TRP genes family expression in colorectal cancer
Colorectal cancer (CRC) is the most common cancer of the gastrointestinal tract. Different factors are responsible for the development of CRC. Transient Receptor Potential (TRP) which is an important component of calcium channel is associated with several pathological conditions like cancer, neurode...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
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| Цитувати: | TRP genes family expression in colorectal cancer / Y. Sozucan, M.E. Kalender, I. Sari, A. Suner, S. Oztuzcu, K. Arman, O. Yumrutas, I. Bozgeyik, B. Cengiz, Y.Z. Igci, O. Balakan, C. Camci // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 208-212. — Бібліогр.: 31 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| id |
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Sozucan, Y. Kalender, M.E. Sari, I. Suner, A. Oztuzcu, S. Arman, K. Yumrutas, O. Bozgeyik, I. Cengiz, B. Balakan, O. Camci, C. Igci, Y.Z. 2019-01-22T12:59:50Z 2019-01-22T12:59:50Z 2015 TRP genes family expression in colorectal cancer / Y. Sozucan, M.E. Kalender, I. Sari, A. Suner, S. Oztuzcu, K. Arman, O. Yumrutas, I. Bozgeyik, B. Cengiz, Y.Z. Igci, O. Balakan, C. Camci // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 208-212. — Бібліогр.: 31 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/145490 Colorectal cancer (CRC) is the most common cancer of the gastrointestinal tract. Different factors are responsible for the development of CRC. Transient Receptor Potential (TRP) which is an important component of calcium channel is associated with several pathological conditions like cancer, neurodegenerative and cardiovascular diseases. Thirty members of the family of TRP ion channel in mammals have been determined till now. The aim of this study is to investigate TRPM, TRPV and TRPC gene expression levels in tumor tissues of CRC patients and to analyze the relationship of expression in tumor tissue of CRC with other known prognostic factors. Material and Methods: In this study, 93 CRC patients were included. The level of TRP gene expression in paraffin blocks of normal and cancerous colorectal tissue samples were studied at the level of mRNA with Real-time PCR. Results: The mRNA expression level of TRPV3, TRPV4, TRPV5, TRPM4 and TRPC6 genes in 37 female and 56 male patients diagnosed with CRC was revealed lower in tumor tissue as compared to normal tissue (p < 0.05). No statistically significant differences of mRNA expression levels of other TRP genes were found. Conclusions: TRP gene family like TRPV3, TRPV4, TRPV5, TRPM4 and TRPC6 may be thought as potential genes contributing to tumorigenesis as their expression decreases in CRC as compared to normal tissues. Key Words: colorectal cancer, TRP genes family: TRPV3, TRPV4, TRPV5, TRPM4, TRPC6, mRNA expression. Kaifee Arman is a recipient of Graduate Scholarship from TUBITAK under the program 2215 — Graduate Scholarship Programme for International Students. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions TRP genes family expression in colorectal cancer Article published earlier |
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TRP genes family expression in colorectal cancer |
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TRP genes family expression in colorectal cancer Sozucan, Y. Kalender, M.E. Sari, I. Suner, A. Oztuzcu, S. Arman, K. Yumrutas, O. Bozgeyik, I. Cengiz, B. Balakan, O. Camci, C. Igci, Y.Z. Original contributions |
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TRP genes family expression in colorectal cancer |
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TRP genes family expression in colorectal cancer |
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TRP genes family expression in colorectal cancer |
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TRP genes family expression in colorectal cancer |
| title_sort |
trp genes family expression in colorectal cancer |
| author |
Sozucan, Y. Kalender, M.E. Sari, I. Suner, A. Oztuzcu, S. Arman, K. Yumrutas, O. Bozgeyik, I. Cengiz, B. Balakan, O. Camci, C. Igci, Y.Z. |
| author_facet |
Sozucan, Y. Kalender, M.E. Sari, I. Suner, A. Oztuzcu, S. Arman, K. Yumrutas, O. Bozgeyik, I. Cengiz, B. Balakan, O. Camci, C. Igci, Y.Z. |
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Original contributions |
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Original contributions |
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2015 |
