Aggressive invasion and metastatic dissemination of gastric cancer (GC) are two major clinical challenges that frequently arise following standard treatments, markedly compromising patient outcomes. Elucidating the molecular drivers of GC progression would be key to developing effective therapeutic strategies. The present study employed an integrated approach combining bioinformatics analysis and immunohistochemical (IHC) validation to identify the key molecular players in GC metastasis. The Cancer Genome Atlas (TCGA) database analysis demonstrated marked upregulation of both epidermal growth factor receptor (EGFR) and matrix metalloproteinase 7 (MMP7) in gastric adenocarcinoma and elevated expression levels notably associated with poor patient prognosis. MMP7 expression exhibited a particularly robust association with metastatic progression, highlighting its potential role in facilitating tumor dissemination and experimental validation using IHC analysis of clinical specimens confirmed the coordinated involvement of both phosphorylated (p)-EGFR and MMP7 in metastatic processes. Notably, the present study identified a positive correlation between p-EGFR and MMP7 expression, suggesting a potential mechanistic interplay between these molecules in driving GC metastasis. These findings provide notable evidence that p-EGFR and MMP7 collectively contribute to GC progression and metastasis. The correlation between these markers offered novel insights into potential cooperative signaling pathways and presented a rational basis for the development of dual-targeted therapeutic approaches. The present study established a key foundation for future research aimed at disrupting the metastatic pathways in GC through targeted inhibition of p-EGFR and MMP7.
phosphorylated-EGFRMMP7gastric cancermetastasisThe Hefei Medical Research ProjectHwk2023zc015The National Natural Science Foundation of China81972266The present study was supported by The Hefei Medical Research Project (grant no. Hwk2023zc015) and The National Natural Science Foundation of China (grant no. 81972266).Introduction
Gastric cancer (GC) is the fifth most common malignancy and the fourth leading cause of cancer-related mortality worldwide, accounting for ~800,000 mortalities annually (1,2). Asia has the highest global burden, with 820,000 novel cases and 576,000 mortalities reported in 2020 alone (3). Due to non-specific early symptoms (such as epigastric discomfort or dull pain, loss of appetite, early satiety, belching, acid reflux and nausea) (4,5), delayed clinical presentation and limited sensitivity of current tumor markers such as CD101 and Tim3 (6,7), the majority of patients with GC are diagnosed at advanced stages, precluding curative surgical resection. Current treatment strategies, including chemotherapy, targeted therapy and combination regimens (for example, 5-fluorouracil-based, platinum-based and newer drug combinations), have markedly improved over the past four decades. Nevertheless, persistent challenges such as tumor invasion, metastasis and recurrence often lead to treatment failure, markedly impairing patient survival and quality of life (8–10). Therefore, elucidating the molecular mechanisms underlying GC invasion and metastasis remains a key research priority to improve therapeutic outcomes in the future.
Increasing evidence indicates that dysregulated activation of key signaling pathways, including epithelial-mesenchymal transition (EMT), PI3K/AKT/mTOR, Ras/Raf/ERK, Janus kinase/STAT and epidermal growth factor receptor (EGFR)-mediated signaling, serves a key role in promoting malignant progression (11–14). The aberrantly activated pathways facilitate tumor aggressiveness by modulating downstream effector molecules. EGFR is a transmembrane tyrosine kinase receptor that is extensively expressed in mammalian epithelial cells, fibroblasts, glial cells and keratinocytes. As a key regulator of cell proliferation, survival and migration, the EGFR pathway is implicated in the pathogenesis of multiple cancer types. EGFR upregulation has been validated as a prognostic biomarker in breast cancer progression (15). Previous studies have reported that EGFR can be used as a marker in predicting breast cancer progression and prognosis (16–18). Furthermore, EGFR has been reported to be associated with the metastasis of gallbladder, bladder, lung and colon cancer (19–22). Notably, emerging studies have associated EGFR dysregulation with the invasion and metastasis of GC, highlighting its potential as a therapeutic target (23,24).
