves as a receptor for thrombospondin in platelets and
various cell lines. IPQ shares 69 genes with READ, 45 genes
with COAD, and 8 genes with PAAD.
Function enrichment
KEGG function enrichment was performed on all of the
shared genes. According to the results (FDR < 0.01), the genes
were annotated in two pathways of cytochrome
P450 metabolism with a total of 51 counts, including 26 in the
chemical carcinogenesis pathway (hsa05204), 20 in retinol
metabolism (hsa00830), 15 in steroid hormone biosynthesis
(hsa00140.), 12 in drug metabolism-other enzymes (hsa00983),
10 in ascorbate and aldarate metabolism (hsa00053), 10 in
pentose and glucuronate interconversions (hsa00040), and 10
in porphyrin and chlorophyll metabolism (hsa00860)
(Table 2).
PPI network of shared genes in three cancers
We established the network of shared genes in three cancers.
Fig. 5 shows the nodes and edges in the network in different
cancers. In COAD, the PPI network had 45 nodes and 147
edges with an average aggregation coefficient of 6.53, and
enrichment p < 0.001 (Fig. 5a). In PAAD, the PPI network had 6
nodes and 5 edges with an average aggregation coefficient of
1.25 and enrichment p = 0.0162 (Fig. 5b). In READ, the PPI
network had 69 nodes and 376 edges with an average aggrega-
tion coefficient of 10.9 and enrichment p < 0.001 (Fig. 5c).
Kaplan–Meier survival estimate
To find the targets of quercetin in clinical samples, we first
divided the reported cancer samples into a high expression
group and a low expression group according to the mid-value
of the significantly expressed genes. We performed Kaplan–
Meier survival estimation of all shared genes for each cancer
and identified the molecules related to prognosis. Screening at
the level of p ≤ 0.05, 1 gene (CD36, p = 0.02354) was identified
for PAAD, 1 gene for READ (SLCO1B1, p = 0.00905), and

Table 2 KEGG enrichment of common genes

Pathway name Count Genes FDR

hsa00980: metabolism of 26 CYP3A4, CYP2B6, ADH6, ADH1B, GSTM5, UGT1A6, UGT1A8, ADH4, UGT1A4, 3.97 × 10−28
xenobiotics by cytochrome P450 UGT2A3, GSTA1, GSTA2, GSTT2B, CYP1A1, SULT2A1, CYP2C9, ALDH3B2, CYP2E1,
CYP1A2, UGT1A1, UGT1A10, UGT2B17, UGT2B11, CYP2A6, UGT2B4, UGT2B15
hsa00982: drug metabolism – 25 GSTA1, CYP3A4, GSTA2, GSTT2B, CYP2B6, CYP2C9, CYP2C8, ADH1B, ADH6, 2.08 × 10−27
cytochrome P450 ALDH3B2, CYP1A2, CYP2E1, UGT1A1, GSTM5, UGT1A10, UGT1A6, UGT2B17,
UGT1A8, ADH4, UGT1A4, UGT2B11, UGT2B4, CYP2A6, UGT2A3, UGT2B15
hsa05204: chemical 26 CYP3A4, ADH6, ADH1B, GSTM5, UGT1A6, UGT1A8, ADH4, UGT1A4, UGT2A3, 3.91 × 10−27
carcinogenesis GSTA1, GSTA2, GSTT2B, CYP1A1, SULT2A1, CYP2C9, CYP2C8, ALDH3B2, CYP2E1,
CYP1A2, UGT1A1, UGT1A10, UGT2B17, UGT2B11, CYP2A6, UGT2B4, UGT2B15
hsa00830: retinol metabolism 20 CYP3A4, CYP1A1, CYP2B6, CYP2C9, CYP2C8, ADH1B, ADH6, CYP1A2, UGT1A1, 2.09 × 10−19
UGT1A10, UGT1A6, UGT2B17, UGT1A8, ADH4, UGT1A4, UGT2B11, CYP2A6,
UGT2B4, UGT2A3, UGT2B15
hsa00140: steroid hormone 15 CYP3A4, HSD17B2, CYP1A1, CYP1A2, CYP2E1, UGT1A1, UGT1A10, UGT1A6, 3.09 × 10−12
biosynthesis UGT2B17, UGT1A8, UGT1A4, UGT2B11, UGT2B4, UGT2A3, UGT2B15
hsa00983: drug metabolism – 12 CYP3A4, UGT1A10, UGT1A6, UGT2B17, UGT1A8, UGT1A4, UGT2B11, UGT2B4, 3.98 × 10−9
other enzymes CYP2A6, UGT2A3, UGT2B15, UGT1A1
hsa00053: ascorbate and aldarate 10 UGT1A10, UGT1A6, UGT2B17, UGT1A8, UGT1A4, UGT2B11, UGT2B4, UGT2A3, 1.48 × 10−8
metabolism UGT2B15, UGT1A1
hsa00040: pentose and 10 UGT1A10, UGT1A6, UGT2B17, UGT1A8, UGT1A4, UGT2B11, UGT2B4, UGT2A3, 1.15 × 10−7
glucuronate interconversions UGT2B15, UGT1A1
hsa00860: porphyrin and 10 UGT1A10, UGT1A6, UGT2B17, UGT1A8, UGT1A4, UGT2B11, UGT2B4, UGT2A3, 1.21 × 10−6
chlorophyll metabolism UGT2B15, UGT1A1