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English |
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Experimental Oncology |
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Article |
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Colorectal cancer (CRC) is the most common cancer of the gastrointestinal tract. Different factors are responsible for the development of CRC. Transient Receptor Potential (TRP) which is an important component of calcium channel is associated with several pathological conditions like cancer, neurodegenerative and cardiovascular diseases. Thirty members of the family of TRP ion channel in mammals have been determined till now. The aim of this study is to investigate TRPM, TRPV and TRPC gene expression levels in tumor tissues of CRC patients and to analyze the relationship of expression in tumor tissue of CRC with other known prognostic factors. Material and Methods: In this study, 93 CRC patients were included. The level of TRP gene expression in paraffin blocks of normal and cancerous colorectal tissue samples were studied at the level of mRNA with Real-time PCR. Results: The mRNA expression level of TRPV3, TRPV4, TRPV5, TRPM4 and TRPC6 genes in 37 female and 56 male patients diagnosed with CRC was revealed lower in tumor tissue as compared to normal tissue (p < 0.05). No statistically significant differences of mRNA expression levels of other TRP genes were found. Conclusions: TRP gene family like TRPV3, TRPV4, TRPV5, TRPM4 and TRPC6 may be thought as potential genes contributing to tumorigenesis as their expression decreases in CRC as compared to normal tissues. Key Words: colorectal cancer, TRP genes family: TRPV3, TRPV4, TRPV5, TRPM4, TRPC6, mRNA expression.
|
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1812-9269 |
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https://nasplib.isofts.kiev.ua/handle/123456789/145490 |
| citation_txt |
TRP genes family expression in colorectal cancer / Y. Sozucan, M.E. Kalender, I. Sari, A. Suner, S. Oztuzcu, K. Arman, O. Yumrutas, I. Bozgeyik, B. Cengiz, Y.Z. Igci, O. Balakan, C. Camci // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 208-212. — Бібліогр.: 31 назв. — англ. |
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208 Experimental Oncology 37, 208–212, 2015 (September)
TRP GENES FAMILY EXPRESSION IN COLORECTAL CANCER
Y. Sozucan1, M.E. Kalender1,*, I. Sari2, A. Suner1, S. Oztuzcu3, K. Arman3,
O. Yumrutas 4, I. Bozgeyik4, B. Cengiz5, Y.Z. Igci3, O. Balakan6, C. Camci1
1Department of Medical Oncology, Faculty of Medicine, Gaziantep University, Gaziantep TR-27310, Turkey
2Department of Pathology, Faculty of Medicine, Gaziantep University, Gaziantep TR-27310, Turkey
3Department of Medical Biology, Faculty of Medicine, Gaziantep University, Gaziantep TR-27310, Turkey
4Department of Medical Biology, Faculty of Medicine, Adiyaman University, Adiyaman TR-02040, Turkey
5Department of Medical Biology, Faculty of Medicine, Gazi University, Ankara TR-06560, Turkey
6Department of Medical Oncology, Faculty of Medicine, Sutcu Imam University, Kahramanmaras, TR-46100, Turkey
Colorectal cancer (CRC) is the most common cancer of the gastrointestinal tract. Different factors are responsible for the development
of CRC. Transient Receptor Potential (TRP) which is an important component of calcium channel is associated with several patho-
logical conditions like cancer, neurodegenerative and cardiovascular diseases. Thirty members of the family of TRP ion channel
in mammals have been determined till now. The aim of this study is to investigate TRPM, TRPV and TRPC gene expression levels
in tumor tissues of CRC patients and to analyze the relationship of expression in tumor tissue of CRC with other known prognostic
factors. Material and Methods: In this study, 93 CRC patients were included. The level of TRP gene expression in paraffin blocks
of normal and cancerous colorectal tissue samples were studied at the level of mRNA with Real-time PCR. Results: The mRNA expres-
sion level of TRPV3, TRPV4, TRPV5, TRPM4 and TRPC6 genes in 37 female and 56 male patients diagnosed with CRC was revealed
lower in tumor tissue as compared to normal tissue (p < 0.05). No statistically significant differences of mRNA expression levels
of other TRP genes were found. Conclusions: TRP gene family like TRPV3, TRPV4, TRPV5, TRPM4 and TRPC6 may be thought
as potential genes contributing to tumorigenesis as their expression decreases in CRC as compared to normal tissues.