Postoperative recurrence, invasion and metastasis are major challenges in GC management that markedly compromise patient survival and quality of life (25,26). Among the molecular mediators of these processes, matrix metalloproteinases (MMPs) serve a well-established role in facilitating GC progression (27,28). MMP7, a key member of the MMP family, exhibits unique characteristics despite its relatively simple structure. With a molecular weight of only 19 kDa upon activation, MMP7 demonstrates notably broad substrate specificity, degrading both extracellular matrix (ECM) and non-ECM components to promote tumor invasion and metastasis. Emerging evidence indicates that MMP7 is upregulated in multiple malignancies, including GC, hepatocellular carcinoma and colorectal cancer (29–31). Notably, MMP7 differs from other MMP family members in its tumor cell-specific secretion pattern (vs. stromal cell-derived production), making it a potential diagnostic biomarker as well as a promising therapeutic target (32). The aforementioned findings underscore the key involvement of MMP7 in GC recurrence, invasion and metastasis.
Although the individual roles of EGFR and MMP7 in cancer are well-recognized, their direct association and functional interdependence in GC remain insufficiently explored. Phosphorylation of EGFR at tyrosine 1068 (p-EGFR) is a well-established indicator of its activation and downstream signaling, with significant implications in tumor progression (33–35). To clarify the clinical relevance of this modification in the context of MMP7 co-expression, p-EGFR was specifically assessed in the present study. The present study integrated The Cancer Genome Atlas (TCGA) data with clinical immunohistochemical (IHC) validation to assess their co-expression as a hallmark of metastatic GC. Through comprehensive analysis of the TCGA database, the expression profiles of EGFR and MMP7 and their association with clinical outcomes in patients with GC were examined. Furthermore, IHC analysis of clinical GC specimens was conducted to explore the correlation between p-EGFR and MMP7 expression. Collectively, the present study aimed to establish whether p-EGFR and MMP7 function as key mediators in GC metastasis, thereby providing insights that could inform future therapeutic strategies targeting GC invasion and metastasis.
Materials and methodsTissue samples
This was a retrospective cohort study. GC tissue samples (n=32) and their corresponding normal tissue samples (samples taken at a distance of ≥2.5 cm from the cancer tissue) were obtained from patients with GC who underwent surgery at The First Affiliated Hospital of Anhui Medical University (Hefei, China) from November 2022 to December 2023. Tumor classification was based on the World Health Organization (WHO) Classification of Tumors, 5th edition (36). The inclusion criteria were as follows: i) Underwent a definitive surgical resection for primary GC between November 2022 to December 2023; ii) had a confirmed histopathological diagnosis of GC according to the WHO Classification of Tumors, 5th edition; iii) formalin-fixed, paraffin-embedded tumor tissue blocks with sufficient quality and quantity were available for subsequent molecular and IHC analyses; and iv) had complete clinicopathological data and follow-up records that were accessible from the institutional database. The exclusion criteria were as follows: i) Received any form of neoadjuvant chemotherapy or radiotherapy prior to surgical resection; ii) had a history of other synchronous or metachronous active malignancies within 5 years prior to the diagnosis of the index tumor; iii) presented with distant metastasis (Stage 4 disease) at the initial diagnosis; iv) had insufficient clinical follow-up data (defined as <12 months post-surgery for surviving patients); and v) the available tumor specimen was deemed inadequate for analysis due to extensive necrosis or poor preservation upon central pathological review. The patients had not received chemotherapy or radiotherapy previously. Metastasis was defined by the histological confirmation of tumors in the regional lymph nodes or distant organs at surgery. The tumor samples included 17 metastatic and 15 non-metastatic samples. The present study was approved by the Ethics Committee of the First Affiliated Hospital of Anhui Medical University (approval no. 20231337; Hefei, China) and was conducted according to the principles of the Declaration of Helsinki. Written informed consent was obtained from all patients.
