Platinum-based chemotherapy enhances SEMA5B expression in HepG2 and SW480 cancer cells

Authors

Abstract

AimChemoresistance remains a major obstacle limiting the effectiveness of cancer therapy and contributes to poor clinical outcomes. Recent evidence indicates that members of the semaphorin family are involved in tumor progression and treatment response. Sema5B has been implicated in tumor development; however, its role in the response to chemotherapy has not yet been clarified. The present study aimed to investigate the effects of cisplatin and oxaliplatin on Sema5B expression in the HepG2 hepatocellular carcinoma and SW480 colorectal cancer cell lines.
MethodsCells were treated with increasing concentrations (1–200 μM) of cisplatin or oxaliplatin for 24, 48, and 72 hours, and cell viability was assessed using the MTT assay. IC50 values were calculated, and Sema5B protein expression was evaluated by Western blot analysis following 24 and 72 hour treatments.
ResultsBoth cisplatin and oxaliplatin induced a reduction in cell viability in a time and dose-dependent manner. Moreover, treatment with these platinum-based agents resulted in a marked increase in Sema5B expression in HepG2 and SW480 cells.
ConclusionThese findings suggest that platinum-based chemotherapy can modulate Sema5B expression in a cell type dependent manner and that Sema5B may participate in adaptive cellular responses to chemotherapeutic stress. This study provides preliminary evidence supporting further investigation of Sema5B as a potential molecule associated with treatment response in cancer.

Keywords

Introduction

Cancer remains a major global health problem with a steadily increasing incidence. Despite advances in targeted therapies and personalized medicine, conventional chemotherapy remains a cornerstone of treatment for many solid tumors. Among these agents, platinum-based compounds hold a central role in clinical practice due to their broad antitumor spectrum and well-characterized mechanisms of action.¹
Platinum compounds are alkylating agents that exert cytotoxic effects mainly by forming covalent bonds with DNA.² The intra- and interstrand DNA crosslinks they generate disrupt replication and transcription, leading to cell cycle arrest and apoptosis.³ As a first-generation platinum drug, cisplatin has shown significant efficacy in several malignancies, including breast, liver, colorectal, testicular, ovarian, and lung cancers.² Oxaliplatin, a third generation platinum analogue, received FDA approval in 2002 for the treatment of colorectal cancer and is widely used, either alone or in combination regimens, in stage II/III colon cancer, metastatic colorectal cancer, and non small cell lung cancer.⁴
Although platinum-based drugs are effective against many cancers, resistance frequently develops and limits treatment success. Major mechanisms include enhanced DNA repair, altered drug transport, suppression of apoptosis, and tumor microenvironment–related adaptive responses.⁵ Therefore, identifying novel molecular regulators involved in chemoresistance is critical for improving therapeutic efficacy.
The semaphorin family consists of extracellular signaling proteins, including secreted, transmembrane, and cell surface forms.⁶ Based on their structural characteristics, they are classified into eight classes, comprising a total of 30 proteins.^6,7 The Sema protein family contains a conserved Sema domain consisting of approximately 500 amino acids, which enables both semaphorin dimerization and receptor binding.⁷ This domain is also present in plexin family proteins and several receptor tyrosine kinases. Recent studies have shown that semaphorins play important roles in cancer related processes such as cell migration, metastasis, angiogenesis, apoptosis, and immune regulation.⁶ Semaphorins may act as either tumor suppressors or tumor promoters, depending on the cancer type, tissue context, and microenvironment. Consequently, interest in their use as cancer biomarkers and therapeutic targets has been steadily increasing. In addition, some members of the semaphorin family have been reported to modulate chemotherapy sensitivity or resistance mechanisms.
Semaphorin 5B (Sema5B) is a member of class 5 semaphorins and exists in both transmembrane and GPI-anchored forms.⁸ Although Sema5B is known to play a role in the development and progression of some cancers, its role in chemotherapy response remains unclear. Platinum-based agents are known to modulate various cellular pathways and contribute to the development of drug resistance. The present study investigates the potential role of Sema5B in cellular responses to chemotherapeutic stress by examining its expression following cisplatin and oxaliplatin treatment in HepG2 hepatocellular carcinoma and SW480 colorectal cancer models.

