Invest Clin 66(1): 78 - 88, 2025 https://doi.org/10.54817/IC.v66n1a07
Corresponding Author: Peili Li. Department of Obstetrics and Gynecology. Shanxi Fenyang Hospital. No. 186
Shengli Street, Fenyang, Shanxi 032299, China. Tel: +86 18735833856. E-mail: lipeili829@163.com
The role of 5-fluorouracil in Wnt/β-catenin
signalling in human papillomavirus-positive
cervical cancer cells.
Lifang Wen1, Xiaojun Liang2, Jie Ding3, Haijuan Zhang4 and Peili Li2
1Department of Obstetrics and Gynecology Teaching and Research, Fenyang College
of Shanxi Medical University, Fenyang, Shanxi, China.
2Department of Obstetrics and Gynecology, Shanxi Fenyang Hospital, Fenyang, Shanxi,
China.
3Department of Interventional, Shanxi Fenyang Hospital, Fenyang, Shanxi, China.
4Department of Pathology Student Science Teaching and Research Office, Fenyang
College of Shanxi Medical University, Fenyang, Shanxi, China.
Keywords: 5-fluorouracil; high-risk HPV-positive cervical cancer cells; T lymphocytes;
Wnt/β-catenin signalling pathway; apoptosis.
Abstract. Human papillomavirus (HPV) infection is a major risk factor for cer-
vical cancer, especially persistent infection with high-risk HPV. 5-fluorouracil (5-FU)
is a widely used antimetabolite chemotherapeutic agent that inhibits the prolifera-
tion of tumour cells by interfering with ribonucleic acid and deoxyribonucleic acid
synthesis; however, its mechanism of action has not been fully elucidated. This study
aimed to investigate the role of Wnt/β-catenin signalling in patients with high-risk
HPV with cervical cancer treated with 5-FU. Patients with high-risk HPV-positive
cervical cancer treated with surgery were taken as the research participants, and le-
sion tissues were collected during surgery. Human HPV-positive cervical cancer cells
were isolated and cultured in vitro by the enzyme combined digestion method, and
the obtained cells were divided into a control group, a paclitaxel group and a 5-FU
group. A 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay
was used to measure the proliferation of high-risk HPV-positive cervical cancer cells
under different treatment conditions. Western blotting was used to evaluate the
protein expression level of the Wnt/β-catenin signalling pathway in cells, and flow
cytometry was used to analyse the level of T lymphocytes in the patients’ blood. The
results of the MTT assay showed that the proliferation rate of cervical cancer cells in
the control group was significantly higher than that in the paclitaxel group and the
5-FU group at all detection time points (p<0.05). The expression levels of Wnt/β-
catenin protein in the 5-FU group were lower than those in the paclitaxel and the
control groups (p<0.05). The results of the T lymphocyte level comparison showed
that the ratios of CD3+ T cells, CD4+ T cells and CD4+/CD8+ cells affected by 5-FU
were higher than those before treatment (p<0.05). 5-fluorouracil can significantly
reduce the expression level of Wnt/β-catenin protein and increase the T lymphocyte
levels in cervical cancer cells.
The role of 5-fluorouracil in cervical cancer 79
Vol. 66(1): 78 - 88, 2025
Función del 5-fluorouracilo en la señalización Wnt/β-catenina
en células de cáncer de cuello uterino positivas al virus
del papiloma humano.
Invest Clin 2025; 66 (1): 78 – 88
Palabras clave: 5-fluorouracilo; células de cáncer de cuello uterino positivas al VPH de
alto riesgo; linfocitos T; vía de señalización Wnt/β-catenina; apoptosis.