Reagents
p-EGFR (phospho-Y1068; cat. no. ab40815; 1:500) and MMP7 (cat. no. ab207299; 1:1,000) were purchased from Abcam. PV-9000 histochemical reagent kit (cat. no. PV-9000) was obtained from Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. and DAB (cat. no. P0201S) staining solution was obtained from Beyotime Biotechnology.
Bioinformatics analysis
Publicly available databases and analytical tools were employed to investigate gene expression correlations and association with survival. The profile of gene upregulation in GC was obtained from the University of Alabama at Birmingham Cancer database (UALCAN; http://ualcan.path.uab.edu/cgi-bin/TCGAExHeatMap2.pl?size=25&cancer=STAD). The transcript levels of MMP7 and EGFR were compared between tumor and adjacent normal tissues using TCGA-stomach adenocarcinoma (STAD) dataset within the UALCAN platform. The specific analysis can be replicated using the following direct links: MMP7 (https://ualcan.path.uab.edu/cgi-bin/TCGAExResultNew2.pl?genenam=MMP7&ctype=STAD) and EGFR (https://ualcan.path.uab.edu/cgi-bin/TCGAExResultNew2.pl?genenam=EGFR&ctype=STAD). To ensure full reproducibility of the present analyses, a comprehensive step-by-step protocol is provided in Data S1. Kaplan-Meier plotter (https://kmplot.com/analysis/) was used to examine the associations between MMP7 or EGFR expression and overall survival. All available datasets in the platform were included without any dataset-specific restrictions. Cut-off values for gene expression were determined automatically based on percentiles. Specifically, the threshold corresponding to the 70th percentile was used for MMP7 (absolute expression value, 1,486; range, 5–37,183), and the threshold corresponding to the 75th percentile was used for EGFR (absolute expression value, 53; range, 1–880). These thresholds were applied to stratify patients into high and low expression groups for subsequent survival analysis.
IHC analysis
Paraffin-embedded tissue sections were used for immunohistochemistry. The source tissues had been previously fixed in 10% neutral buffered formalin at room temperature for 24–48 h during routine pathological processing. For IHC, 5 µm sections were mounted on positively charged slides and dried in an oven at 60°C for 1 h prior to staining. Tissue slides were deparaffinized using xylene (100%, 15 min, 25°C, twice) and rehydrated through a graded ethanol series (100% ethanol, 3 min, 25°C, twice; 95% ethanol, 3 min, 25°C; 90, 80 and 70% ethanol, 1 min each, 25°C). The tissue slices were soaked in citrate buffer (0.01 M, 100°C) for antigen retrieval. After boiling for 15 min, the samples were allowed to cool. The effects of endogenous enzymes were eliminated after 20 min of treatment with H2O2 (reagent 1, 25°C). Serum (10%) blocking was performed at 25°C for 20 min, followed by incubation with p-EGFR or MMP7 primary antibodies at 4°C for 16 h. Reaction-boosting solution (reagent 2) and secondary antibody (reagent 3) were added sequentially and incubated for 20 min at 25°C. Reagents 1, 2 and 3 were included in the PV-9000 histochemical kit. DAB was used for 5 min for color development. Nuclei were stained with hematoxylin (Beyotime Biotechnology) for 3 min at 25°C and sealed with neutral glue. Staining was independently assessed by two experienced pathologists who were blinded to the clinical data. Inter-observer variability was quantitatively evaluated using Cohen's κ coefficient. A light Panoramic MIDI scanner with Jetta JD801 (Jiangsu Jetta Technology Development Co., Ltd.) was used to capture the images. A combined score of staining intensity and distribution was used to semi-quantitatively evaluate p-EGFR and MMP7 expression (37). For comparative analysis, patients were stratified into low-(lowest 30%), moderate- and high-(highest 30%) expression cohorts. The average optical density was analyzed using ImageJ software (version 1.44p; National Institutes of Health).