Materials and Methods

TCGA Based Analysis of SEMA5B ExpressionGene expression analysis was performed using RNA sequencing (RNA-seq) data from The Cancer Genome Atlas (TCGA) database. SEMA5B transcript levels were compared between primary tumor and adjacent normal tissues in colon adenocarcinoma (COAD) and liver hepatocellular carcinoma (LIHC) cohorts. The COAD dataset included 275 tumor and 41 normal samples, while the LIHC dataset comprised 369 tumor and 50 normal samples. Boxplot data were obtained from GEPIA2, which integrates TCGA and GTEx datasets, and were used for differential expression analysis.
Cell cultureThe HepG2 and SW480 were obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum and 100 U/mL penicillin–streptomycin at 37 °C in a humidified atmosphere with 5% CO₂.
Ethical ApprovalThis study was conducted exclusively using in vitro cell culture models and did not involve human participants or animals; therefore, ethical approval was not required.
Cell viability assaysCells were seeded at 5 × 10³ cells per well in 96-well plates. After overnight incubation, HepG2 cells were treated with cisplatin and SW480 cells with oxaliplatin (Sigma-Aldrich, St. Louis, MO, USA) at concentrations of 1–200 µM for 24, 48, or 72 h. Cell viability was evaluated using the MTT assay. Cells were incubated with 1 mg/mL MTT for 1 h at 37 °C, after which the medium was removed and formazan crystals were dissolved in 200 µL DMSO. Absorbance was measured at 570 nm to determine relative cell viability.
Western blot analysisHepG2 and SW480 cells were seeded in T25 flasks and allowed to adhere for 24 h, followed by treatment with the IC₅₀ concentrations of cisplatin or oxaliplatin for 24 or 72 h. Cells were harvested, washed with ice-cold PBS, and lysed at 4 °C. Total protein concentrations were determined using a BCA assay. Equal amounts of protein were separated by 10% SDS–PAGE and transferred onto membranes. After blocking in TBS-T containing 0.1% Triton X-100 and non-fat dry milk, membranes were incubated with primary antibodies against Sema5B (Invitrogen, San Diego, CA, USA), followed by HRP-conjugated secondary antibodies. β-actin served as the loading control. Protein bands were detected using enhanced chemiluminescence and visualized with a chemiluminescence imaging system.
Statictical analysisStatistical analysis was performed using one-way ANOVA in GraphPad Prism 9. Data are presented as mean ± standard deviation (SD) from three independent experiments. The half-maximal inhibitory concentration (IC50) values were calculated by non-linear regression analysis. Protein expression levels were quantified by densitometric analysis using ImageJ software. The intensity of each target protein band was normalized to the corresponding β-actin band to obtain relative expression values (protein/β-actin ratio). A p value ≤ 0.05 was considered statistically significant. All experiments were performed in triplicate.
Reporting GuidelinesThis experimental in vitro study was reported in accordance with the Materials Design Analysis Reporting (MDAR) framework to ensure transparent reporting of experimental design, materials, methods, and data analysis.