Resumen. La infección por el virus del papiloma humano (HPV) es un
factor de riesgo importante para el cáncer de cuello uterino, especialmente la
infección persistente con HPV de alto riesgo. El 5-fluorouracilo (5-FU) es un
agente quimioterapéutico antimetabólico ampliamente utilizado que inhibe la
proliferación de las células tumorales al interferir con la síntesis de ARN y ADN,
pero su mecanismo de acción no ha sido completamente elucidado. Para inves-
tigar el papel de la señalización Wnt/β-catenina en los pacientes con cáncer de
cuello uterino de alto riesgo HPV tratados con 5-FU. Los pacientes con cáncer
de cuello uterino positivo para HPV de alto riesgo tratados con cirugía fueron
tomados como objetos de investigación, y se recolectaron tejidos lesionales
durante la cirugía. Se aislaron y cultivaron in vitro células de cáncer de cuello
uterino positivas para HPV humano mediante el método de digestión combina-
da enzimática, y las células obtenidas se dividieron en un grupo de control, un
grupo de paclitaxel y un grupo de 5-fluorouracilo. El MTT se utilizó para medir
la proliferación de las células de cáncer de cuello uterino positivas para HPV de
alto riesgo bajo diferentes condiciones de tratamiento. La técnica de western
blot se utilizó para evaluar el nivel de expresión proteica de la vía de señaliza-
ción Wnt/β-catenina en las células. La citometría de flujo se utilizó para anali-
zar el nivel de linfocitos T en la sangre del paciente. Los resultados del ensayo
MTT mostraron que la tasa de proliferación de las células de cáncer de cuello
uterino en el grupo de control fue significativamente mayor que en el grupo
de paclitaxel y el grupo de 5-fluorouracilo en todos los puntos de detección
(p<0,05). Los niveles de expresión de la proteína Wnt/β-catenina en el grupo
de 5-fluorouracilo fueron inferiores a los del grupo de paclitaxel y el grupo de
control (p<0,05). Los resultados de la comparación del nivel de linfocitos T
mostraron que las proporciones de células T CD3+, células T CD4+ y células
CD4+/CD8+ afectadas por el 5-fluorouracilo fueron más altas que antes del
tratamiento (p<0,05). El 5-fluorouracilo puede reducir significativamente el
nivel de expresión de la proteína Wnt/β-catenina y aumentar la actividad del
nivel de linfocitos T en las células de cáncer de cuello uterino.
Received: 16-12-2024 Accepted: 20-01-2025
INTRODUCTION
Cervical cancer is a common type of
malignant tumour in women worldwide and
has become a major disease that affects
and threatens the survival status of women
in various countries. The incidence of cer-
vical cancer in China has been rising in re-
cent years, with approximately 500,000 new
cases every year 1. There are many factors in
80 Wen et al.
Investigación Clínica 66(1): 2025
the development of cervical cancer, among
which human papillomavirus (HPV) infec-
tion is the main risk factor; in particular, per-
sistent infections of high-risk HPV present a
higher risk of cervical cancer 2. Although the
popularisation of the HPV vaccine and the
implementation of cervical cancer screening
programmes have reduced the incidence of
cervical cancer to a certain extent, deter-
mining how to effectively treat HPV-positive
patients or patients with cervical cancer that
has progressed to an advanced stage is still a
major problem for doctors 3.
As a widely used antimetabolite che-
motherapy drug, 5-fluorouracil (5-FU) has
been extensively used in the treatment of
a variety of solid tumours. It inhibits swell-
ing by interfering with ribonucleic acid
and deoxyribonucleic acid synthesis during
the proliferation of tumour cells; however,
its mechanism of action has not been fully
elucidated. Studies have shown 4 that 5-FU
must be maintained within a certain concen-
tration in cervical cancer, and the appropri-
ate extension of the administration time can
allow more cells to be exposed to the drugs,
which helps to increase the sensitivity of the
drug. In recent years, some scholars have
pointed out that 5-FU may exert its anti-tu-
mour effect by affecting signalling pathways
in the tumour microenvironment, such as
the Wnt/β-catenin signalling pathway. The
Wnt/β-catenin signalling pathway is a key
intracellular signalling network that plays
a central role in regulating cell fate, main-
taining tissue homeostasis and tumour de-
velopment 5. In many tumour types, aberrant
activation of the Wnt/β-catenin signalling
pathway is strongly associated with tumour
aggressiveness, drug resistance and poor
prognosis. However, research on the Wnt/β-
catenin signalling pathway has focused more
on colon cancer and oral cancer, and few
studies are related to high-risk HPV-positive
cervical cancer.
