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ISSN 0012-4966, Doklady Biological Sciences, 2025. © The Author(s), 2025. This article is an open access publication.
Thioridazine Induces Increase in Expression
of the Pyruvate Transporter MPC1 Associated
with Immune Infiltration in Malignant Tumors
E. A. Bogomolova
a,
* (
ORCID: 0009-0009-9407-613
), M. M. Murashko
a,c
(
ORCID: 0000-0001-7235-5052
),
E. M. Stasevich
a,b,c
(
ORCID: 0000-0003-2150-9458
), A. N. Uvarova
a,b
(
ORCID: 0000-0002-3168-6872
),
E. A. Zheremyan
a,b
(
ORCID: 0000-0002-1230-6168
), K. V. Korneev
a,b
(
ORCID: 0000-0003-3570-8327
),
Corresponding Member of the RAS
D. V. Kuprash
a,b
(
ORCID: 0000-0002-1488-4148
),
and D. E. Demin
a,b,
** (
ORCID: 0000-0002-5553-6588
)
Received September 15, 2024; revised November 15, 2024; accepted November 20, 2024
Abstract—The MPC1 gene is involved in the transport of pyruvate into mitochondria, playing an important
role in metabolic processes. Recently, it has been reported that higher MPC1 expression correlates with an
increased number of immune cells in human cervical and lung cancers, indicating an enhanced antitumor
immune response. Reduced MPC1 levels in gastric tumors are associated with a more severe disease course.
Correlational analysis of the MPC1 gene in human lung, hippocampus and frontal cortex tissue samples based
on data from the GTEx database revealed associations of this gene with schizophrenia, non-small cell lung
cancer, and immune diseases. Our experiments showed that the mRNA level of the MPC1 gene in the non-
small cell lung cancer cell line A549 increases 5-fold under the inf luence of the schizophrenia neuroleptic thi-
oridazine. The observed elevation of MPC1 level may cause tumor infiltration by immune cells, complement-
ing the previously reported data indicating the ability of thioridazine to slow cell growth, induce apoptosis and
reduce the ability of cells to migrate.
Keywords: non-small cell lung cancer, MPC1, thioridazine, schizophrenia, immune infiltration
DOI: 10.1134/S001249662460060X
INTRODUCTION
Non-small cell lung cancer (NSCLC) accounts for
about 85% of all cases of malignant lung tumors [1].
The immunologic environment of the tumor plays a
key role in cancer development and progression as well
as response to therapy. The introduction of immune
checkpoint inhibitors, such as CTLA-4 and PD-
1/PD-L1 blockers, has been a breakthrough in the
treatment of non-small cell lung cancer. However,
many patients do not respond to treatment or show
disease progression. This highlights the need to
develop new biomarkers and therapeutic approaches
based on the interaction between the tumor and the
immune system [2].
The MPC1 gene plays a key role in the transport of
pyruvate across the inner mitochondrial membrane,
ensuring its participation in oxidative phosphorylation
and energy metabolism of the cell. Recent studies have
shown that increased expression levels of the MPC1
gene are associated with the infiltration of immune
cells into lung cancer tumor tissue. In particular, there
is a correlation between MPC1 level and the number of
follicular T-helper cells and eosinophils, which may be
related to the effectiveness of the antitumor immune
response [3]. In addition, correlation of MPC1 expres-
sion with immune infiltration is found in cervical can-
cer [4]. Reduced MPC1 levels are associated with poor
prognosis and tumor progression in gastric cancer [5].
The mitochondrial pyruvate transporter complex
(MPC) is known to consist of MPC1 and MPC2 sub-
units, and loss of either of them leads to dysfunction of
the entire complex. At the same time, the single nucle-
otide polymorphism rs10489202 in an intron of the
aLaboratory of Intracellular Signaling in Health and Disease,
Engelhardt Institute of Molecular Biology, Russian Academy
of Sciences, Moscow, 119991 Russia
bCenter for Precision Genome Editing and Genetic
Technologies for Biomedicine, Engelhardt Institute
of Molecular Biology, Russian Academy of Sciences,
Moscow, 119991 Russia
cDepartment of Molecular and Biological Physics,
Moscow Institute of Physics and Technology,
Dolgoprudny, Moscow oblast, 141701 Russia
*e-mail: elvina.elochka@gmail.com
**e-mail: denisdeminbio@gmail.com
2
DOKLADY BIOLOGICAL SCIENCES 2025
BOGOMOLOVA et al.