Hematoxylin-eosin (H&E) staining
Paraffin-embedded tissues were cut into 5 µm sections and mounted on positively charged slides and dried in an oven at 60°C for 1 h prior to staining. Tissue slides were deparaffinized using xylene (100%, 15 min, 25°C, twice) and rehydrated through a graded ethanol series (100% ethanol, 3 min, 25°C, twice; 95% ethanol, 3 min, 25°C; 90, 80 and 70% ethanol, 1 min each, 25°C). The nuclei were stained with hematoxylin for 3 min at 25°C and the cytoplasm was stained with eosin for 1 min at 25°C (Beyotime Biotechnology). Images were captured using a panoramic scanner (Panoramic MIDI).
Statistical analysis
IHC staining for MMP7 or EGFR was performed on sequential sections from 32 tissue samples, with each staining procedure repeated in triplicate to ensure reproducibility. The average optical density values were quantified using ImageJ software. All data are presented as the mean ± SD. The primary statistical analyses, including between two groups comparisons using paired or unpaired two-tailed Student's t-tests and correlation analysis using Pearson's correlation coefficient, were performed using GraphPad Prism (version 8; Dotmatics), whereas the results for Fig. 1, Fig. 2, Fig. 3 were obtained directly from TCGA analysis portal of the UALCAN database without further modification by the authors. P<0.05 was considered to indicate a statistically significant difference.
ResultsBioinformatics analysis of MMP7 expression in GC
Gastric adenocarcinoma represents the primary form of GC, with metastasis in patients with gastric adenocarcinoma being a key determinant of their quality of life and survival. The present study analyzed the top 25 upregulated genes in gastric adenocarcinoma (Normal, n=34; Tumor, n=415) using data from TCGA database and identified the expression levels of MMP11 and MMP7 elevated (both MMP7 and MMP11 were members of the MMPs family; Fig. 1A). MMP7, specifically, is secreted by cancer cells and possesses the ability to degrade the ECM, facilitating the breach of the initial defensive barrier during cancer cell metastasis.
The transcript levels of MMP7 in gastric adenocarcinoma tissue samples (n=415) were significantly higher compared with those in normal tissue samples (n=34; P=1.620×10−12; Fig. 1B). Furthermore, the survival prognosis curve revealed that increased MMP7 expression was significantly associated with unfavorable survival outcomes in patients (Fig. 1C; P=2.000×10−2).
Next, the stage-specific analysis of MMP7 expression in gastric adenocarcinoma revealed significantly elevated levels across all tumor grades compared with that in normal tissues (Grade 1, P=3.780×10−2, n=12; Grade 2, P=5.006×10−8, n=148; Grade 3, P=1.209×10−10, n=246), demonstrating progressively increasing expression from well-differentiated to poorly-differentiated tumors, although inter-grade differences did not reach statistical significance (Fig. 2A). The present study findings revealed that MMP7 is consistently upregulated in gastric adenocarcinoma regardless of tumor grade, with a non-significant increase accompanying loss of differentiation. The pattern supports its involvement in both tumor initiation and progression. The findings suggest that MMP7 upregulation occurs early in gastric adenocarcinoma development and persists throughout tumor progression, supporting its potential role as a consistent molecular marker across different disease stages.
Furthermore, the present comprehensive analysis revealed significant MMP7 upregulation across all clinical stages of gastric adenocarcinoma compared with that in normal tissues (n=34), with stage-specific elevations observed in Stage 1 (n=18; P=1.709×10−2), Stage 2 (n=123; P=7.797×10−7), Stage 3 (n=169; P=1.297×10−7) and Stage 4 (n=41; P=8.788×10−4) tumors. Notably, while MMP7 expression progressively increased from the early to advanced stages, the inter-stage comparisons did not reach statistical significance (Fig. 2B). The significant upregulation of MMP7 across all clinical stages, compared with that in normal tissue, establishes its broad association with gastric tumorigenesis. The non-significant increasing trend with disease progression possibly reflects the function of MMP7 as a sustained driver of tumor aggressiveness. The consistent increase across all stages underscores the potential utility of MMP7 as a reliable biomarker and therapeutic target throughout the disease continuum.
