Postexercise downregulation of NUP155 in regulating non-small cell lung cancer progression via the PTEN/AKT signaling pathway

Introduction

Lung cancer (LC) continues to be the leading cause of cancer-related death worldwide, with non-small cell lung cancer (NSCLC) being particularly linked to well-established risk factors such as smoking, radon gas exposure, asbestos exposure, environmental contaminant exposure, and genetic predispositions (1,2). Despite advancements in treatment options such as surgery, chemotherapy, radiotherapy, and targeted therapies, the prognosis of many patients with LC remains poor, highlighting the need for novel diagnostic biomarkers, therapeutic strategies, and prognostic indicators to improve patient outcomes (3). Recent studies have begun to explore the potential relationship between physical activity and the progression of NSCLC, suggesting that exercise may play a crucial role in disease management (4,5). Research indicates that physical activity may enhance overall survival (OS) rates and quality of life for patients with LC by improving immune function, reducing systemic inflammation, and enhancing physical fitness (6). These results imply that exercise may also reduce the risk of cancer recurrence and enhance tolerance to treatments such as chemotherapy and radiation therapy. Additionally, physical activity can mitigate treatment-related side effects, thereby improving patients’ functional capacity and mental health (7). Despite these promising outcomes, the exact mechanisms by which exercise influences LC progression are not yet fully understood. Ongoing studies are investigating various pathways and biological processes that are affected by exercise, including immune response modulation, hormonal changes, and alterations in the tumor microenvironment (8,9). Among these pathways, the PI3K/AKT signaling pathway, regulated by the PTEN tumor suppressor, is essential to cell survival and proliferation, making it a key area of interest in cancer research (10). Given the significant burden and high mortality rate of LC, exploring exercise as an adjunct therapy may hold considerable clinical relevance.

NUP155 encodes a key protein component of the nuclear pore complex (NPC), which is necessary for regulating the movement of molecules from the cytoplasm to the nucleus (11). Recent research has highlighted a possible connection between cancer and NUP155. Disruptions in NUP155 expression or function can lead to aberrant nucleocytoplasmic transport, resulting in altered cell cycle regulation and genomic instability hallmarks of cancer (12). For instance, research by Forest et al. revealed that NUP155 expression is significantly higher in grade 3 lung adenocarcinoma (LUAD) compared to grade 2, suggesting its involvement in disease progression and severity, as well as its potential role in chemoresistance (13). Furthermore, a pan-cancer analysis by Wang et al. identified NUP155 as an important contributor to the emergence and spread of tumors (14). The study noted NUP155 upregulation in 26 different cancer types, with associations to advanced pathological stages and poor prognosis. Microsatellite instability, stemness score, tumor mutational load, and DNA methylation correlate with NUP155 expression, implicating it in genomic instability and cancer stem cell characteristics. Additionally, NUP155 influences the tumor immune microenvironment (TIME), affecting immune cells that invade tumors and the genes linked to immunoregulation (15). Moreover, the TIME is involved in immunomodulatory pathways, particularly in antigen processing and presentation. In breast cancer, NUP155 promotes cell proliferation, migration, and resistance to apoptosis. Savci-Heijink et al. identified 15 signature genes, including NUP155, that were predictive of bone metastasis in breast carcinomas (16). This signature significantly correlated with metastatic behavior and survival outcomes, emphasizing the role of NUP155 in the metastatic process and highlighting its potential as a therapeutic target and prognostic indicator for different types of cancer.

The interplay between physical activity and cancer biology has garnered increasing attention, with a study suggesting that exercise can influence gene expression and tumor progression (17). NUP155, a nucleoporin gene, has been implicated in various malignancies, including its association with LUAD (13,14). However, the relationship between exercise-related modulation of NUP155 expression and its potential impact on cancer prognosis remains unexplored. The objective of this study was thus to examine the expression levels of NUP155 in NSCLC and assess its role in the PTEN/AKT signaling pathway, particularly in the context of physical activity. By elucidating the mechanisms through which NUP155 affects NSCLC cell behavior, we aimed to gain a novel understanding of the molecular causes of cancer progression and explore the potential therapeutic strategies that integrate lifestyle interventions, such as exercise, for improved cancer management. We present this article in accordance with the MDAR reporting checklist (available at https://tcr.amegroups.com/article/view/10.21037/tcr-24-1619/rc).