Results

Platinum Agents Reduce Viability Dose-Dependently in Gastrointestinal Cancer CellsThe cytotoxic effects of cisplatin and oxaliplatin on HepG2 and SW480 cancer cell lines were evaluated by MTT assay following treatment with increasing concentrations (1–200 µM) for 24, 48, and 72 hours. Both platinum-based agents exhibited a clear dose and time dependent reduction in cell viability in the respective cell models. Cisplatin treatment markedly decreased HepG2 cell viability by 71%, 82%, and 90% at 24, 48, and 72 hours, respectively, indicating a progressive enhancement of cytotoxic activity over time (Fig. 1A). Similarly, in SW480 cells, oxaliplatin caused a significant reduction in viability, with decreases of 49%, 52%, and 68% at 24, 48, and 72 hours, respectively (Fig. 1B).
To further quantify drug sensitivity, half-maximal inhibitory concentration (IC50) values were calculated using nonlinear regression analysis. In HepG2 cells, the IC50 values for cisplatin declined substantially with prolonged exposure, measured as 27.64 µM at 24 hours, 9.2 µM at 48 hours, and 0.6 µM at 72 hours. In SW480 cells, oxaliplatin IC50 values were 164 µM, 28.7 µM, and 20.37 µM at 24, 48, and 72 hours, respectively. For further mechanistic investigations, the IC50 values determined at 24 and 72 hours were used as the experimental treatment concentrations.
SEMA5B Expression Is Upregulated in Colon and Liver Cancer TissuesAnalysis of Sema5B expression in TCGA datasets revealed a significant upregulation in tumor tissues compared with adjacent normal tissues in both colon adenocarcinoma (COAD) and liver hepatocellular carcinoma (LIHC). In COAD, tumor samples (n=275) showed moderately elevated expression relative to normal controls (n=41), whereas in LIHC, tumor samples (n=369) exhibited a more pronounced increase compared with normal tissues (n=50), with the difference reaching statistical significance. These results suggest a tumor specific upregulation of Sema5B, which appears more marked in LIHC than in COAD (Fig. 2A).
Cisplatin and Oxaliplatin Induce Sema5B Expression in HepG2 and SW480 Cancer CellsTo investigate the effects of cisplatin and oxaliplatin on Sema5B protein expression, we performed Western blot analysis. HepG2 cells were exposed to cisplatin at 27.64 µM for 24 h and 0.6 µM for 72 h, whereas SW480 cells were treated with oxaliplatin at 164 µM and 20.37 µM for 24 h and 72 h, respectively. Cisplatin markedly increased Sema5B protein expression in a time-dependent manner compared with control. In addition, oxaliplatin induced a more moderate elevation in Sema5B protein levels. these findings suggest that Sema5B may play a role in the cellular response to both cisplatin- and oxaliplatin-based therapies (Fig. 2B-C).