After these analyses, the present study examined the disparities in MMP7 levels in the gastric adenocarcinoma tissues of male and female patients. The results indicated that compared with normal tissues (n=34), MMP7 expression was elevated in both male (n=268, P=2.148×10−11) and female (n=147, P=1.996×10−6) patients, although no notable differences were observed in the expression levels between male and female patients, as shown in Fig. 2C.
The present study compared the MMP7 expression differences in patients with gastric adenocarcinoma and lymph node metastasis. The findings highlighted that compared with normal tissues (n=34), MMP7 expression increased significantly in N0 (n=123; P=5.807×10−7), N1 (n=112; P=4.080×10−5), N2 (n=79; P=4.224×10−4) and N3 (n=82; P=3.239×10−4), with no statistically significant differences in the MMP7 expression levels between lymph node metastasis grades (N0-N3), as depicted in Fig. 2D. The observations underscored the key role of MMP7 in the metastasis of gastric adenocarcinoma and emphasized the clinical significance of selecting MMP7 as a target for further exploration and potential intervention.
Bioinformatics analysis of EGFR expression in GC
Emerging evidence indicates that MMP7 expression is modulated by upstream regulatory pathways (38,39). Notably, EGFR-mediated regulation of MMP7 has been reported not only in diabetic kidney disease (40), but also in the context of GC metastasis (41), suggesting a potentially conserved mechanism across inflammatory and malignant conditions. The present bioinformatic analysis of TCGA dataset revealed significant differential expression level of EGFR between normal gastric tissue and gastric adenocarcinoma, with notable upregulation observed in malignant tissues (P=1.608×10−2; Fig. 3A). Survival analysis demonstrated that elevated EGFR expression is significantly associated with worse clinical outcomes in patients with gastric adenocarcinoma (P=2.300×10−3; Fig. 3B), highlighting its prognostic relevance in GC progression.
Expression levels of MMP7 and p-EGFR are higher in metastatic gastric adenocarcinoma tissues
The present study utilized 32 matched pairs of gastric adenocarcinoma and adjacent normal tissue samples, including 17 metastatic and 15 non-metastatic cases. All specimens underwent comprehensive histological evaluation using H&E staining, coupled with IHC analysis of p-EGFR and MMP7 expression. Fig. 4A displays representative IHC results from 2 non-metastatic (patients 1 and 2) and 2 metastatic (patients 3 and 4) cases.
Using GraphPad Prism, the present study performed a comparative analysis of the protein expression patterns between the metastatic and non-metastatic groups. The results demonstrated a significantly elevated expression levels of both p-EGFR (P<0.0001) and MMP7 (P<0.0001) in metastatic tissues compared with that in the non-metastatic counterparts (Fig. 4B and C). The experimental findings provided notable evidence for the involvement of p-EGFR and MMP7 in gastric adenocarcinoma metastasis.
p-EGFR and MMP7 are positively correlated in gastric adenocarcinoma tissue
IHC analysis of 17 metastatic GC cases revealed differential MMP7 expression levels, ranging from low (patient 5), moderate (patient 6) to high (patient 7) staining intensity (Fig. 5A). Comparative evaluation of all 32 matched tumor-normal tissue pairs demonstrated significantly elevated expression levels of both p-EGFR (P<0.0001) and MMP7 (P<0.0001) in gastric adenocarcinoma tissues compared with that in their normal counterparts (Fig. 5B and C).