In addition, immune cells in the tumour
microenvironment, especially T lympho-
cytes, have an important impact on tumour
development and response to treatment. T
lymphocytes play an immune surveillance
role by recognising tumour antigens, and in
some cases, they can interact with tumour
cells and influence apoptosis 6. However, the
interaction between T lymphocytes and the
Wnt/β-catenin signalling pathway in HPV-
positive cervical cancer and its role in the
treatment of 5-FU is unclear 7,8.
Given this, the mechanism of action of
5-FU in the treatment of high-risk HPV-pos-
itive cervical cancer was analysed, and the
apoptosis effect of the Wnt/β-catenin signal-
ling pathway on tumour cells and their inter-
action with T lymphocytes in the process of
promoting apoptosis is discussed.
PATIENTS AND METHODS
Patients with high-risk HPV who under-
went surgery in our hospital between June
2022 and December 2023 were selected for
the experiment. A total of 78 patients with
positive cervical cancer were enrolled in the
study. All patients’ surgeries were performed
by the same team of doctors. The study was
approved by the ethics committee of the
hospital, and the patients and their families
voluntarily signed informed consent forms;
the whole experiment lasted for 2 weeks.
The inclusion criteria were as follows: (1)
patients who met the diagnostic criteria for
cervical cancer and had a histopathologi-
cally confirmed diagnosis of high-risk HPV-
positive cervical cancer 9; (2) patients with
no serious heart, lung, kidney or other vital
organ dysfunction and who were able to tol-
erate surgery and follow-up treatment; (3)
patients who had not received radiotherapy
or chemotherapy within 3 months before
and after enrolment; (4) patients who could
undergo surgical treatment; (5) patients
whose examination and treatment was com-
pleted under the guidance of doctors. The
exclusion criteria were as follows: (1) pa-
tients with a history of severe allergies, es-
pecially those who were allergic to 5-FU or
its adjuvant drugs; (2) pregnant or lactating
The role of 5-fluorouracil in cervical cancer 81
Vol. 66(1): 78 - 88, 2025
women; (3) patients with a history of other
malignant tumours or who had other malig-
nant tumours at the same time; (4) patients
with a history of mental illness or cognitive
dysfunction and could not cooperate with
the completion of surgery and return visits;
(5) patients with serious infection or im-
munodeficiency disease, which affected the
observation of the condition and the evalu-
ation of treatment effect; (6) patients with
severe coagulation dysfunction or bleeding
tendency; (7) patients who successfully com-
pleted the operation but did not cooperate
with the return visits.
Main instruments and reagents
The key chemicals and laboratory
equipment used in the experiments were
as follows: 5-FU (Hubei Hengjingrui Chemi-
cal) and paclitaxel as chemotherapy drugs
for specific biological effects; type I collage-
nase, which is used in the isolation process
of tissues or cells; Dulbecco’s modified Ea-
gle’s medium (DMEM) serves as a substrate
for cell culture, providing essential nutrients
10,11. Routine laboratory equipment – centri-
fuges for sample separation; fully automated
enzyme label analyser for rapid and accurate
determination of enzyme-linked immunosor-
bent assay (Nanjing Nuovezan Biotechnol-
ogy, Nanjing, China); CO2 cell culture in-
cubator (Shanghai Jinghong Experimental
Equipment Co., Ltd, Shanghai, China) to
provide a suitable growth environment for
cells; and a C-MAGhS10 magnetic stirrer for
mixing operations during experiments.