Discussion

Drug resistance remains one of the most critical challenges limiting the success of cancer therapy. This phenomenon not only adversely affects patient survival rates but also leads to prolonged treatment duration, unnecessary drug exposure, and inefficient utilization of medical resources. Current advances increasingly focus on elucidating the molecular mechanisms underlying drug resistance; in particular, identifying signaling pathways that are activated or suppressed during this process, targeting resistance-associated molecules for therapeutic inhibition, and defining diagnostic and complementary biomarkers that may enable early intervention.^9,10 In this context, investigating alternative regulatory protein families that shape tumor biology has gained growing importance.
Recent studies have revealed that the semaphorin protein family, initially characterized for its role in axonal guidance, also plays critical roles in cancer development and progression.^11,12 Semaphorins have been shown to directly influence tumor behavior by regulating key cellular processes such as proliferation, migration, invasion, and interactions with the tumor microenvironment.^13,14 In particular, Sema4D has been reported to enhance tumor cell proliferation, promote migration and invasion, and support angiogenesis in malignancies such as ovarian cancer and non-small cell lung cancer.^15,16 Another study demonstrated that antibody mediated blockade of Sema3A in glioblastoma inhibited tumor growth through downregulation of the PI3K/Akt signaling pathway.¹⁷ Molecular profiling analyses further indicate that Sema3C, Sema3E, Sema5A and Sema6D are upregulated in various cancer types, whereas Sema3A, Sema3B, Sema3F, and Sema3G are frequently suppressed.⁶ These distinct expression patterns suggest that the semaphorin family exerts bidirectional biological effects, functioning either as tumor suppressors or as oncogenic regulators depending on the cellular context.
Recent studies suggest that Sema5B may contribute to tumor progression in several cancer types. In renal cell carcinoma, Sema5B has been reported to play a functional role, and its inhibition suppresses tumor growth.⁸ Similarly, silencing of Sema5B resulted in reduced proliferation, invasion, and glycolytic activity in esophageal squamous cell carcinoma.¹⁸ These findings indicate that Sema5B may be associated with metabolic reprogramming and an aggressive tumor phenotype. Moreover, the overexpression of Sema5B in certain malignancies has led to its consideration as a potential oncogenic semaphorin, and it has also been proposed as a prognostic biomarker for Burkitt lymphoma.¹⁹ The TCGA-based analyses conducted in this study are consistent with the existing literature and demonstrate that Sema5B expression is significantly elevated in colorectal and liver cancer tissues relative to corresponding normal tissues. This increase suggests that Sema5B may be linked to tumor development, cellular adaptation, tissue specificity, and interactions with the tumor microenvironment.
Recent studies indicate that the semaphorin family functions not only as structural guidance molecules involved in tumor progression but also as important modulators of cellular responses to chemotherapy and the signaling networks associated with drug resistance. Their effects on the tumor microenvironment, cell survival, and stress adaptation position them as critical biological markers of therapeutic response. Sema3A has been shown to contribute to the development of resistance to various treatments, its inhibition in pancreatic and neuroendocrine tumor models enhanced the efficacy of the tyrosine kinase inhibitor sunitinib, leading to reduced tumor volume as well as decreased angiogenesis and metastasis.²⁰ Furthermore, Feng et al. reported that SEMA3A expression was markedly increased following chemotherapy, but gradually declined between 9 and 14 days in Lewis lung cancer rats models. Similarly, Sema7A has been shown to promote resistance mechanisms to hormonal therapies by upregulating Bcl-2 expression, contributing to tamoxifen and fulvestrant resistance. In addition, overexpression of Sema7A in lung cancer has been associated with the development of resistance to tyrosine kinase inhibitors.²¹ These data indicate that semaphorin-mediated signaling can enhance cell survival pathways, suppress apoptotic responses, and thereby reduce therapeutic efficacy.
Semaphorins have also been shown to play notable roles in the context of platinum-based chemotherapy. Sema3B was overexpressed following cisplatin treatment, which may represent a potential risk factor for cisplatin resistance.²² Additionally, targeting Sema4D was shown to enhance the chemotherapeutic response to 5-FU, suppressing both cell viability and migration in the SW48 cell line.²³
The findings of this study are consistent with the literature, indicating that platinum-based agents exert a regulatory effect on Sema5B expression. Cisplatin treatment induced a robust, time-dependent increase in Sema5B protein levels in HepG2 cells, whereas oxaliplatin elicited a more modest increase in SW480 cells. In terms of temporal dynamics, Sema5B expression in the HepG2 cell line progressively increased at 24 and 72 hours, whereas in SW480 cells, expression decreased at 72 hours relative to 24 hours. These differences may reflect cell line specific drug uptake, metabolic adaptation capacity, or biological heterogeneity in the activation of stress-response pathways. Notably, in other tumor models, semaphorin expression has also been reported to rise during the early phase following chemotherapy and subsequently decline, a pattern thought to reflect acute stress responses and adaptive reprogramming processes.
The upregulation of Sema5B following platinum treatment suggests that this protein may play a role in adaptive cellular responses to chemotherapeutic stress. Sema5B-mediated signaling could potentially contribute to treatment tolerance through mechanisms such as apoptosis suppression, activation of cell survival pathways, or cytoskeletal remodeling. In this context, Sema5B may represent not merely a passive marker of chemotherapy response but also an active regulatory candidate involved in the development of resistance.
Conversely, semaphorins are not always associated with resistance in tumor cells, and certain members have been shown to enhance chemosensitivity. For instance, Sema3F has been reported to increase chemosensitivity in colorectal cancer through p27 mediated cell cycle regulation.²⁴ In addition, tumors expressing Sema4A exhibited improved responses to immunotherapeutic agents, particularly anti-PD-1 therapy.²⁵ These contrasting effects highlight the complex and pleiotropic roles of the semaphorin family in tumor biology, which are context dependent and vary according to tumor type and treatment modality.

Limitations

This study has several limitations. First, as the findings are based solely on in vitro cell line models, they may not fully recapitulate the complexity of the tumor microenvironment or clinical conditions. The analyses were conducted in a limited number of cell lines, which restricts the generalizability of the results to tumors with different genetic backgrounds. Moreover, the study primarily focused on changes in expression levels, and detailed functional and mechanistic analyses confirming a causal role for Sema5B in chemotherapy response were not included. Although bioinformatic data support increased Sema5B expression in tumor tissues, direct correlations with clinical parameters were not established. Therefore, further translational studies incorporating in vivo models, diverse cancer subtypes, and clinical samples are warranted to clarify the role of Sema5B in chemotherapy resistance and treatment response.

Conclusion

In this study, we demonstrate that Sema5B may play a regulatory role in the cellular response to platinum-based chemotherapy and participate in the adaptive processes of cancer cells, with responses exhibiting cell type specific dynamics. This study demonstrates that Sema5B expression is modulated under chemotherapeutic stress, indicating that the molecule may serve not only as a component of tumor biology but also as a potential biomarker and therapeutic target influencing treatment response. Furthermore, our findings provide preliminary evidence that can serve as a foundation for future studies investigating the role of Sema5B in chemotherapy response. Nevertheless, more comprehensive mechanistic investigations and validation analyses are required to clarify the functional and clinical significance of this relationship.