Notably, as shown in Fig. 5A, as the MMP7 expression level decreased (as seen in patient 5), p-EGFR expression decreased. By contrast, when MMP7 expression was high (as observed in patient 7), p-EGFR expression levels increased. Therefore, the present study performed a correlation analysis between p-EGFR and MMP7 expression in 32 gastric adenocarcinoma tissue samples and revealed a positive correlation between p-EGFR and MMP7 expression (r2=0.6219; P<0.0001), as shown in Fig. 5D. Furthermore, a correlational analysis of EGFR and MMP7 expression, conducted via the Gene Expression Profiling Interaction Analysis platform (https://gepia3.bioinfoliu.com/), identified a statistically significant yet weak positive relationship (r=0.27; P=4.4×10−12; Fig. S1). The correlation underscores the association between p-EGFR and MMP7 in gastric adenocarcinoma, reinforcing their potential relevance in disease progression.
Discussion
GC is one of the most lethal malignancies worldwide and ranks among the leading causes of cancer-related mortality (42). Its aggressive nature, characterized by its pronounced invasive and metastatic potential, poses notable clinical challenges. Although therapeutic advances over the past decade have improved patient outcomes, persistent issues of post-treatment recurrence and metastasis continue to compromise patient prognosis. The unresolved clinical challenges underscore the key need to identify the molecular targets involved in GC metastasis, a key research priority that could markedly enhance postoperative survival rates and quality of life in affected individuals.
MMPs are key mediators of tumor metastasis, drawing notable research interest due to their multifaceted roles in both physiological and pathological processes. They were initially characterized for their functions in embryonic development and tissue remodeling (43,44). However, they are now recognized as key contributors to cancer pathogenesis due to their ability to degrade basement membrane components and ECM proteins (45). This proteolytic activity facilitates key oncogenic processes, including tumor invasion, angiogenesis and metastatic dissemination. Previous studies have reported that the activity and expression levels of MMPs, such as MMP2, MMP3, MMP7 and MMP9, are increased in patients with GC and can reduce their survival period, promote the metastasis and recurrence of cancer and render a poor prognosis (46–48).
MMP7 exhibits unique clinical value as a tumor-derived protease, thereby being distinct from other MMP family members that are primarily secreted by stromal cells. This tumor cell-specific expression pattern makes MMP7 an ideal biomarker in monitoring cancer progression. Increasing evidence demonstrates MMP7 upregulation in multiple malignancies, including prostate and breast cancer, and its association with metastasis (49,50). MMP7 can be used as a marker to measure the prognosis of colon and esophageal cancer (51). Using bioinformatics analysis, the present study demonstrated that MMP7 is upregulated in gastric adenocarcinoma and is associated with a worse patient prognosis. Notably, the IHC results of the present study revealed that MMP7 was highly expressed in gastric adenocarcinoma tissues with lymph node metastasis, which demonstrated that MMP7 is associated with cancer progression and is a tumor marker for the invasion, metastasis and poor prognosis of GC. Therefore, it could be a potential therapeutic target for the treatment of GC.
EGFR, a key member of the human EGFR family, initiates downstream signaling cascades (for example, the PI3K/AKT, STAT and MAPK pathways) upon ligand binding and phosphorylation. These pathways notably regulate tumor cell survival, apoptosis, invasion and metastasis (52–55). Aberrant EGFR activation in GC is strongly implicated in the promotion of metastatic progression (24,56). Emerging evidence indicates that EGFR can directly modulate MMP7 transcription, as in lung cancer progression (57,58). Using TCGA database analysis, the present study identified that the upregulation of EGFR in GC is associated with a worse prognosis of patients, consistent with a previous report. In addition, IHC results suggested that p-EGFR and MMP7 were both expressed in clinical GC samples. Notably, the present study identified that p-EGFR and MMP7 are positively correlated in GC. To definitively establish causality and elucidate the mechanism associating p-EGFR with MMP7, a rigorous experimental follow-up would be required. This would include genetic manipulation (small interfering RNA/CRISPR) to confirm the functional necessity and sufficiency, pharmacological inhibition to delineate the key downstream signaling cascades (for example, MEK/ERK and PI3K/AKT) and chromatin immunoprecipitation assays to identify the transcription factors (for example, activator protien-1 and E26 transformation-specific) that directly bind to the MMP7 promoter. The current clinical evidence in the present study serves as a key foundation for such targeted mechanistic studies.