Methods

Ethical statement

The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013).

Differential expression analysis of relevant datasets

The GSE41914 dataset was obtained from the Gene Expression Omnibus (GEO) (accession no. GSE41914; https://www.ncbi.nlm.nih.gov/geo/) and comprises 12 samples of natural killer (NK) cells collected both before and after exercise. Additionally, 1,017 NSCLC samples and 108 control normal samples were downloaded from the Clinical Bioinformatics website (https://www.aclbi.com/static/index.html#/). These two datasets were employed to determine differentially expressed genes (DEGs) via the “Limma” package in version 3.42.2 (The R Foundation for Statistical Computing). DEGs were classified based on fold change (FC) values: genes with an FC greater than 1.3 were considered upregulated, while those with an FC less than 0.77 were considered downregulated. A P value <0.05 indicated that genes were both upregulated and downregulated.

Expression and prognostic analysis of NUP155

The Venn online graph tool (https://bioinformatics.psb.ugent.be/webtools/Venn/) was used to filter upregulated DEGs and downregulated DEGs in the GSE41914 dataset and the Cancer Genome Atlas (TCGA)-NSCLC dataset. We selected NUP155 among the overlapping genes as a hub gene for further analysis. The expression levels of NUP155 in the GSE41914 dataset and the TCGA-NSCLC dataset were analyzed via the Sangerbox 3.0 website (http://vip.sangerbox.com/home.html). Subsequently, the cohorts in the GSE41914 dataset and the TCGA-NSCLC dataset were divided into two risk categories (low risk and high risk) according to the expression patterns of the relevant genes. The Wilcoxon test was used to assess the variations in NUP155 expression levels across the groups. The Kaplan-Meier (KM) analysis was employed to determine the OS and first progression probabilities (FPPs) of the two risk groups. In addition, the statistical significance of the survival difference between the two groups was assessed using the log-rank test. The hazard ratio (HR) of the high-risk group was also calculated to further clarify the relative risk.

Cell lines and culture

The National Collection of Authenticated Cell Cultures (Shanghai, China) provided human normal lung cells (BEAS-2B; cat. No. SCSP-5067) and NSCLC cells (A549, cat. No. SCSP-503; H1299, cat. No., SCSP-589; H1975, cat. No. SCSP-597), while the BeNa Culture Collection (Beijing, China) provided another NSCLC cell line, H157 (cat. No. BNCC359887). Their diet consisted of Dulbecco’s Modified Eagle Medium (DMEM) with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 2 mM of L-glutamine added as supplements. Cell cultures were maintained in an enhanced humidity environment with 5% CO₂ at 37 ℃. Mycoplasma contamination testing was conducted regularly, with negative results confirmed before each experimental setup. Cell lines were used at passages below 20 to minimize the risk of phenotypic drift, while primary cultures were used within the first 5 passages to preserve their native cellular characteristics. No genetic modifications were introduced to the primary cultures during the experiments.

Cell treatment and transfection

SF1670 is a small molecule inhibitor specifically designed to inhibit PTEN activity (18). NSCLC cells received treatment of 10 µM of SF1670 for 24 hours. For transient transfection, NSCLC cells were transfected with two distinct small interfering RNAs (siRNAs) that target NUP155 to attain knockdown of NUP155 expression, and cells were cultured for a certain amount of time to effectively knockdown NUP155. Cells were transfected using Lipofectamine 3000 (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions.

Quantitative real-time polymerase chain reaction (qRT-PCR)

According to a previous study, qRT-PCR was performed (19). The data were examined using the 2^-ΔΔCT approach, with GAPDH abundance serving as the standard. The amplification procedure made use of the primer sequences listed below: NUP155 forward, 5'-CTTAGTGTCTACCTGGCTGCTTGG-3'; and NUP155 reverse, 5'-TGATGCTGATGCTGATGCTTCTGG-3'. Similarly, the forward and reverse primers for GAPDH, which was used as the reference gene, were as follows: GAPDH forward, 5'-ATTCCACCCATGGCAAATT-3'; and GAPDH reverse, 5'-TGGGATTTCCATTGATGACAAG-3'.