Key Words: colorectal cancer, TRP genes family: TRPV3, TRPV4, TRPV5, TRPM4, TRPC6, mRNA expression.
Colorectal cancer (CRC) is the most common can-
cer of the gastrointestinal tract. When evaluated along
with rectal cancer, it ranks third as the most common
cancer being observed after prostate and lung cancer
in males and breast and lung cancer in females.
Genetic and environmental factors have an in-
fluential role in the development of CRC. Colorectal
cancers are sporadic cancers even though the genetic
predisposition is the most prominent risk factor in the
majority of CRC. Cellular oncogenes, growth factors
and receptors play a pivotal role in the development
and growth of the CRC [1]. Alike other types of cancer,
the development and progression of CRC also results
from multiple genetic variations.
In most of the recently conducted studies, it has
been found that that the ion channels play a critical
role in cell proliferation. Blockade of these channels
have been found to inhibit the development of cancer
cells. Therefore ion channels have received a consi-
derable attention for therapeutic targets or prognostic
biomarkers [2].
Being an ion channel, the family of Transient
Receptor Potential (TRP) channels also comprise
of seven different subsets. These are TRPC (ca-
nonical), TRPV (vanilloid), TRPM (melastatin), TRPML
(mucolip), TRPP (polycys pyridine), TRPE (ankyrin
transmembrane protein) and TRPN (nompc-like) which
are associated with many pathological and physiologi-
cal functions. The relationship between TRP channels
and cancer suggests that TRP is effective in many
ongoing cancer processes like tumor invasion, mi-
gration, angioge nesis, apoptosis, differentiation and
proliferation [3].
TRPM8, TRPM1, and TRPV6 are highly expressed
in cancer cells and the amount of protein being
expressed changes with progression from normal
to tumorigenic and then to metastatic cells. The ex-
pression levels of some other TRP channels, including
TRPC1, TRPC6, TRPM5 and TRPV1 is also increased
in cancer tissues [4]. In a recent study, TRPC5 was
found to be overproduced at the mRNA and protein
levels in 5-Fu-resistant human CRC cells [5]. TRPM8,
TRPM1 and TRPV1 are potential diagnostic markers
for the prognosis of tumor development especially the
degree of tumor aggression, and are potential targets
for pharmaceutical interventions [4].
In this study, our aim is to find out gene expression
levels of TRPM, TRPV, TRPC in cancerous tissues
of colorectal cancer patients. We also target to analyze
the relationship of expression in tumor tissue of CRC
with other known prognostic factors like age, gender,
stages of cancer, location and size of tumor, vascular
invasion, perineural invasion, lymph node involvement
and to investigate the usability as a therapeutic target.
MATERIALS AND METHODS
Selection of Patients. This study was performed
during 2001–2010 and was undertaken in correlation
with Department of Medical Oncology, Medical Faculty
Hospital, Gaziantep University which was responsible
for patients follow-up and their treatment and De-
partment of Pathology, Medical Faculty, Gaziantep
Submitted: April 22, 2015.
*Correspondence: E-mail: kalender@gantep.edu.tr
Abbreviation used: CRC — colorectal cancer; TRP — transient re-
ceptor potential.
Exp Oncol 2015
37, 3, 208–212
Experimental Oncology 37, 208–212, 2015 (September) 209
University which carried out pathological testing for
different stages of colon cancer. This study was car-
ried out on 96 patients for which approval of the local
ethics committee have been made retrospectively.
Three out of 96 paraffin blocks of cancer tissue could
not later be found and thus were excluded from the
study. As a result, 93 cases of colon cancer were in-
cluded in this study.
The study of the patient demographics (age, gen-
der) and their different pathological features (tumor
location, tumor size, histological type, differentiation,
vascular and perineural invasion, involvement of lymph
node), their different clinical stages, follow-up and their
survival span have been registered in a standard form.
The stored paraffin blocks of 93 patients were ob-
tained from the archives of Department of Pathology,
Faculty of Medicine, Gaziantep University for further
study. The obtained samples were revaluated under
light microscope. For each subject, separate paraffin
blocks containing normal colon mucosa and infected
colon cancerous tissue slides were selected.