MPC2 gene is associated with schizophrenia in East
Asian populations [6].
In recent years, the antipsychotic drug thiori-
dazine, used in the treatment of schizophrenia, has
attracted the attention of researchers as a potential
antitumor agent, especially against various cancers,
including lung cancer [7]. The ability of thioridazine
to influence inflammation by blocking IKKβ kinase,
which in turn prevents the activation of the transcrip-
tion factor NF-κB, has been shown [8]. This drug is
able to induce apoptosis, inhibit cancer cell prolifera-
tion and migration, and selectively destroy cancer
stem cells, making it an interesting candidate to target
recurrent and resistant forms of cancer [9]. The effects
of thioridazine against cancer cells are related to the
activation of mitochondrial apoptosis through inhibi-
tion of fatty acid oxidation. It causes a decrease in
mitochondrial membrane potential, caspase-9 activa-
tion, increased Bax protein levels and decreased Bcl-2
levels, leading to cell death [7]. Thioridazine adminis-
tration is associated with a decreased risk of gastric
cancer [10].
In this study, we analyzed the mRNA expression of
MPC1 gene in human lung tissue s amples based on the
GTEx database. Based on the data available in the lit-
erature on the effect of thioridazine on mitochondrial
apoptosis, we studied the effect of thioridazine on
MPC1 gene expression in the non-small cell lung can-
cer cell line A549.
MATERIALS AND METHODS
Cells lines. The A549 non-small cell lung cancer
cell line was used in the study. Cells were cultured in
DMEM medium containing 4.5 g/L glucose (Pan-
Eco), supplemented with a mixture of antibiotics,
including penicillin (100 U/mL) and streptomycin
(100 μg/mL) (PanEco), 1% non-essential amino acids
solution (PanEco), 10 mM HEPES (GIBCO), and
10% fetal bovine serum (Biosera). Thioridazine
(PHARMACEUTICAL WORKS JELFA S.A.) was
added to the cells at concentrations of 10, 20, 30 or
40 μM, while the control samples received an equiva-
lent volume of culture medium.
Correlational analysis of the MPC1 gene. To id en-
tify the biological processes involving the MPC1 gene,
a correlation analysis was performed as described in
[11]. Briefly, RNA sequencing data from normal
human tissues were used from the GTEx database:578
lung samples, 197 hippocampus samples, and 209
frontal cortex samples (BA9). The Spearman correla-
tion of gene expression for all genes with MPC1
expression was calculated for these tissues. The top
1000 genes with the highest absolute correlation coef-
ficients were then used for gene set enrichment analy-
sis using Metascape and EnrichR.
RNA isolation and quantitative polymerase chain
reaction. Total RNA was isolated from cells 24 h after
transfection using ExtractRNA reagent (Eurogen)
according to the protocol recommended in the kit.
cDNA from total RNA was prepared with the MMLV
RT kit (Eurogen) using oligo-dT primers and random
nucleotide primers in a 1-to-1 ratio as described in
[12]. Expression analysis was performed by quantita-
tive polymerase chain reaction (qPCR) on a CFX96
Touch real-time PCR instrument (Bio-Rad Laborato-
ries) using qPCRmix-HS SYBR reagents (Eurogen) and
specific primers: for MPC1 gene—GGACTATGTC-
CGAAGCAAGG; AAATGTCATCCGCCCACTGA.
Normalization was performed for the beta-actin
ACTB gene—ACTGGGGACGACGACATGGA-
GAGAAA; GGCGTACAGGGATAGGATAGCA-
CAG. The cDNA obtained from 100 ng of isolated
RNA was used per triplicate quantitative PCR.
RESULTS AND DISCUSSION
MPC1 mRNA Expression in Human Lung Tissue
Correlates with Genes from Groups Associated with
Schizophrenia, Lung Cancer, and Immunopathologies
As a result of the gene set enrichment analysis
based on the list of one thousand genes most strongly
correlated with MPC1 in lung and brain tissues (hip-
pocampus and frontal cortex BA9), the biological pro-
cesses in which the MPC1 gene may be involved were
identified.
In normal human lung tissues, processes poten-
tially associated with MPC1 include mitochondrial
transport, amide metabolism, valine, leucine, and iso-
leucine degradation, proton transmembrane trans-
port, membrane organization, lipid biosynthetic pro-
cess, cellular catabolic process, mitochondrial protein
degradation, and aerobic respiration and respiratory
electron transport (Fig. 1a).