Culture of lesion cells
The following is a description of the
procedure that was adopted and the equip-
ment used. In a sterile environment, take
a small piece (volume 0.5–1 cm3) of surgi-
cally excised tumour tissue and place it in
a file containing 10% foetal bovine serum
(FBS) (Guangzhou Ruite Biotechnology)
with DMEM (Zhejiang Senrui Biotechnol-
ogy). Rinse thoroughly three times with
phosphate-buffered saline (PBS) (Zhejiang
Senrui Biotechnology) to remove impuri-
ties, then briefly soak in a solution contain-
ing penicillin and streptomycin to sterilise,
then rinse again with PBS. Use ophthalmic
surgical scissors to cut the tissue into fine
fragments (approximately 0.5 × 0.5 mm
per piece). Subsequently, add 0.25% tryp-
sin solution (Zhejiang Senrui Biotechnol-
ogy) to the tissue and digest at 37°C in a
5% CO2 incubator for 30 min, until the tis-
sue is soft. After digestion, remove the upper
layer of liquid, then add 3 ml of 0.2% col-
lagenase type I (Zhejiang Senrui Biotech-
nology) and continue digestion under the
same conditions for 30–60 min to ensure
complete tissue breakdown. Neutralisation
is performed using DMEM with 10% FBS and
filtered through a 200 mesh sieve. The col-
lected cells are further ground through a 10
ml syringe core tube to form a single-cell
suspension. After 5 min of centrifugation (at
a force of 1,105 g), remove the supernatant
and then resuspend the cells in DMEM con-
taining 15% FBS, adjusting the cell concen-
tration to 1 × 106 cells per mL. Cells are
seeded in 30 cm2 flasks and cultured at 37°C
and 5% CO2. After 24 h, observe cell attach-
ment using an inverted microscope (Ther-
mo Fisher), change the medium after 48 h
to remove unattached cells and change the
medium every 72 h. When cell confluency
coverage reaches 80%–90%, digestion is per-
formed using 0.25% trypsin, and subculture
is performed according to cell density. After
the culture is complete, cell identification is
performed to ensure that the obtained cells
are high-risk HPV-positive cervical cancer
cells 12,13. Cells are cultured on microscope
slides and morphological characteristics of
the cells are observed using light microsco-
py. Cervical carcinoma cells usually present
as epithelioid cells with irregular nuclei and
abundant cytoplasm.
Handling of cells
Cervical cancer cells cultured to the
third generation were randomly assigned to
three experimental groups: a control group,
82 Wen et al.
Investigación Clínica 66(1): 2025
a paclitaxel group and a 5-FU group. Con-
trol group: cells were not treated with any
drugs and were maintained under basal cul-
ture conditions. Paclitaxel group: cells were
co-cultured with 0.7 μmol/L of paclitaxel
during culture. 5-fluorouracil group: cells
were co-cultured with 9.6 μmol/L of 5-FU.
The number of cells in each group was con-
sistent, and three replicate wells were set up
in each well to ensure the reliability of the
experimental results.
Cell proliferation rate
The 3-[4,5-dimethylthiazol-2-yl]-2,5
diphenyl tetrazolium bromide (MTT) assay
was used to measure the proliferation of
high-risk HPV-positive cervical cancer cells
under different treatment conditions 5. The
experiment terminated the culture at differ-
ent time points and completed the determi-
nation of the cell proliferation rate. At the
end, 20 μg of MTT solution (5 mg/mL con-
centration) was added to each sample well
and incubated at 37°C for 4 h, followed by
removal of the medium. Approximately 150
μLof dimethyl sulfoxide was added and shak-
en at low speed for 10 min on a shaker to dis-
solve the previously formed formalised meth-
azole blue crystals; finally, the absorbance of
each well was measured at a wavelength of
492 nm.
Wnt/β-Catenin protein expression detection
Western blot was used to evaluate
the protein expression level of the Wnt/β-
catenin signalling pathway in cells 6. The
three groups of cells were added to a RIPA
lysis buffer, and lysis was performed on ice
to ensure complete rupture of the cell mem-
brane and release of intracellular proteins.