RNA isolation from FFPE tissue. The total RNA
was obtained from samples embedded in paraffin
using RNeasy FFPE Kit (Qiagen Sample and Assay
Technologies, Hilden, Germany, Cat. No: 73504).
The quantity and integrity of the RNA was measured
using Epoch Microplate Spectrophotometer (BioTek
Instruments, Winooski, USA). After total RNA isolation
and purification, samples were diluted to a concentra-
tion 50 ng/μl and stored at ‒80 °C until further use.
RNA was converted to cDNA using Ipsogen Reverse
Transcription-Dx Kit (Qiagen, Hilden, Germany, Lot
no: 11–11–11) following the manufacturer’s protocol.
qPCR. Gene expression analysis was done on a Bio-
Mark™ HD System (Fluidigm Corporation, CA, USA).
Samples were preamplified for 14 cycles using TaqMan®
PreAmp Master Mix (Life Technologies, Foster City, CA,
USA, Cat. No: 4391128) prior to real-time qPCR. Real-
time qPCR was done in BioMark 96.96 Dynamic Array
(Fluidigm Corporation, CA, USA) using a set of TaqMan
Gene Expression Assays (Life Technologies, Foster City,
CA, USA, Cat. No: 4333458).
Statistical analysis. Real-time PCR was per-
formed using Fluidigm Dynamic Array along with
BioMARK HD System and the gene expression mea-
surements and their data analysis was evaluated using
Biogazel and qBasePLUS software program. ACTB
was used as housekeeping gene for the normalization
of the target genes.
Significance level was considered as p < 0.05 using
Mann — Whitney test to analyze data. The dispersion
values were also taken into account during compari-
son of Mann — Whitney average test values. When
“datapoints” corresponding to the sample number
measured in the assay fall below 12, significance test
became quite sensitive and apart from the means,
the distribution analysed for statistical significance
of the test was probably found insufficient to gene-
rate a standard curve. Therefore, p value expressing
the significance level of the test would be very high
in this group and a significant comparison would not
be expressed statistically.
RESULTS
A total of 93 patients including 37 women and
56 men who were pathologically diagnosed with CRC
were included in study. The ratio of men to women
was 1.51. The patients were in the age group of 12–
87 years and the mean age of patients was found
to be 60 ± 16.1 years. The general demographic, clinical
and pathological characteristics are shown in Table 1.
The smallest and largest tumor size was 1.5 cm and
16 cm, respectively. The mean tumor size was 5.8 ±
2.6 cm. According to the TNM classification, 9 (9.7%)
patients were in stage I, 33 (35.5%) patients were
in stage II, 26 (27.9%) patients were in stage III and the
remaining 25 (26.9%) patients were in stage IV (Fig. 1).
Table 1. Distribution of general demographic, clinical and pathological
characteristics of patients
Characteristics Number of patients, n (%)
Gender
Female 37 (39.8)
Male 56 (60.2)
Localization
Rectosigmoid 9 (9.7)
Rectum 12 (12.9)
Sigmoid 18 (19.4)
Right Colon 31 (33.3)
Left Colon 11 (11.8)
Transverse Colon 12 (12.9)
Histology
Adenocarcinoma 87 (93.5)
Mucinous carcinoma 6 (6.5)
Vascular invasion
Negative 78 (83.9)
Positive 15 (16.1)
Perineural invasion
Negative 86 (92.5)
Positive 7 (7.5)
Differentiation
Unknown 64 (68.8)
High 16 (17.2)
Medium 10 (10.8)
Less 3 (3.2)
Lymph node involvement
Negative 52 (55.9)
1–3 positive 29 (31.2)
4 and above positive 12 (12.9)
0
5
10
15
20
25
30
35
40
Stage I Stage II Stage III Stage IV
Fig. 1. Distribution of patients according to TNM staging
Real-time PCR was performed using “Fluidigm Dy-
namic Array” along with BioMARK HD System and the
gene expression measurements and their data analysis
were evaluated using Biogazel and qBasePLUS soft-
ware program. ACTB was used as housekeeping gene
for the normalization of the targets mentioned in Table 2.