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Fig. 4 GO function enrichment analysis on the shared genes. GO refers to gene ontology; MF refers to molecular function; BP refers to the biologi-
cal process; and CC refers to the cell component.
10 genes for COAD (ACADS, p = 0.04706; ALDH3B2, p =
0.01829; UGT2A3, p = 0.01827; AMH, p = 0.02271; CDKN2A, p =
0.00739; FOSL1, p = 0.04264; CD36, p = 0.0455; CFL2, p =
0.01355; CYP3A4, p = 0.00486; and MAF, p = 0.02138). These
genes are different from the currently reported targets in cell
experiments.
In PAAD, the CD36 high expression group has a higher sur-
vival rate than the low expression group, indicating that up-
regulation of this gene can suppress the progress of PAAD
(Fig. 6a). In READ, the SLCO1B1 (solute carrier organic anion
transporter family, member 1B1) high expression group has a
higher survival rate than the low expression group. Therefore,
up-regulation of SLCO1B can inhibit READ (Fig. 6b). However,
in COAD, the ACADS, ALDH3B2, UGT2A3, AMH, CDKN2A,
FOSL1, CD36, CFL2, and MAF high expression group shows a
lower survival probability than the low expression group, indi-
cating that up-regulation of these genes may accelerate the pro-
gression of COAD. In contrast, the CYP3A4 high expression
group has a higher survival rate than the low expression group,
indicating that down-regulation of this gene expression can
suppress the progression of COAD (Fig. 6c–l).
Predicted mechanisms for quercetin to cure digestive cancers
From the combined results of the function enrichment ana-
lysis and estimated survival rate-related gene expression, it can
be deduced that quercetin may suppress the progression of
PAAD, COAD, and READ in different ways (Fig. 7). For PAAD,
quercetin may enhance the adhesion of cells, mediate the
uptake of FAs, regulate TSP1 by increasing the expression of
CD36, and stimulate the immune response to prevent cancer.
In READ, quercetin may up-regulate the expression of
SLCO1B1, regulate the polymorphism of SLCO1B1, and affect
the uptake of medications to cure cancer. Quercetin can also
enhance the balance of redox reactions and affect the growth
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and apoptosis of cells in READ. In COAD, quercetin may affect
the progression of COAD in the following ways: impeding the
oxidation of fatty acids via down-regulation of ACADS
expression, enhancing the metabolism of anticancer medi-
cations and reducing chemical carcinogenesis by down-regu-
lation of ALDH3B2 and UGT2A3 expression, improving
CYP3A4 expression, reducing the level of anti-Müllerian
hormone, and lowering the risk of COAD by down-regulating
the expression of AMH, performing anticancer function in the
later stage by up-regulation of CDKN2A, reducing the prolifer-
ation of cancer cells by reducing FOSL1 expression, reducing
the expression of CFL2 to enhance the stiffness of cells, retard-
ing transcriptional misregulation by down-regulating the
expression of MAF, and preventing cancer by mediating the
metabolism of glutathione.
Discussion
Three databases, including the DrugBank database, TCGA,
and STRING, were jointly used in this study. They provided
informative and important sources for predicting the possible
targets and mechanisms with respect to the effect of quercetin
on different cancers. The DrugBank database combines
detailed drug (chemical, pharmacological, and pharma-
ceutical) data with comprehensive drug target (sequence,
structure, and pathway) information. It is a comprehensive,
freely accessible, online database containing information on
drugs and drug targets. TCGA is a landmark cancer genomics
program, molecularly characterizing over 20 000 primary
cancers and matched normal samples spanning 33 cancer
types. Up to now, TCGA has generated over 2.5 petabytes of
genomic, epigenomic, transcriptomic, and proteomic data.
The STRING is a database used for searching and predicting
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Table 3 GO terms and names