Cell debris was removed by centrifugation
(12,000 rpm, 10 min) and the supernatant
was collected to obtain protein samples. Ex-
tracted proteins were quantified using either
the bicinchoninic acid assay method or the
Bradford method, ensuring that the same
amount of protein was loaded in each sample
for comparison. Quantified protein samples
were mixed with loading buffer and heated
at 95°C for 5 min to denature. Samples were
loaded into polyacrylamide gels and subject-
ed to sodium dodecyl-sulfate polyacrylamide
gel electrophoresis. Depending on the mo-
lecular weight of the protein, the appropri-
ate concentration of gel (typically 10%–15%)
was selected. Following the completion of
electrophoresis, proteins from the gel were
transferred onto polyvinylidene difluoride
(PVDF) membranes. Following membrane
transfer, the PVDF membranes were blocked
with 5% non-fat dry milk and incubated for 1
h at room temperature to prevent nonspecif-
ic binding. The membranes were incubated
with specific primary antibodies and incu-
bated overnight at 4°C. The following day,
the membranes were washed to remove un-
bound primary antibodies and washed three
times for 5 min each using tris-buffered sa-
line with 0.1% Tween® 20 detergent buffer.
The membranes were incubated with horse-
radish peroxidase (HRP)-labelled secondary
antibodies for 1 h at room temperature. The
membranes were washed again to remove
unbound secondary antibodies. The mem-
branes were processed using a chemilumi-
nescent substrate, and HRP reacted with the
substrate to produce a detectable lumines-
cent signal. Signals were captured using an
imaging system and the images were record-
ed. Finally, the Wnt/β-catenin was developed
in the gel imaging system β⁃actin bands and
corresponding grey values, and the expres-
sion level of each group was calculated, with
β⁃actin as the internal control.
T lymphocyte level
A 4 mL sample of fasting venous blood
was drawn from patients with cervical can-
cer before and after surgery. The principle
of aseptic operation during collection was
strictly followed to avoid sample contamina-
tion. The collected blood sample was mixed
with an appropriate amount of anticoagu-
lant (heparin) to prevent blood coagulation
and ensure the smooth progress of the sub-
sequent separation process. The blood sam-
The role of 5-fluorouracil in cervical cancer 83
Vol. 66(1): 78 - 88, 2025
ple was centrifuged at a relative centrifugal
force of 1,180 g for 15 min. During centrifu-
gation, lymphocytes with lower density float
in the upper layer of the density gradient
medium, while other components with high-
er density, such as red blood cells and white
blood cells, sink in the lower layer. At the
end of centrifugation, the lymphocyte layer
located in the upper layer was carefully col-
lected to avoid disturbing other components
of the lower layer. Lymphocytes were gently
aspirated using a pipette and transferred to
a new centrifuge tube. Collected lympho-
cytes were washed with sterile PBS buffer to
remove residual density gradient media and
other impurities. After washing, centrifuga-
tion was performed again, the supernatant
was discarded and the precipitated lympho-
cytes were retained. The CD3+, CD4+, CD8+
cells and the CD4+/CD8+ ratio in blood were
analysed by flow cytometry (Beijing Boao
Jingdian Biotechnology Co., Ltd.) 15,16.
Experimental data processing and analysis
The SPSS software package was used
for statistical analysis. For the measurement
data that met the normal distribution condi-
tion, the independent samples t-test and one-
way analysis of variance method were used,
and the results obtained were measured as
the mean ± standard error. For categorical
data, the chi-squared test (χ²) was used for
analysis. A p-value of <0.05 was used to indi-
cate that the difference between the data is
statistically significant.
RESULTS
Comparison of cell proliferation rates at
different time points between the three
groups of cells
The MTT assay showed that the prolif-
eration rate of cervical cancer cells in the
paclitaxel group and the 5-FU group was
significantly lower than that in the control
group (p<0.05) at all detection time points,
indicating that the paclitaxel level was sig-
nificantly lower than that of the control
group (p<0.05). Both drug treatments with
fluorouracil can effectively inhibit the prolif-
eration ability of cervical cancer cells. In ad-
dition, the cell proliferation rate of the 5-FU
group was lower than that of the paclitaxel
group at all time points (p<0.05), indicat-
ing that 5-FU had a stronger inhibitory ef-
fect on cell proliferation (Fig. 1).