210 Experimental Oncology 37, 208–212, 2015 (September)
The obtained gene expression data from normal
and cancerous tissues of the same patient were
compared (Fig. 2, Table 3). For this purpose, Real
Time PCR was carried out of the mRNA obtained from
93 patients for both tumor tissue and the normal ones.
The expression of TRPV4, TRPM4, TRPV3, TRPC6 and
TRPV5 in tumor tissues were found to have lesser gene
expression as compared to normal tissues (p < 0.05).
When expression levels of other TRP genes in tissues
were compared, any significant difference was not
found (p > 0.05).
Table 2. List of target genes to be investigated for gene expression in TRP
channels
TRPA1 TRPC6 TRPM4 TRPV1 TRPV6
TRPC1 TRPC7 TRPM5 TRPV2 ACTB
TRPC3 TRPM1 TRPM6 TRPV3 −
TRPC4 TRPM2 TRPM7 TRPV4 −
TRPC5 TRPM3 TRPM8 TRPV5 −
0.0625
0.03125
1
0.5
0.25
0.125
TRPV4 TRPM4 TRPV3 TRPC6 TRPV5
Ex
pr
es
si
on
ra
te
o
f t
um
or
/n
or
m
al
ti
ss
ue
s
Fig. 2. Expression rate of tumor/normal tissues
Table 3. Comparison of gene expression data from normal and cancero us
tissues
Target
gene
Tissue
type
Gene expression
level
Nu
m
be
r o
f p
at
ien
ts Tumor/normal ratio
of gene expression
p
No
rm
al
ize
d
m
ea
n
va
lu
e
95
%
C
I l
o-
we
r l
im
it
95
%
C
I h
i-
gh
er
li
m
it
M
ea
n
va
lu
e
95
%
C
I l
o-
we
r l
im
it
95
%
CI
h
i-
gh
er
li
m
it
TRPV4 Normal 1.000 0.656 1.524 76 0.116 0.06 0.222 3.26E-06Tumor 0.116 0.070 0.193 57
TRPM4 Normal 1.000 0.666 1.502 76 0.148 0.074 0.294 6.41E-05Tumor 0.148 0.082 0.266 47
TRPV3 Normal 1.000 0.655 1.527 73 0.169 0.085 0.336 1.28E-04Tumor 0.169 0.096 0.298 48
TRPC6 Normal 1.000 0.610 1.640 54 0.234 0.111 0.493 3.75E-03Tumor 0.234 0.132 0.415 41
TRPV5 Normal 1.000 0.651 1.536 55 0.303 0.152 0.605 1.44E-02Tumor 0.303 0.171 0.535 45
For the comparison of data related to different can-
cer stages, the number of patients in different groups
was counted. In stage I, 9 patients; in stage II, 33 pa-
tients; in stage III, 26 patients; in stage IV, 25 patients
were found out. However, for the comparison of aver-
age Mann — Whitney test values, the dispersion va-
lues should be taken into account. When “datapoints”
corresponding to the sample number measured in the
assay fall below 12, significance test became quite
sensitive and apart from the means, the distribution
analysed for statistical significance of the test was
probably found insufficient to generate a standard
curve. As the total number of patients in stage I was 9,
therefore total number of patients in stage I and II were
combined together in one group for their evaluation.
As a result, stage I + II (42 patients), stage III (26 pa-
tients), stage IV (25 patients) were evaluated.
When the gene expression of cancerous tissues for
different groups of patients for various stages as stage
I + II, stage III and stage IV were compared with each
other, there was no significant difference being found
(p > 0.05).
The gene expression data obtained from the tumor
tissues showed no significant differences statistically
for tumor localisation, histological types, perineural
invasion, vascular invasion, lymph node involvement,
differentiation grade, patients gender, tumor size,
status of recurrence and metastasis (p > 0.05).
DISCUSSION
The tumor formation involves the conversion of nor-
mal cells into hyperplastic, dysplastic, neoplastic and
finally metastatic cancerous cells. This transformation
is triggered by accumulation of certain mutated key
signalling proteins which are encoded by oncogenes
and tumor suppressor genes and as a result more
aggressive cells are formed in order to compete with
adverse local surroundings.