Although the present study established an association between p-EGFR and MMP7 in GC, the precise regulatory mechanisms remain to be elucidated due to platform constraints. Further investigation is warranted to elucidate a few points. First, the association between p-EGFR and MMP7, although significant in the present study, would require mechanistic validation; whether p-EGFR directly regulates MMP7 transcription and which specific signaling intermediates are involved remain to be elucidated. Second, the prognostic and therapeutic potential of this axis has not yet been fully defined. Future efforts should focus on prospectively validating the combined p-EGFR/MMP7 profile as a clinical biomarker for patient stratification and evaluation of the efficacy of targeting this pathway. Lastly, the relatively short follow-up period of the present study cohort limited a robust overall survival analysis. Therefore, expanding the present study to a larger, independent validation cohort with longer follow-up period is a priority. Furthermore, investigating the potential interplay among MMP7, p-EGFR and other upregulated proteins (for example, collagen type I α 1 chain, ubiquitin D and cystatin 1) would be a compelling avenue for future studies.
In summary, the present study systematically elucidated the synergistic mechanism of p-EGFR and MMP7 in GC using an integrated analysis of TCGA database and IHC detection of clinical samples. The key findings were as follows: i) Significant upregulation of both p-EGFR and MMP7 in GC tissues, which were significantly associated with poor patient prognosis and metastatic progression; and ii) to the best of our knowledge, IHC results demonstrated for the first time a significant positive correlation between p-EGFR and MMP7 expression in GC tissues. The findings not only provide novel experimental evidence in understanding the molecular mechanisms of GC metastasis, but, more notably, establish the p-EGFR/MMP7 signaling axis as a potential dual target for metastatic GC treatment, laying a key theoretical foundation for the development of novel targeted therapies in the future.
Supplementary Material
Supporting Data
Acknowledgements
Not applicable.
Availability of data and materials
The data generated in the present study may be requested from the corresponding author.
Authors' contributions
BD and YWa conceived the present study, participated in data analysis and drafted and wrote the manuscript. YWu, ZZ and YM collected clinical tissue samples and conducted immunohistochemical experiments. ZW, RJ and TL conceived the study, led its design and contributed to the revision of the manuscript. All authors read and approved the final version of the manuscript. BD, YWa, YWu, ZZ, YM, ZW, RJ and TL confirm the authenticity of all the raw data.
Ethics approval and consent to participate
The present study was conducted in accordance with the Declaration of Helsinki and was approved by the Institutional Review Board of First Affiliated Hospital of Anhui Medical University (Hefei, China; approval no. 20231337). Written informed consent was obtained from all participants involved in the study.
Patient consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
AbbreviationsGC
gastric cancer
IHC
immunohistochemical
EGFR
epidermal growth factor receptor
MMPs
matrix metalloproteinases
ECM
extracellular matrix
H&E
hematoxylin-eosin
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Bioinformatics analysis of MMP7 expression in gastric cancer. (A) Gene profile of upregulation in GC, showing the top 25 upregulated genes. (B) TCGA analysis: MMP7 expression in GC and normal tissue. The error bars indicate the non-outlier data range, the box shows the interquartile range and the internal line marks the median. Individual dots represent outliers. (C) Survival prognosis of patients with GC based on MMP7 expression: An analysis of TCGA data. MMP7, matrix metalloproteinase 7; GC, gastric cancer; TCGA, The Cancer Genome Atlas; STAD, stomach adenocarcinoma; HR, hazard ratio; TPM, transcripts per million.