Comparison of Wnt/β-catenin in different
organisations
The protein expression levels of Wnt/β-
catenin in the 5-FU group and paclitaxel
group were lower than those in the control
group (p<0.01). The expression levels of
Wnt/β-catenin protein in the 5-FU group
were lower than those in the paclitaxel group
(p<0.01) (Fig. 2).
Comparison of T lymphocyte levels before
and after surgery in the observation group
The levels of CD3+ T cells, CD4+ T
cells, CD8+ T cells and CD4+/CD8+ ra-
tio were (68.94 ± 3.58), (43.28 ± 2.14),
(24.17 ± 2.11) and (2.02 ± 1.00) in the
patient after 5-FU treatment, respectively.
The level of all cells was significantly higher
than that before 5-FU treatment (p<0.05).
This reflects the activation and enhance-
ment of the intracellular immune response
after treatment.
Fig. 1. Comparison of cell proliferation rates at
different times. p<0.01 between different
groups, one-way ANOVA was used for statis-
tical analysis.
84 Wen et al.
Investigación Clínica 66(1): 2025
An increase in the CD4+/CD8+ ratio
indicates an increase in helper T cells rela-
tive to cytotoxic T cells. This involves a more
effective immune response. In contrast, the
level of CD8+ T cells (cytotoxic T cells) in pa-
tients after treatment was lower than before
treatment (p<0.05). These results indicated
that during the treatment, especially under
the action of 5-FU, some cytotoxic T cells
were activated and migrated into tumour
tissues, participating in the direct killing of
tumour cells. In addition, decreased levels
of CD8+ T cells may also be associated with
cell activation-induced apoptosis, a natural
phenomenon of T cells during sustained re-
sponses (Table 1 and Fig. 3).
DISCUSSION
Cervical cancer is the second most
common malignancy in women worldwide
after breast cancer, and the incidence is sig-
nificantly higher in developing countries,
accounting for approximately 15% of female
cancers 17,18. Although HPV vaccination and
screening technology can effectively reduce
and suppress the incidence of cervical can-
cer, HPV infection is still the main factor
that induces cervical cancer in women. Most
HPV infections are cleared by the body’s
autoimmune response; however, persistent
high-risk HPV infection may activate the
Wnt/β-catenin signalling pathway, which in
turn promotes the development of cervical
lesions 19. Aberrant activation of the Wnt/β-
catenin signalling pathway is closely related
to the occurrence of a variety of tumours.
This study examined how the Wnt/β-
catenin signalling pathway in 5-FU promotes
apoptosis of high-risk HPV-positive cervical
cancer cells, and the correlation of lympho-
cytes was discussed; furthermore, the role of
T lymphocytes in this process was analysed,
Fig. 2. Comparison of protein expression of cellsWnt/β-catenin in three groups. **: p<0.01, ***: p<0.001,
one-way ANOVA was used for statistical analysis.
Table 1
T lymphocyte levels in patients.
Cellular level/% n Before treatment After treatment t p
CD3+
78
58.24±4.58 68.94±3.58 17.060 <0.01
CD4+38.37±4.46 43.28±2.14 29.797 <0.01
CD8+21.49±2.12 24.17±2.11 8.090 <0.01
CD4+/CD8+1.58±2.11 2.02±1.00 7.619 <0.01
Note: 5-FU treatment; t test was used for statistical analysis.
The role of 5-fluorouracil in cervical cancer 85
Vol. 66(1): 78 - 88, 2025
which provided a new perspective for the
treatment of cervical cancer.
Broniarczyk et al. 20 showed that the ex-
pression level of Wnt/β-catenin protein was
positively correlated with the expression
level of high-risk HPV-positive cervical can-
cer tissues, and the high-risk HPV was posi-
tive. The content of Wnt/β-catenin protein
in cervical cancer tissues was significantly
higher than that in HPV-negative cervical
cancer tissues 20. The study of Li et al. 21
found that risperidone had a more obvious
pro-apoptotic effect on bone cells, and the
content of β-catenin protein was signifi-
cantly reduced, which may be because ris-
peridone has a certain inhibitory effect on
β-catenin protein and slows down β-catenin.