Western blotting

Protease and phosphatase inhibitors (CoWin Biosciences, Nanjing, China) were added to a RIPA lysis buffer (Solarbio, Beijing, China) to facilitate the preparation of protein lysates from NSCLC cells. The protein content was determined using the BCA Protein Assay Kit (Beyotime, Shanghai, China). Proteins in equal quantities were divided using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then placed onto polyvinylidene fluoride membranes (Beyotime). The membranes were blocked with 5% skim milk and incubated with the following primary antibodies (each at a 1:1,000 dilution): NUP155 (cat. No. ab199528; Abcam, Cambridge, UK), Bax (cat. No. ab32503; Abcam), Bcl-2 (cat. No. ab32124; Abcam), caspase-9 (cat. No. ab32539, Abcam), PTEN (cat. No. ab267787; Abcam), p-AKT (cat. No. ab8933; Abcam), and AKT (cat. No. ab192623; Abcam). After a wash, the membranes were incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG) secondary antibody (cat. No. ab6721; Abcam) at a 1:5,000 dilution for 1 hour at room temperature. GAPDH (1:5,000; cat. No. KC-5G5; Kangcheng, Shanghai, China) was included for internal use only. An enhanced chemiluminescence (ECL) kit (Tiangen, Beijing, China) was used to visualize the protein bands, and ChemiDoc imaging equipment (Bio-Rad, Hercules, CA, USA) was used to obtain the images.

Cell Counting Kit 8 (CCK8) assay

CCK8 (KeyGEN, Nanjing, China) was used to determine the viability of the cells. NSCLC cells were planted in 96-well plates at a concentration of 5×10³ per well. After a CCK8 reagent was added to each well after treatment, a microplate reader (Kehua Technologies, Inc., Shanghai, China) was used to measure the absorbance at 450 nanometers after 0, 24, 48, 72, 96, and 120 hours.

Flow cytometry

NSCLC cells were separated using 0.25% trypsin-EDTA (Life Technologies Inc., Carlsbad, CA, USA) and then cleaned with phosphate-buffered saline (PBS) in preparation for flow cytometry. To distinguish viable, apoptotic, and necrotic cells, the annexin V and propidium iodide (PI) staining procedures were completed in line with the manufacturer’s instructions. A flow cytometer (Jiyuan Biotechnology Co., Ltd., Guangzhou, China) was used for the flow cytometry, and FlowJo software (FlowJo, BD Biosciences, Franklin Lakes, NJ, USA) was used for data analysis to calculate the cell apoptosis rate.

Cell invasion and migration assays

Transwell assay was used to measure migration and invasion of cells. Transfected NSCLC cells were suspended in serum-free media within the top chamber of the Transwell (Corning, Corning, NY, USA). Subsequently, 10% FBS was included in the medium in the lower chamber of the Transwell. After an incubation period, cells with moving cell membranes were stained with DAPI and fixed with 4% paraformaldehyde. Finally, to count the number of migrating cells in the field of vision, inverted microscopy was used. The aforementioned cell invasion assays were performed with Matrigel (BD Biosciences, Shanghai, China) covering the top chamber.

Statistical analysis

R was used to carry out statistical analyses. Each experiment was performed with three technical replicates, and the mean ± standard deviation (SD) of the findings is given. In order to assess the importance of the variations, one-way analysis of variance was used, and the Tukey test was used for post hoc analysis. P<0.05 was established as the threshold for statistical significance.