Figure 1. Bioinformatics analysis of MMP7 expression in gastric cancer. (A) Gene profile of upregulation in GC, showing the top 25 upregulated genes. (B) TCGA analysis: MMP7 expression in GC and norma...
Bioinformatics analysis of MMP7 expression in gastric cancer. (A) Expression level of MMP7 in different stages of GC. (B) Expression level of MMP7 in different grades of GC and normal tissues. (C) Expression level of MMP7 in male and female patients with GC. (D) Expression level of MMP7 in different lymph node states of GC. For all box plots, error bars depict the variability of the data outside the interquartile range, extending to the farthest non-outlier data points. The box shows the interquartile range and the internal line marks the median. MMP7, matrix metalloproteinase 7; GC, gastric cancer; STAD, stomach adenocarcinoma.
Figure 2. Bioinformatics analysis of MMP7 expression in gastric cancer. (A) Expression level of MMP7 in different stages of GC. (B) Expression level of MMP7 in different grades of GC and normal tissue...
Bioinformatics analysis of EGFR expression in gastric cancer. Upregulation of EGFR in gastric adenocarcinoma is associated with worse patient prognosis. (A) Expression level of EGFR between normal and GC samples. The box plot shows the median, interquartile range and full data range (error bars). Individual data points are overlaid. (B) Analysis of survival prognosis of EGFR in GC. EGFR, epidermal growth factor receptor; TCGA, The Cancer Genome Atlas; STAD, stomach adenocarcinoma; HR, hazard ratio.
Figure 3. Bioinformatics analysis of EGFR expression in gastric cancer. Upregulation of EGFR in gastric adenocarcinoma is associated with worse patient prognosis. (A) Expression level of EGFR between ...
Expression levels of MMP7 and p-EGFR are higher in metastatic gastric adenocarcinoma tissues. (A) IHC and H&E analysis for the differential expression levels of p-EGFR and MMP7 in metastatic and non-metastatic GC and normal tissues. Patients 1–2 were classified as non-metastatic, while patients 3–4 were categorized as metastatic (scale bar, 200 µm). (B) Quantitative analysis of p-EGFR expression (AOD) using IHC staining between metastatic (n=17) and non-metastatic (n=15). ****P<0.0001. (C) Quantitative analysis of MMP7 expression (AOD) using IHC staining between metastatic (n=17) and non-metastatic (n=15). ****P<0.0001. Error bars represent the SD from the mean of three independent experiments. MMP7, matrix metalloproteinase 7; p-EGFR, phosphorylated-epidermal growth factor receptor; GC, gastric cancer; AOD, average optical density; IHC, immunohistochemical; H&E, hematoxylin-eosin.
Figure 4. Expression levels of MMP7 and p–EGFR are higher in metastatic gastric adenocarcinoma tissues. (A) IHC and H&E analysis for the differential expression levels of p–EGFR and MMP7 in metastatic...
p-EGFR and MMP7 are positively correlated in gastric adenocarcinoma tissue. (A) IHC analysis for p-EGFR and MMP7 expression and H&E staining in GC tissues and normal tissues (scale bar, 200 µm). (B) Quantitative analysis of p-EGFR expression (AOD) using IHC staining between GC and corresponding normal tissues. (C) Quantitative analysis of MMP7 expression (AOD) using IHC staining between GC and corresponding normal tissues. (D) Correlation between p-EGFR and MMP7 expression in GC tissues, with the orange part representing a 95% confidence interval. ****P<0.0001. Error bars represent the SD from the mean of three independent experiments. MMP7, matrix metalloproteinase 7; p-EGFR, phosphorylated-epidermal growth factor receptor; GC, gastric cancer; AOD, average optical density; IHC, immunohistochemical; HE, hematoxylin-eosin.
Figure 5. p–EGFR and MMP7 are positively correlated in gastric adenocarcinoma tissue. (A) IHC analysis for p–EGFR and MMP7 expression and H&E staining in GC tissues and normal tissues (scale bar, 200 ...