Expression of many proteins in cancer cells in-
creases or decreases as compared to normal cells.
Some of the proteins which are encoded by oncogenes
in majority and tumor suppressor genes play a crucial
role in tumor growth and metastasis formation. Although
the other proteins which are responsible for intracellular
Ca2+ homeostasis take role in cancer progression, they
are not associated with the development of tumor and/
or malignant cells [6–8](Rosado JA, 2004, #66).
Ca2+ is known to be a versatile carrier of cell regula-
ting information right from the formation of cell till its
death. In recent studies, the role of calcium messen-
ger has been shown in several cell processes like cell
division, cell motility, hormone secretion, metabolism,
nervous system functioning, protein turnover, gene ex-
pression, developmental regulation and programmed
cell death (apoptosis) [9–11].
Many members of the TRP family of Ca2+ and Na+
permeable channels show altered expression in can-
cer cells. The most studied proteins of TRP family are
TRPM8, TRPV6, TRPM1 and TRPV1 [12–14]. To date,
most changes involving TRP proteins do not involve
mutations in the TRP gene but rather increased or de-
creased levels of expression of the normal (wild type)
TRP protein, depending on the stage of the cancer
In a number of studies during the past 20 years,
the expression and activity of TRP channels have been
shown to change during cancer. In particular, TRPC,
TRPM and TRPV have been reported to be associated
with tumorigenesis and the growth and development
of cancer cells [4]. As a result of these findings, TRP
channel expressions have been proposed as means
of prognosis or diagnosis of some cancers and targe-
ting of TRP channels have been proposed as a novel
therapeutic strategy. For example, TRPV6 and
TRPM8 have been proposed as an indicator (marker)
Experimental Oncology 37, 208–212, 2015 (September) 211
for prostate cancer development and TRPC6 as a the-
rapeutic target for oesophageal cancer [15–17].
As there is an association between TRPM, TRPV,
TRPC of TRP family especially with malignant cell division
and its progression, therefore we targeted to find out the
gene expression levels of TRPM, TRPC and TRPV in CRC.
As far as we know, there is no literature that shows
the gene expression of the extensive TRP gene family
in CRC. Our study is the most comprehensive study
on this issue and can pave a way to future works related
to this. In literature we find only the expression study
of TRPM8, TRPV1 and TRPV6 in CRC [18–21].
In this particular study, comparative study has been
done between the gene expression data of cance-
rous tissues and normal tissues of the same patient
suffering from CRC. According to the data obtained
by using Real-time PCR reaction, the gene expression
of TRPV4, TRPM4, TRPV3, TRPC6 and TRPV5 in can-
cerous tissues were found to be lower as compared
to normal tissues (p < 0.05). When expression levels
of other TRP genes in tissues were compared, no sig-
nificant difference was found statistically (p > 0.05).
Duncan et al. conducted a research on human me-
lanoma specimens using in situ hybridisation method
and found that there was an increased TRPM1 expres-
sion in benign cells (melanocytic nevi) but there was
no decrease in expression in primary melanoma as well
as metastatic melanomas [22]. In our study, we also
noticed a decrease in synthesis of TRPV4, TRPM4,
TRPV3, TRPC6, TRPV5 in cancerous cells as compared
to normal cells. TRPM1 showed a correlation with
mRNA expression of melanocytic tumor progression,
tumor thickness and its aggressiveness. In aggressive
tumors, the mRNA expression of TRPM1 was contem-
plated to be lower or in an undetectable amount. These
observations showed TRPM1 to be a tumor suppressor
gene [23–25].
In the literature, there are no studies showing the
expression of TRPM1 in CRC. We did not find any
statistical difference of TRPM1 expression between
tumor and normal tissues. This can be explained due
to difference in tissues and diversity in cells.
Tsavaler et al. [21] investigated the relationship
of TRPM8 expression between normal prostate tissue
and prostate cancer tissue using in situ hybridisation
analysis and concluded that there was a significant in-
crease of TRPM8 expression in prostate cancer tissues
as compared to normal ones. TRPM8 expression was
also studied in a number of non prostatic primary tumors
of breast, colon, lung, and skin origin and a significant
increase in its expression was found as compared to nor-
mal tissues. But in our study, there was no statistically
significant difference of TRPM8 expression between
tumor and normal tissues. We used RT-PCR from paraf-
fin blocks as a technique while Tsavaler et al. employed
in situ hybridisation technique as method of analysis [21].