Term Name

GO:0001972 Retinoic acid binding

GO:0004024 Alcohol dehydrogenase activity, zinc-dependent
GO:0004364 Glutathione transferase activity
GO:0004497 Monooxygenase activity
GO:0004715 Non-membrane spanning protein tyrosine kinase activity
GO:0005496 Steroid binding
GO:0005506 Iron ion binding
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GO:0005789 Endoplasmic reticulum membrane

GO:0005829 Cytosol
GO:0006069 Ethanol oxidation
GO:0006629 Lipid metabolic process
GO:0006749 Glutathione metabolic process
GO:0006805 Xenobiotic metabolic process
GO:0006955 Immune response
GO:0007169 Transmembrane receptor protein tyrosine kinase
signaling pathway
GO:0008152 Metabolic process
GO:0008202 Steroid metabolic process
GO:0008392 Arachidonic acid epoxygenase activity
GO:0008395 Steroid hydroxylase activity
GO:0009813 Flavonoid biosynthetic process Fig. 5 Established PPI network of the shared genes in three
GO:0015020 Glucuronosyltransferase activity
cancers. a. COAD; b. PAAD; and c. READ.
GO:0016491 Oxidoreductase activity
GO:0016758 Transferase activity, transferring hexosyl groups
GO:0017144 Drug metabolic process
GO:0019373 Epoxygenase P450 pathway
GO:0019825 Oxygen binding
GO:0019899 Enzyme binding
products always have low toxicity compared with manually pro-
GO:0020037 Heme binding duced chemical compounds. They also have wide accessibility,
GO:0030154 Cell differentiation efficacy, and a broad range of activities. As a kind of natural
GO:0031090 Organelle membrane
GO:0031234 Extrinsic component of the cytoplasmic side of the
flavonoid, quercetin plays an important role in protecting
plasma membrane against free radical damage, aging, and inflammation.
GO:0038083 Peptidyl-tyrosine autophosphorylation Although there has been a wide range of biological and cellu-
GO:0042738 Exogenous drug catabolic process
GO:0043306 Positive regulation of mast cell degranulation
lar activities for quercetin in protecting against cancers, this
GO:0045087 Innate immune response study is, to the best of our knowledge, one of the few reports to
GO:0045121 Membrane raft focus for the first time on the capability of quercetin to treat
GO:0045922 Negative regulation of the fatty acid metabolic process
GO:0045939 Negative regulation of the steroid metabolic process
digestive cancers, especially rectal cancer.
GO:0051552 Flavone metabolic process New targets and mechanisms were predicted for the anti-
GO:0052695 Cellular glucuronidation cancer activity of quercetin toward digestive system cancers. It
GO:0052696 Flavonoid glucuronidation
GO:0052697 Xenobiotic glucuronidation
has previously been reported that quercetin performed the
GO:0055114 Oxidation–reduction process anticancer activity by binding to cellular receptors and pro-
GO:0097267 Omega-hydroxylase P450 pathway teins or combining with chemotherapeutic agents.20,21
GO:1901687 Glutathione derivative biosynthetic process
GO:1904224 Negative regulation of glucuronosyltransferase activity
Previous studies have primarily been conducted in vivo at cellu-
GO:2001030 Negative regulation of cellular glucuronidation lar levels, and the results indicated that quercetin could
GO:0016705 Oxidoreductase activity, acting on paired donors, with induce apoptosis, inhibit the synthesis of heat shock proteins,
incorporation or reduction of molecular oxygen
GO:0016712 Oxidoreductase activity, acting on paired donors, with
and inhibit many enzymes, including tyrosine protein kinase,
incorporation or reduction of molecular oxygen, reduced phospholipase A2, phosphodiesterase mitochondrial ATPase,
flavin or flavoprotein as one donor, and incorporation of PI3-kinase, and protein kinase C. It can also activate Ca2+ and
one atom of oxygen
K+ channels and behave as an agonist at estrogen (GPER)
receptors.22,23 It has been reported that, in PAAD, quercetin
can reduce the expression of cellular FLICE-like inhibitory
interactions between known proteins. More research continues protein, activate c-Jun N-terminal kinase (JNK),17 and inhibit
to mine each database. In this study, we conjointly analyzed the EGFR-mediated FAK, AKT, MEK1/2, and ERK1/2 signaling
the data of these databases. pathways.24 It can also activate caspase-3, -8, and -9 and reduce
The study revealed that quercetin might have the potential the mitochondrial membrane potential.15,16 However, in this
to treat digestive cancers with target proteins and mecha- study, the target of quercetin in PAAD was identified as CD36,
nisms. In recent years, natural products such as paclitaxel, being different from all previously reported results. According
resveratrol, and flavonoids have shown considerable potential to the current results, in COAD, quercetin can inhibit the cell
in the fight against diseases, such as digestive disorders, viability of CT26 and MC38 colon cancer cells and induce
cancer insurgence, and immune dysfunction.10 These natural apoptosis via the MAPK pathways. It can also regulate the