Proteins are transferred to the inner core of
the cell, which upsets the balance between
anti-apoptotic and pro-apoptotic protein
cells 21. The results showed that the expres-
sion level of Wnt/β-catenin protein in the
paclitaxel group was higher than that in the
control group after 5-FU treatment, while
the expression level of Wnt/β-catenin pro-
tein in the paclitaxel group was significant-
ly higher than that in the control group,
and there was a significant difference. This
indicates that the Wnt/β-catenin signalling
pathway may be affected by different drugs,
among which 5-FU has the most significant
effect, and 5-FU can more effectively inhibit
Wnt/β-catenin signalling pathways to en-
hance apoptosis-inducing effects on cervi-
cal cancer cells.
Shu et al. 22 found that after high-dose
5-FU treatment, the proliferation ability
and migration ability of colorectal cancer
stem cells were positively improved, while
the apoptosis rate decreased significantly.
These results suggest that 5-FU can induce
activated stem cells by activating Wnt/β-
catenin signalling cells and ultimately in-
duce the occurrence and development of
tumour diseases, which is very unfavourable
to patients 22. The study used flow cytom-
etry to analyse the level of T lymphocytes
in patients’ blood and found that under the
action of 5-FU, some cytotoxic T cells can
be activated and migrate to tumour tissues,
directly killing tumour cells, which is simi-
lar to the report of Shu et al.22 However, it
also directly shows that the decrease of T
cell level has a certain correlation with cell
Fig. 3. Lymphocyte flow cytometry Before and after surgery. A. After surgery; B. Before surgery.
86 Wen et al.
Investigación Clínica 66(1): 2025
activation inducing apoptosis, and 5-FU can
not only promote cell proliferation but also
promote apoptosis. Zheng et al. 23 proposed
that an Artemisia annua drug could affect
and inhibit the proliferation and migration
of melanoma cells by inhibiting the Wnt/β-
catenin signalling pathway and ultimately
induce apoptosis 23.
There are some limitations in this
study; limited data were used, and the cor-
relation between Wnt/β-catenin protein ex-
pression levels and T lymphocytes was not
explored. Future studies need to expand the
range of experimental data further to ex-
plore the related mechanisms of 5-FU and
Wnt/β-catenin signalling in the occurrence
and development of cervical cancer. Taken
together, the findings support the hypothe-
sis of a close link between the Wnt/β-catenin
signalling pathway and T lymphocyte levels
and reveal that 5-FU may enhance the im-
mune response in patients with cervical can-
cer by modulating this signalling pathway;
this offers new perspectives for the person-
alised treatment of high-risk HPV-positive
cervical cancer.
ACKNOWLEDGMENTS
No funding or sponsorship was received
for this study or publication of this article.
Funding
This research was supported by Shanxi
Provincial Colleges and Universities Science
and Technology Innovation Plan Project
(Project No.: 2022L673 and 2022L179); Lu-
liang City Social Development Field Key R&D
Plan Project (Project No.: 2021SHFZ49).
Ethics approval and consent to participate
This study was conducted in accordance
with the declaration of Helsinki.This study
was conducted with approval from the Eth-
ics Committee of The 305 Hospital of PLA.
Written informed consent was obtained from
all participants.
Competing interest
The authors declare that they have no
competing interests.
Availability of data and materials
All data generated or analyzed during
this study are included in this published ar-
ticle.
ORCID numbers of authors
• Lifang Wen:
0009-0000-7210-8334
• Xiaojun Liang:
0009-0005-2148-7342
• Jie Ding:
0009-0005-1274-5557
• Haijuan Zhang:
0009-0003-9095-3437
• Peili Li:
0009-0002-6957-4486
Author Contributions
Conception and design:WC, XL; Admin-
istrative support: JD, HZ; Provision of study
materials or patients: PL, LW; Collection
and assembly of data: LW, JD;Data analysis
and interpretation: PL, XL, HZ; Manuscript
writing: All authors; Final approval of manu-
script: All authors
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