Results

A high expression of exercise-related gene NUP155 in NSCLC was associated with poor prognosis

A total of 589 upregulated DEGs and 299 downregulated DEGs were identified between pre-exercise and postexercise samples from the GSE41914 dataset, alongside 4,571 upregulated DEGs and 4,223 downregulated DEGs between the TCGA-NSCLC dataset’s tumor and normal samples (Figure 1A,1B). Additionally, 204 overlapping genes were identified from the upregulated and downregulated DEGs of the GSE41914 and TCGA-NSCLC datasets (Figure 1C). Notably, NUP155 was selected as a hub gene for further analysis. In the GSE41914 dataset, NUP155 expression levels were noticeably lower in postexercise samples than in pre-exercise samples, suggesting that this gene may be regulated by physical activity (Figure 1D). Furthermore, in TCGA dataset, NUP155 expression was noticeably elevated in NSCLC tissues in contrast to normal tissues (Figure 1E). High NUP155 expression was linked to poor OS and FPP in patients with NSCLC according to the KM survival analysis (Figure 1F,1G). These findings imply that NUP155 could function as an NSCLC prognostic biomarker and that its elevated expression is associated with poor clinical outcomes.

Figure 1 Differential expression and prognostic significance of NUP155 in NSCLC-related datasets. (A,B) DEGs screening of the GSE41914 dataset and TCGA-NSCLC dataset. Blue represents the downregulated DEGs, and red represents the upregulated DEGs. (C) Cross-analysis of upregulated and downregulated DEGs in the GSE41914 and TCGA-NSCLC datasets for obtaining the overlapping genes. (D) Boxplot showing the expression level of NUP155 in pre-exercise and postexercise samples in the GSE41914 dataset. (E) Boxplot showing the expression level of NUP155 in NSCLC samples and normal samples in TCGA dataset. (F) Kaplan-Meier survival curve showing the patients’ overall chance of survival from NSCLC. The red line represents high NUP155 expression, and the blue line represents low NUP155 expression. (G) Kaplan-Meier survival curve of first progression probability of patients with NSCLC. The red line represents high NUP155 expression, and the blue line represents low NUP155 expression. **, P<0.01; ****, P<0.0001. NSCLC, non-small cell lung cancer; DEG, differentially expressed gene; TCGA, The Cancer Genome Atlas.

NUP155 was upregulated in NSCLC cells

In NSCLC cell lines (A549, H157, H1299, and H1975), the expression of NUP155 was assessed and compared with that of normal lung cells (BEAS-2B). NUP155 messenger RNA (mRNA) and protein levels were revealed to be significantly elevated in A549, H1299, and H1975 cells, particularly in A549 and H1299, via qRT-PCR and Western blotting (Figure 2A-2C). Through the use of specific siRNAs (si-NUP155-1 and si-NUP155-2), NUP155 expression was effectively knocked down in H1299 and A549 cells, as shown by the reduced amounts of mRNA and proteins (Figure 2D-2F). These results suggest that NUP155 is overexpressed in NSCLC and could serve as a possible therapeutic target.

Figure 2 NUP155 was upregulated in NSCLC cells. (A) qRT-PCR measurements of the relative mRNA expression of NUP155 in NSCLC cell lines (A549, H157, H1299, and H1975) and normal lung cells (BEAS-2B). (B,C) WB of NUP155 protein levels in BEAS-2B and NSCLC cell lines. (D) Relative mRNA expression of NUP155 in A549 and H1299 cells after transfection with control siRNA (si-NC) or two different NUP155-targeting siRNAs (si-NUP155-1 and si-NUP155-2). (E,F) WB of NUP155 protein knockdown efficiency in A549 and H1299 cells. *, P<0.05; **, P<0.01. qRT-PCR, quantitative real-time polymerase chain reaction; NSCLC, non-small cell lung cancer; WB, western blotting; si-NC, small interfering RNA negative control; siRNA, small interfering RNA.

Knockdown of NUP155 inhibited the proliferation, migration, and invasion of NSCLC cells

The CCK8 assay demonstrated a significant reduction in NSCLC cell viability following NUP155 knockdown, with si-NUP155-2 exhibiting a more pronounced inhibitory effect (Figure 3A,3B). Similarly, transwell assay indicated that NUP155 knockdown notably decreased the invasion and migration abilities of NSCLC cells after 72 hours, with si-NUP155-2 showing a greater reduction (Figure 3C-3F). These findings suggest that NUP155 knockdown can affect NSCLC cell phenotypes, indicating its potential as a therapeutic target for inhibiting tumor progression.