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Fig. 6 Kaplan–Meier survival estimate of common genes (P < 0.05). a. Survival curve of CD36 in PAAD; b. survival curve of SLCO1B1 in READ; and
c–l. survival curve of common genes in COAD. Red line refers to the gene high expression group. Green line refers to the gene low expression
group.
expression of E-cadherin, NF-κB, p65, TLR4 (anti-toll-like
receptor), CB1-R (endocannabinoid reporter), and EMT
markers (E-,N-catenin, Wnt/β-catenin, and snail).25 In this
study, ACADS, ALDH3B2, UGT2A3, AMH, CDKN2A, FOSL1,
CD36, CFL2, CYP3A4, and MAF were predicted as the targets
of quercetin to treat COAD. These differences may be due to
tumor cells that were used in previous studies, whereas mainly
clinical samples were used in this study. Besides the known
digestive cancers, it was successfully predicted that READ may
be treated with quercetin via the target of SLCO1B1. These
targets may be diagnosis/prognosis markers and may contrib-
ute to the therapeutic strategies for PAAD, COAD, and READ.
CD36 was initially identified as the target of quercetin to
treat PAAD. CD36 is the fourth major glycoprotein of the plate-
This journal is © The Royal Society of Chemistry 2019
View Article Online
Paper
let surface and serves as a receptor for thrombospondin in
platelets and various cell lines. Thrombospondins may have
important functions as a cell adhesion molecule because they
are widely distributed proteins involved in a variety of adhesive
processes.25–27 CD36 is also known as fatty acid translocase,
which functions as a transmembrane protein and mediates
the uptake of FAs. It has been observed as highly expressed in
breast cancer tissues. Consistent with FA synthesis, FA uptake
and transport are also important target pathways for anti-
cancer therapy. Therefore, the FA channel protein CD36 may
provide a promising therapeutic target.28 CD36 has also been
found to be associated with gastric cancer by regulating throm-
bospondin-1 (TSP1), which is correlated with carcinogenesis
occurring in the cases of intestinal inflammation.29 However,
Food Funct.