Declarations

Abbreviations

COAD – Colon adenocarcinoma
DMSO – Dimethyl sulfoxide
DMEM – Dulbecco’s modified Eagle medium
FBS – Fetal bovine serum
HRP – Horseradish peroxidase
IC50 – Half-maximal inhibitory concentration
LIHC – Liver hepatocellular carcinoma
MTT – 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
PBS – Phosphate-buffered saline
SD – Standard deviation
Sema5B – Semaphorin 5B
TCGA – The Cancer Genome Atlas
TBS-T – Tris-buffered saline with Tween 20

References

1. Gorgulu Akin B, Erkoc M, Korkmaz ET, et al. Rapid drug desensitization with platin-based chemotherapy: analysis of risk factors for breakthrough reactions. World Allergy Organ J. 2022;15(1):100619. doi:10.1016/j.waojou.2021.100619
   Article PubMed Google Scholar
2. Ma M, He J, Gao B, et al. Targeted therapy of non-small cell lung cancer and liver cancer: functional nanocarriers for the delivery of cisplatin and tissue factor pathway inhibitor-2. Chemotherapy. 2023;68(2):73-86. doi:10.1159/000527536
   Article PubMed Google Scholar
3. Buckarma E, Thiels CA, Jin Z, Grotz TE. Cytoreduction and hyperthermic intraperitoneal paclitaxel and cisplatin for gastric cancer with peritoneal metastasis. Ann Surg Oncol. 2024;31(1):622-629. doi:10.1245/s10434-023-14379-2
   Article PubMed Google Scholar
4. Li H, Wang C, Lan L, et al. PARP1 inhibitor combined with oxaliplatin efficiently suppresses oxaliplatin resistance in gastric cancer-derived organoids via homologous recombination and the base excision repair pathway. Front Cell Dev Biol. 2021;9:719192. doi:10.3389/fcell.2021.719192
   Article PubMed Google Scholar
5. Bilgehan Sahin A, Melek H, Ocak B, et al. Platin-based chemotherapy does not improve survival in patients with nonmetastatic resected typical carcinoid tumors. Mol Clin Oncol. 2022;17(4):146. doi:10.3892/mco.2022.2579
   Article PubMed Google Scholar
6. Fernández-Nogueira P, Linzoain-Agos P, Cueto-Remacha M, et al. Role of semaphorins, neuropilins, and plexins in cancer progression. Cancer Lett. 2024;606:217308. doi:10.1016/j.canlet.2024.217308
   Article PubMed Google Scholar
7. Alto LT, Terman JR. Semaphorins and their signaling mechanisms. Methods Mol Biol. 2017;1493:1-25. doi:10.1007/978-1-4939-6448-2_1
   Article PubMed Google Scholar
8. Ding J, Zhao S, Chen X, et al. Prognostic and diagnostic values of semaphorin 5B and its correlation with tumor-infiltrating immune cells in kidney renal clear-cell carcinoma. Front Genet. 2022;13:835355. doi:10.3389/fgene.2022.835355
   Article PubMed Google Scholar
9. Li J, Hu J, Yang Y, et al. Drug resistance in cancer: molecular mechanisms and emerging treatment strategies. Mol Biomed. 2025;6(1):111. doi:10.1186/s43556-025-00352-w
   Article PubMed Google Scholar
10. Zhang C, Xu C, Gao X, Yao Q. Platinum-based drugs for cancer therapy and antitumor strategies. Theranostics. 2022;12(5):2115-2132. doi:10.7150/thno.69424
    Article PubMed Google Scholar
11. Yaron A, Huang PH, Cheng HJ, Tessier-Lavigne M. Differential requirement for Plexin-A3 and -A4 in mediating responses of sensory and sympathetic neurons to distinct class 3 semaphorins. Neuron. 2023;111(22):3697. doi:10.1016/j.neuron.2023.10.029
    Article PubMed Google Scholar
12. Worzfeld T, Offermanns S. Semaphorins and plexins as therapeutic targets. Nat Rev Drug Discov. 2014;13(8):603-621. doi:10.1038/nrd4337
    Article PubMed Google Scholar
13. Wei L, Li H, Tamagnone L, You H. Semaphorins and their receptors in hematological malignancies. Front Oncol. 2019;9:382. doi:10.3389/fonc.2019.00382