Figure 3 Knockdown of NUP155 inhibited the proliferation, migration, and invasion of NSCLC cells. (A,B) CCK8 assay of NSCLC cells (A549 and H1299) at various time points (0, 24, 48, 72, 96, and 120 hours) following NUP155 knockdown. (C-F) Transwell assay for detecting the changes in the invasion and migration ability of NSCLC cells (A549 and H1299) after knockdown of NUP155. Cells were stained using DAPI and imaged with a fluorescence microscope. Scale bar: 50 µm. *, P<0.05; **, P<0.01; ***, P<0.001. OD, optical density; NSCLC, non-small cell lung cancer; CCK8, Cell Counting Kit 8.

Knockdown of NUP155 promoted the apoptosis of NSCLC cells

Flow cytometry with annexin V and PI staining demonstrated that NUP155 knockdown significantly increased the apoptotic rate of A549 and H1299 cells as compared with control siRNA (si-NC) (Figure 4A-4C). After treatment with si-NUP155-1 and si-NUP155-2, the percentage of dying cells in both cell lines was significantly higher, indicating effective induction of apoptosis. Analysis at the protein level further confirmed these findings, showing that after NUP155 knockdown in A549 and H1299 cells, the expression of the proapoptotic proteins Bax and caspase-9 was elevated, whereas the expression of the anti-apoptotic protein Bcl-2 was decreased (Figure 4D-4F). These findings suggest that NUP155 knockdown promotes apoptosis in NSCLC cells by regulating the expression of key apoptotic regulators, highlighting its potential as a target for NSCLC treatment intervention.

Figure 4 Knockdown of NUP155 promoted apoptosis of NSCLC cells. (A-C) Following NUP155 knockdown, apoptosis in NSCLC cells (A549 and H1299) was observed via flow cytometry. Quadrants represent viable cells (Q4), late apoptotic cells (Q2), early apoptotic cells (Q3), and necrotic cells (Q1). (D-F) WB was used to detect the protein expression of apoptotic proteins in NSCLC cells (A549 and H1299) after the knockdown of NUP155. *, P<0.05; **, P<0.01; ***, P<0.001. NSCLC, non-small cell lung cancer; WB, western blotting.

NUP155 activated the PTEN/AKT signaling pathway in NSCLC cells and influenced tumor migration and invasion

PTEN is a critical tumor suppressor that inhibits the PI3K/AKT signaling pathway by dephosphorylating PIP3 (20). The phosphorylated form of AKT (p-AKT) serves as a marker that initiates this pathway (21). Western blotting revealed that the knockdown of NUP155 in NSCLC cells significantly upregulated PTEN protein levels while markedly downregulating p-AKT levels, with no significant change in total AKT levels (Figure 5A-5C). This suggests that NUP155 may regulate the PTEN/AKT signaling pathway, influencing cancer cell behavior. To gain further insight into the function of PTEN in this context, we treated NSCLC cells with SF1670, a small-molecule inhibitor specifically targeting PTEN activity, for 24 hours. Transwell assays showed that the addition of SF1670 alleviated the reduced migration and invasion capabilities induced by NUP155 knockdown (Figure 5D-5G). These results point to the involvement of NUP155 in regulating the PTEN/AKT signaling pathway, with subsequent effects on NSCLC cell invasion and migration.

Figure 5 NUP155 activated the PTEN/AKT signaling pathway in NSCLC cells and influenced tumor migration and invasion. (A-C) WB detection of PTEN, p-AKT, and AKT protein levels in NSCLC cells after knockdown of NUP155. *, P<0.05; **, P<0.01; ***, P<0.001. (D-G) Transwell detection of the invasion and migration ability of NSCLC cells after knockdown of NUP155 and addition of PTEN inhibitor SF1670. Cells were stained using DAPI and imaged with a fluorescence microscope. Scale bar: 50 µm. **, P<0.01; ***, P<0.001 vs. si-NC. ^#, P<0.05; ^##, P<0.01 vs. si-NUP155-2. WB, western blotting; NSCLC, non-small cell lung cancer; si-NC, small interfering RNA negative control.