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Fig. 7 Predicted mechanisms for quercetin suppressing the progression of PAAD, COAD, and READ. → refers to up-regulation, and ⊥ refers to
down-regulation.
it has not been reported that quercetin can mediate cancers by
regulating CD36. In this study, CD36 was found to be highly
related to the occurrence of PAAD and COAD. In these two
cancers, the CD6 high expression group showed a higher survi-
val rate than the CD6 low expression group. This illustrated
that quercetin may enhance the adhesion of cells, mediate the
uptake of FAs, and regulate TSP1 by up-regulating the
expression of CD36 in these cancers.
SLCO1B1 was identified as the target of quercetin in COAD.
This has not been previously reported. In terms of function,
the protein encoded by this gene is a transmembrane receptor
which mediates the sodium-independent uptake of numerous
endogenous compounds.30 It has been reported that SLCO1B1
acts as a cellular uptake transporter for curcumin and its
major metabolites, and may also be applicable to patients
undergoing breast cancer therapy.31 Quercetin and curcumin
are both plant polyphenols. Therefore, SLCO1B1 may play a
similar role in the mechanisms of the treatment of READ by
quercetin. SLCO1B1 also has been reported to harbor multiple
common polymorphism associated with methotrexate clear-
ance, and methotrexate has been used to treat autoimmune
diseases and malignancies, including acute lymphoblastic leu-
kemia.32 In this study, the SLCO1B1 high expression group
also has a higher survival rate than the SLCO1B1 low expression
group. Currently, more and more studies show multiple drugs
used together being more efficient than a single drug in treat-
ment of cancers. Therefore, it can be deduced that quercetin
could enhance the effect of other chemotherapeutic drugs
when they are used in combination, since it can enhance the
expression of SLCO1B1, regulate the polymorphism of
SLCO1B1, and consequently affect the uptake of drugs to cure
READ. This mechanism might also be suitable for other com-
pounds with similar structures to quercetin, such as curcumin.
For COAD, many proteins related to metabolism were pre-
dicted as the potential targets for quercetin, which had not
been previously reported. In terms of functions, these proteins
are related to cell proliferation, actin filament depolymeriza-
Food Funct.
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Food & Function
tion, transcriptional misregulation, stabilization of the tumor
suppressor protein p53, the mitochondrial fatty acid beta-oxi-
dation pathway, metabolism of xenobiotics by cytochrome
P450, and the mediation of male sexual differentiation. They
are detailed as follows.
(1) FOSL1 (FOS-like antigen 1) enables positive regulation of
cell proliferation.33,37 The FOSL1 high expression group,
showing a lower survival rate, indicated that quercetin may
reduce the proliferation of cancer cells by reducing the
expression of FOSL1.
(2) MAF (v-maf avian musculoaponeurotic fibrosarcoma
oncogene homolog) is related to transcriptional misregulation
in cancer.33 In this study, the MAF high expression group
resulted in a low survival rate, indicating that transcriptional
misregulation may accelerate the progression of cancer.
Quercetin can retard transcriptional misregulation by reducing
the expression of MAF and, thus, cure COAD.
(3) CFL2 (cofilin 2) has been reported to enable positive
regulation of actin filament depolymerization. The actin fila-
ment is associated with the mechanical features of cells. Actin
filament depolymerization can reduce the stiffness of cells.35
In this study, the CFL2 high expression group had a lower sur-
vival rate, suggesting that these cells have a lower degree of
stiffness. It was predicted that quercetin may reduce the
expression of CFL2 to enhance the stiffness of cells and, thus,
cure COAD.
(4) CDKN2A (cyclin-dependent kinase inhibitor 2A) is an
inhibitor of CDK4 kinase. The product of CDKN2A is a stabil-
izer of the tumor suppressor protein p53 as it can interact with
and sequester the E3 ubiquitin-protein ligase MDM2 (a
protein responsible for the degradation of p53). This gene is
frequently mutated or deleted in a variety of tumors, and it is
recognized as an important tumor suppressor gene.36 In this
study, the CDKN2A high expression group had a lower survival
rate at the early stage but a higher survival rate in the later
stage. Quercetin might be able to perform its function at the
later stage by enhancing the expression of CDKN2A.
This journal is © The Royal Society of Chemistry 2019