Suguro et al. [26] stated that diffuse large B-cell lym-
phoma accounts for 30% of non-Hodgkin’s lymphomas
and noticed an increase of TRPM4 mRNA expression
in CD5+. TRPM4 being one of the genes involved in the
development of a “CD5 signature”, therefore it can
be used as a prognostic clinical marker. In our study,
TRPM4 expression was significantly lower in tumor
tissue than in normal tissue. The reason for this may
be due to difference in types of tissues.
Prawitt et al. [27] showed a sharp increase
in TRPM5 mRNA expression in Wilms’ tumor and rhab-
domyosarcoma. There was no statistically significant
difference of TRPM5 expression between tumor and
normal tissues in our study.
As far as we know there has been no work done
related to expression study of TRPV4, TRPV3,
TRPV5 in CRC. We found a significant decrease
in expression of TRPV4, TRPV3, TRPV5 in cancerous
tissues than normal tissues. When compared with
normal tissues or cells, the TRPV6 mRNA expression
and/or TRPV6 protein expression shows a significant
increase in prostate cancer tissue, human colon can-
cer, breast cancer, thyroid cancer and ovarian cancer
tissues [18, 28–30]. As compared to normal tissues,
increase in TRPV6 mRNA expression is observed
in prostate cancer LNCaP and PC-3 cell line, CRC
SW480 cell line and chronic myelogenous leukaemia
K-562 cell lines [18, 19].
Immunohistochemical studies demonstrated that
TRPV1 is expressed in adenocarcinoma [20]. Extracel-
lular polyamines which are agonists of TRPV1 are pre sent
in considerable concentration in the gastrointestinal
tract and at synapses and these levels increase during
inflammation and cancer [31]. As compared to this,
our study showed no significant difference statistically
of TRPV1 expression between tumor and normal tissues.
The present study detected an increase in gene
expression of TRPM8, TRV6, TRPV1 in CRC tissues
as compared to normal tissues which makes it different
from others studies. Channel expression in tumor is per-
formed by Western blot assay, immunohistochemical
studies, RT-PCR or in situ hybridisation techniques.
In order to prevent differences in results arising due
to different operating methods, it is proposed to carry
out comparison studies of these four techniques simul-
taneously from the same tissues. Gene expression stu-
dies should be carried out from paraffin blocks or fresh
obtained tissues. We performed this study on paraffin
blocks but further studies can be carried out using fresh
tissues because ambient conditions during preparation
of paraffin blocks can be an influential factor in channel
expression studies. In addition, studies on fresh tis-
sues will certainly aid in better understanding of TRP
expression pattern in CRC because of good correlation
between mRNA and protein levels in fresh tissues.
In the present study there is markedly lower expres-
sion of TRPV4, TRPM4, TRPV3, TRPC6, TRPV5 in tu-
mor tissues of CRC than normal tissues. There may
arise some questions related to present study like
what about effectiveness of these genes in CRC, what
are the related mechanisms related to its suppres-
sion. Epigenetic factors may be involved in the lower
expression of TRPV4, TRPM4, TRPV3, TRPC6 and
TRPV5 in these results.
212 Experimental Oncology 37, 208–212, 2015 (September)
In conclusion, genes like TRPV4, TRPM4, TRPV3,
TRPC6 and TRPV5 which are showing lower expression
in CRC tissues are considered to be prominent gene
candidates for potential tumor growth. This hypothesis
needs to be supplemented and supported with further
studies for its verification. New targeted therapeutic
agents can be developed by employing TRP channel
inhibitors in further studies.
ACKNOWLEDGEMENT
Kaifee Arman is a recipient of Graduate Scholarship
from TUBITAK under the program 2215 — Graduate
Scholarship Programme for International Students.
CONFLICT OF INTEREST
Authors have no conflicts of interest regarding
the subject of this manuscript.
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