Discussion

Physical activity has well-documented comprehensive health benefits, including cancer prevention and management. Regular exercise has been shown to lower the risk of developing various cancers, such as breast, colorectal, and LCs (22). This protective effect is partly attributed to exercise’s ability to enhance immune function, reduce systemic inflammation, and improve metabolic regulation (23). For example, exercise can lower levels of insulin and insulin-like growth factors, which are known to promote cancer cell proliferation (24). Additionally, physical activity can modulate hormone levels, decrease obesity, and improve cardiovascular health, all of which contribute to a reduced cancer risk (25). In the context of LC, physical activity has been associated with improved treatment outcomes and quality of life, suggesting a potential role in modulating disease progression (26,27). Our study adds to this body of evidence by exploring the relationship between physical activity and the expression of exercise-related genes, specifically NUP155, which may further clarify the underlying molecular processes underlying the benefits of exercise in cancer prevention and management.

NUP155, a nucleoporin gene, plays a crucial role in regulating the NPC and cellular transport mechanisms (28). Our study identified NUP155 as a key exercise-related gene, exhibiting significant downregulation in post-exercise samples and upregulation in NSCLC samples, which was associated with poorer prognosis and tumor progression through enhanced proliferation, invasion, and migration in NSCLC cells. The results are consistent with findings from Holzer et al., who demonstrated that NUP155 regulates the mRNA translation of p21, influencing cancer progression via translational control mechanisms involving FTSJ1 and p53 status (29). Additionally, Chi et al. highlighted the function of NUP155 in cell proliferation and survival through RNA export and interaction with other nucleoporins, which is linked to a poor prognosis and an advanced stage of the illness (30). Our study further showed that knockdown of NUP155 significantly increased apoptosis in NSCLC cells, as seen by the downregulation of the antiapoptotic protein Bcl-2 and the overexpression of the proapoptotic proteins Bax and caspase-9. Apoptosis, which is essential for tissue homeostasis, involves key regulators such as Bax and Bcl-2, which govern mitochondrial membrane permeability and caspase activation (31). In NSCLC, apoptosis is the key to effective treatment. Zuo et al. found that IFN-α and celecoxib synergistically enhanced TRAIL-induced apoptosis, and Zu et al. found that miR-146a functions to regulate apoptosis and cell proliferation (32,33). Similarly, Zuo et al. reported that bamboo saponins IV and V induced apoptosis through a p53-mediated pathway (34). Our results indicate that NUP155 figures prominently in NSCLC progression and apoptosis, highlighting its potential use as a target for therapy in the context of physical activity and cancer management.

The PTEN signaling pathway is crucial in regulating cell growth, survival, and proliferation, and serves as a suppressor of tumors by inhibiting the PI3K/AKT signaling pathway (35,36). In cancers, including NSCLC, mutation or loss of PTEN function leads to uncontrolled cell growth and tumor development (37,38). PTEN inhibits AKT activation by dephosphorylating PIP3 to PIP2, thereby preventing downstream signaling that promotes cell survival and proliferation (39). Consequently, PTEN dysfunction is often associated with poor prognosis and resistance to therapies. For example, Zou et al. found that KBTBD7 promotes the ubiquitin-dependent degradation of PTEN, resulting in the engagement of the EGFR/PI3K/AKT pathway and promotion of NSCLC cell proliferation and invasion (40). In a similar vein, Zhu et al. demonstrated that USP52 stabilizes PTEN, downregulates cyclin D1, and inhibits the AKT/mTOR pathway, thereby suppressing cancer cell proliferation (41). In the study by Zhu et al., miR-205 was significantly upregulated in LC tissues and decreased PTEN expression and activated the PI3K/AKT pathway, thus promoting cell proliferation, migration, and invasion (42). In our study, we found that knocking down NUP155 in NSCLC cells significantly reduced p-AKT levels, elevated PTEN protein levels, and left AKT levels unchanged. Transwell assays showed that the PTEN inhibitor SF1670 mitigated the reduction in cell migration and invasion caused by NUP155 knockdown, indicating that NUP155 may modulate tumor migration and invasion via the PTEN/AKT signaling pathway. These findings highlight the importance of targeting the PTEN/AKT signaling pathway as a form of cancer therapy, particularly in terms of restoring normal cell function and inhibiting tumor progression.