Food & Function
(5) ACADS (acyl-CoA dehydrogenase, C-2 to C-3 short chain)
catalyzes the initial step of the mitochondrial fatty acid beta-
oxidation pathway. It was also identified as a potential diagno-
sis/prognosis marker in bladder cancer.34 In this study, it was
predicted that quercetin may impede the oxidation of FA by
reducing the ACADS expression and finally lower the carcino-
genesis of COAD.
(6) ALDH3B2 (aldehyde dehydrogenase 3 family, member
B2), UGT2A3 (UDP glucuronosyltransferase 2 family, polypep-
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tide A3), and CYP3A4 (cytochrome P450, family 3, subfamily A,
polypeptide 4) are the three molecules involved in the metab-
olism of xenobiotics by cytochrome P450, drug metabolism–
cytochrome P450, and chemical carcinogenesis pathways.
Cytochrome P450 is involved in the metabolism of various
anticancer agents, such as cyclophosphamide, ifosfamide,
tamoxifen, docetaxel, and paclitaxel. In this study, the
ALDH3B2 and UGT2A3 high expression group showed a low
survival rate, whereas the CYP3A4 high expression group
exhibited a high survival rate. It was predicted that quercetin
may suppress the expression of ALDH3B2 and UGT2A3, up-
regulate the expression of CYP3A4, enhance the metabolism of
anticancer agents, and reduce chemical carcinogenesis in
COAD. These suggest that quercetin can be used as an assist-
ance agent to improve the medications used for treating and
potentially curing cancer.34
(7) AMH (anti-Müllerian hormone) is a member of the
transforming growth factor-beta gene family, which mediates
male sexual differentiation. It has been shown that AMH is
positively associated with breast cancer risk.38 In this study,
AMH demonstrated a significant relationship with COAD.
Therefore, AMH may affect the progression of COAD. It was
predicted that quercetin can reduce the level of AMH and thus
lower the risk of COAD by down-regulating the expression of
the AMH gene.
According to the function enrichment analyses, the poss-
ible mechanisms for quercetin to treat PAAD, COAD, and
READ can be analyzed in the following manner. The identified
GO terms are associated with the immune response in PAAD
and the metabolism of glutathione (GSH) in COAD. GSH and
enzymes related to this antioxidant molecule are often over-
expressed in tumor cells and may contribute towards drug re-
sistance. Blockade of glutathione transferases (GSTs) has been
proposed to potentiate the efficacy of chemotherapeutic agents
in cancers. The depletion of the intracellular GSH level is
highly desirable for targeted-therapeutic agents to reverse
tumor drug resistance.39 It was predicted that quercetin can
regulate GST and affect the progression of COAD. In READ, the
identified GO terms are associated with oxidation–reduction.
It has been reported that the balance of redox reactions always
affects cell growth and apoptosis.40 It can be predicted that
quercetin may be helpful in contributing toward the balance
of redox reactions, may affect growth and apoptosis, and may
finally affect the progression of READ.
Overall, our study found new targets and mechanisms for
quercetin to inhibit digestive system cancers, which have not
been reported before. CD36 was a target of quercetin to inhibit
This journal is © The Royal Society of Chemistry 2019
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Paper
PAAD by increasing the expression of CD36 and stimulating
the immune response to prevent cancer. SLCO1B1 was identi-
fied as a target of quercetin to inhibit READ by up-regulating
SLCO1B1 to influence the uptake of drugs used for curing and
preventing READ. In COAD, down-regulation of ACADS,
ALDH3B2, UGT2A3, CDKN2A, MAF, FOSL1 and CFL2 and up-
regulation of CYP3A4 might be responsible for the anticancer
activity of quercetin towards READ.
Funding
National Natural Science Foundation of China (grant no.
31701722), Modern Agricultural Industry Technology System
(CARS-30), National Key R&D Program of China
(2017YFE0105300), Key Research and Development Plan of
Shaanxi Province (2017ZDXL-NY-0304 and 2019ZDLNY01-02-
02), China Postdoctoral Science Foundation (No.
2017M620471), Shaanxi Provincial Natural Science Foundation
(No. 2018JQ3054), and Postdoctoral Research Project of
Shaanxi Province (No. 2017BSHEDZZ103).
Conflicts of interest
There are no conflicts of interest to declare.
Acknowledgements
We would like to thank the Laboratory Center in School of Life
Sciences, Northwestern Polytechnical University for providing
the facilities to perform this study.
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