    Article PubMed Google Scholar
14. Qamar T, Misra DP, Kar S. Semaphorins and their receptors: emerging cellular biomarkers and therapeutic targets in autoimmune and inflammatory disorders. Life Sci. 2025;361:123281. doi:10.1016/j.lfs.2024.123281
    Article PubMed Google Scholar
15. Chen Y, Zhang L, Lv R, Zhang WQ. Overexpression of semaphorin 4D indicates poor prognosis and prompts monocyte differentiation toward M2 macrophages in epithelial ovarian cancer. Asian Pac J Cancer Prev. 2013;14(10):5883-5890. doi:10.7314/apjcp.2013.14.10.5883
    Article PubMed Google Scholar
16. Ruan SS, Li RC, Han Q, et al. Expression and clinical significance of semaphorin 4D in non-small cell lung cancer and its impact on malignant behaviors of A549 lung cancer cells. J Huazhong Univ Sci Technolog Med Sci. 2014;34(4):491-496. doi:10.1007/s11596-014-1304-2
    Article PubMed Google Scholar
17. Lee J, Shin YJ, Lee K, et al. Anti-SEMA3A antibody: a novel therapeutic agent to suppress glioblastoma tumor growth. Cancer Res Treat. 2018;50(3):1009-1022. doi:10.4143/crt.2017.315
    Article PubMed Google Scholar
18. Zhang C, Zhang L, Zhang X, et al. SEMA5B depletion suppresses cell proliferation and glycolysis by downregulating VDAC2 expression in esophageal squamous cell carcinoma. Int J Biol Macromol. 2026;341(pt 2):149690. doi:10.1016/j.ijbiomac.2025.149690
    Article PubMed Google Scholar
19. Doughan A, Salifu SP. Genes associated with diagnosis and prognosis of Burkitt lymphoma. IET Syst Biol. 2022;16(6):220-229. doi:10.1049/syb2.12054
    Article PubMed Google Scholar
20. Maione F, Capano S, Regano D, et al. Semaphorin 3A overcomes cancer hypoxia and metastatic dissemination induced by antiangiogenic treatment in mice. J Clin Invest. 2012;122(5):1832-1848. doi:10.1172/jci58976
    Article PubMed Google Scholar
21. Kinehara Y, Nagatomo I, Koyama S, et al. Semaphorin 7A promotes EGFR-TKI resistance in EGFR mutant lung adenocarcinoma cells. JCI Insight. 2018;3(24). doi:10.1172/jci.insight.123093
    Article PubMed Google Scholar
22. Peszek W, Kras P, Grabarek BO, Boron D, Oplawski M. Cisplatin changes expression of SEMA3B in endometrial cancer. Curr Pharm Biotechnol. 2020;21(13):1368-1376. doi:10.2174/1389201021666200514215839
    Article PubMed Google Scholar
23. Rashidi G, Rezaeepoor M, Mohammadi C, Solgi G, Najafi R. Inhibition of semaphorin 4D enhances chemosensitivity by increasing 5-fluorouracil-induced apoptosis in colorectal cancer cells. Mol Biol Rep. 2020;47(9):7017-7027. doi:10.1007/s11033-020-05761-4
    Article PubMed Google Scholar
24. Tao M, Ma H, Fu X, et al. Semaphorin 3F induces colorectal cancer cell chemosensitivity by promoting P27 nuclear export. Front Oncol. 2022;12:899927. doi:10.3389/fonc.2022.899927
    Article PubMed Google Scholar
25. Naito Y, Koyama S, Masuhiro K, et al. Tumor-derived semaphorin 4A improves PD-1-blocking antibody efficacy by enhancing CD8+ T-cell cytotoxicity and proliferation. Sci Adv. 2023;9(20). doi:10.1126/sciadv.ade0718
    Article PubMed Google Scholar

Additional Information

Publisher’s Note
Bayrakol MP remains neutral with regard to jurisdictional and institutional claims.

Rights and Permissions

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0). To view a copy of the license, visit https://creativecommons.org/licenses/by-nc/4.0/

View full license details →

About This Article

Received:

March 3, 2026

Published Online:

June 22, 2026