In addition to understanding common processes,
the association with biological processes from the
DisGeNet database that are associated with diseases is
also of interest. We identified a variety of potential
associations between the MPC1 gene and groups, and
selected groups that are associated with immune dis-
eases and lung cancer to show in the figure. Figure 1b
displays the groups associated with immune diseases:
immunodeficiencies, autoimmune diseases, neuro-
logical disorders, and allergic reactions. These results
suggest a possible link between MPC1 and immune
regulation and the pathogenesis of various immuno-
logic conditions.
Figure 1b displays the groups associated with lung
cancer: non-small cell lung cancer, its advanced and
recurrent forms, and stages III and IV. Tumor types
such as primary adenocarcinoma of the lung, squa-
mous cell non-small cell lung cancer, and various car-
cinomas of the lung were also identified. The observed
associations are in good agreement with the data on
DOKLADY BIOLOGICAL SCIENCES 2025
THIORIDAZINE INDUCES INCREASE IN EXPRESSION 3
the prognostic significance of MPC1 in lung cancer
[13] and its possible role in pathogenesis [14].
The formation of the MPC1 and MPC2 protein
complex, as well as the association of MPC2 with
schizophrenia, suggests in turn that the MPC1 gene is
related to this psychiatric disorder. Enrichment analy-
sis of functional gene groups in brain tissues associated
with schizophrenia found that for hippocampal sam-
ples, schizophrenia is in the top 3, and in frontal lobe
(BA9) samples in a list of the 15 most enriched groups.
The results of the correlation analysis emphasize
the role of MPC1 in metabolic processes and mainte-
nance of lung tissue homeostasis, and suggest an asso-
ciation with schizophrenia in brain tissue. Thus, the
results obtained in conjunction with the literature
indicate that MPC1 can potentially be considered as a
biomarker associated with the development of lung
cancer, immunopathologies and schizophrenia.
Thioridazine Increases the mRNA Expression Level
of the MPC1 Gene
It was previously shown that the antipsychotic thi-
oridazine used in schizophrenia slows growth and
reduces the ability of non-small cell lung cancer cells
Fig. 1. Genes correlating with MPC1 in lung tissues enrich functional groups associated with metabolic processes (a), immuno-
pathologies (b), and lung cancer (c). The figure shows groups of genes obtained as a result of correlation analysis of mRNA MPC1
expression in human lung tissue samples. (a) The strongest enrichment of functional groups obtained using the Metascape ser-
vice. Enrichment of groups of genes from the DisGeNET database related to immune diseases (b) and lung cancer (c).
0510
2 4
Functional groups
Functional groups
15 20 25 30
(a)
(b) (c)
35
log10(P)
log10(P)log10(P)
1.5 2.0
4
DOKLADY BIOLOGICAL SCIENCES 2025
BOGOMOLOVA et al.
to migrate, and at higher concentrations leads to apop-
tosis [15]. Our results showed that at a thioridazine
concentration of 10 μM, no significant increase in
MPC1 gene mRNA expression was observed. At a
concentration of 20 μM, a significant increase of 5 ±
2-fold was already seen, further at concentrations of
30 and 40 μM, a marked increase in MPC1 mRNA
levels of 5 ± 3 and 5.9 ± 1.4-fold, respectively, was also
found (Fig. 2).
Therapeutic dosages of thioridazine in the treat-
ment of various psychiatric disorders such as schizo-
phrenia are up to 400 mg/day, with concentrations on
the order of 10 μM observed in human serum at
400 mg/day dosage [16, 17]. Therefore, the relevance
of this cell model to the therapeutic use of thioridazine
requires a separate investigation. However, in clinical
studies, the required concentrations of the drug are
often several times lower than those used in vitro [18].
It should be noted that previous studies have found a
correlation between increased MPC1 expression and
infiltration of lung tumors by follicular T-helper cells
and eosinophils [8], and infiltration of non-small cell
lung cancer by follicular T-helper cells is associated
with a good clinical prognosis [19]. It can be assumed
that the increase in MPC1 during thioridazine expo-
sure may participate in the attraction of these types of
immune cells to the tumor.
FUNDING
This work was supported by the Ministry of Science and
High er Ed ucation of th e Russ ian Federation (g rant no. 075-
15-2019-1660).
ETHICS APPROVAL AND CONSENT
TO PARTICIPATE
This work does not contain any studies involving human
and animal subjects.
CONFLICT OF INTEREST
The authors of this work declare that they have no con-
flicts of interest.
OPEN ACCESS
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