NUP155 is upregulated in NSCLC and is associated with enhanced tumor proliferation, migration, and invasion. The downregulation of NUP155 after exercise may have significant implications for the progression of NSCLC. Firstly, the downregulation of NUP155 can inhibit the proliferation, migration, and invasion of NSCLC cells and promote apoptosis. This finding suggests that exercise might slow down the growth and spread of tumor cells by reducing the expression levels of NUP155, thereby exerting an inhibitory effect on the progression of NSCLC. Secondly, NUP155 influences cell invasion and migration through the PTEN/AKT signaling pathway. The downregulation of NUP155 after exercise may affect this pathway, thereby inhibiting the invasive and migratory capabilities of tumor cells, which is crucial for controlling local tumor spread and distant metastasis. Lastly, as a potential therapeutic target, the downregulation of NUP155 after exercise provides possibilities for developing new treatment strategies. Exercise, as a non-pharmacological treatment method, may enhance the effectiveness of traditional therapeutic approaches, such as chemotherapy and radiation therapy, by downregulating NUP155. It may even be combined with immunotherapy to improve treatment outcomes. In summary, the downregulation of NUP155 after exercise may affect the progression of NSCLC through multiple mechanisms, including inhibiting tumor cell proliferation and invasion, and serving as a potential therapeutic target. These findings provide new perspectives and potential treatment strategies for the treatment of NSCLC.

In our study, we utilized bioinformatics analysis to assess the expression levels of NUP155 both before and after exercise, and we also compared these levels between NSCLC samples and normal tissue samples. When analyzing samples before and after exercise, we used NK cell data, however, this has certain limitations. Due to different cellular mechanisms, complexity of tumor microenvironment, and lack of direct cellular response data, the response of NK cells to motility does not fully represent the direct impact on NSCLC. The interaction between NK cells and NSCLC cells is complex, and changes in NK cell function may not be a direct indicator of NSCLC cells' response to movement. Our in vitro experiments focused on the functional role of NUP155 in NSCLC, rather than the direct post-exercise effects, due to current experimental constraints. To overcome these limitations, future research will include direct measurement of post-exercise NUP155 levels in NSCLC cell lines to elucidate the immediate impact of exercise on NUP155 expression. Additionally, we will explore the effects of exercise on NUP155 expression in preclinical models, such as xenografts, to bridge the gap between in vitro and in vivo studies and gain a more holistic view of exercise’s influence on NUP155 expression and function in a physiological setting.

Conclusions

Our study highlights the crucial role of the exercise-related gene NUP155 in promoting NSCLC cell proliferation, migration, invasion, and apoptosis regulation. Elevated NUP155 expression is linked to comparatively poor clinical results, suggesting its potential as a prognostic biomarker. Notably, NUP155 appears to activate the PTEN/AKT signaling pathway, influencing tumor cell behavior. Knockdown of NUP155 leads to increased PTEN expression and decreased p-AKT levels, resulting in reduced cell migration and invasion. The use of the PTEN inhibitor SF1670 further confirmed the involvement of this pathway. These findings suggest that targeting NUP155 and modulating the PTEN/AKT pathway could be a promising therapeutic strategy for NSCLC, paving the way for future research on the combination of molecular-targeted therapies and physical activity for improving patient outcomes. However, we still have an incomplete understanding as to how exercise specifically regulates NUP155 expression and lack clinical data to support its practical applications; thus warranting further investigation.

Acknowledgments

Funding: This study was funded by the Research on Adolescent Mental and Brain Health Screening, Assessment, and Intervention (No. L110-72301).

Footnote

Reporting Checklist: The authors have completed the MDAR reporting checklist. Available at https://tcr.amegroups.com/article/view/10.21037/tcr-24-1619/rc

Data Sharing Statement: Available at https://tcr.amegroups.com/article/view/10.21037/tcr-24-1619/dss

Peer Review File: Available at https://tcr.amegroups.com/article/view/10.21037/tcr-24-1619/prf

Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tcr.amegroups.com/article/view/10.21037/tcr-24-1619/coif). The authors have no conflicts of interest to declare.

Ethical Statement: The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013).

Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0/.

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(English Language Editor: J. Gray)