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Tumor Discovery MMP-1 as a potential biomarker for gastric cancer

2.4. Cell migration was correlated with the malignant phenotype of gastric

During the logarithmic growth phase, N87 cells were cancer cell lines (Figure 1). A search of The Cancer Genome
inoculated into a 6-well plate. When the cell growth Altlas database revealed the presence of enhanced MMP-1
rate reached 80%, Ad-MMP-1, the two interference expression or mutations in various malignancies. Based on
plasmids and the corresponding controls were transfected 170 gastric cancer-related studies involving 4067 patients,
with Lipofectamine 3000 at specific concentrations. MMP-1 polymorphisms were found to be associated
Subsequently, a 200-µL pipette tip was used to make with gastric cancer. Previous studies have linked elevated
scratches, and the medium was replaced with a new medium MMP-1 expression levels with patient prognosis across
containing 1% serum. Images were captured to document different malignancies (Figure 2A). In the present study,
the initial (0-h) scratch data. Following incubation under fluorescence quantitative polymerase chain reaction (PCR)
low-serum culture conditions for 48 h, the scratch was assays were performed on collected EGC and control
imaged to record the 48-h data. The changes in the scratch- tissue samples. MMP-1 expression was significantly higher
wound area and the differences between the groups were in EGC tissues than in their paracancerous counterparts
calculated and compared using the ImageJ software. (Figure 2B). Further analysis indicated that high MMP-1
expression has a sensitivity and specificity of 0.67 and 0.8,
2.5. Western blotting respectively, for detecting EGC, suggesting its potential as
Total protein was extracted from transfected N87 cells a biomarker for EGC diagnosis (Figure 2C).
using RIPA lysis buffer, and their concentrations were
measured using a BCA kit. Then, the cells were mixed 3.2. Effect of MMP-1 on gastric cancer cell
with SDS buffer for 5 min at 100°C to prepare the proliferation
samples. After SDS-PAGE electrophoresis, the proteins The upregulation of MMP1 in N87 cells was verified by
were transferred to polyvinylidene difluoride membranes western blotting (Figure 3A). EdU experiments were
using electrophoresis equipment. Immunoreactivity was performed on N87 cells transfected with Ad-MMP-1 and
measured after blocking with 5% skim milk. Images were the respective control groups. The ratio of EdU-positive
captured using a chemiluminescence imaging system, and cells was significantly higher in the MMP-1 overexpression
any differences observed were analyzed. group than in the control group (Figure 3B), indicating
a significant improvement in the proliferative ability of
2.6. Statistical methods
N87 cells by MMP-1 overexpression. In addition, MMP1
GraphPad 9.0 software was utilized to conduct all statistical downregulation was verified by western blot analysis
analyses of the data. The mean values of the two groups (Figure 4A). Simultaneously, N87 cells with MMP-1
were compared using an independent-sample t-test. knockdown were examined using EdU experiments,
A P < 0.05 indicated a significant difference. revealing a decreased proportion of EdU-positive cells and

3. Results a significantly diminished proliferative ability, particularly
evident in the sh2 group (P < 0.05; Figure 4B).
3.1. MMP-1 as the potential diagnostic marker for
EGC 3.3. Effect of MMP-1 on gastric cancer cell migration
After whole-transcriptome RNA sequencing and Results of the scratch experiment analysis revealed that
subsequent bioinformatics analysis, three pairs of early the scratch wound area of N87 cells with upregulated
and advanced gastric cancer cells, along with their MMP-1 expression was reduced by 62.3% compared
paracancerous tissues, were derived using clinical to the initial scratch area at 48 h. This represented a
endoscopy. Pathological examination confirmed the significant increase compared to that in the control
samples, which were then subjected to analysis. Nearly group; specifically, a 44% reduction in the scratch wound
1,831 differentially expressed coding genes were present area was observed with a P-value of 0.0047 (Figure 5A).
in advanced gastric cancer, whereas there were only 121 In contrast, the scratch wound areas in the sh1 and
differentially expressed coding genes in EGC (Figure 1A). sh2 (RNA interference) groups were reduced by 46.8%
A heat map revealed 44 differentially expressed genes and 48.6%, respectively. This represented a significant
in both early and advanced gastric cancer, of which 17 decrease compared with the percentage reduction in the
were upregulated, including MMP-1 (Figure 1B). The corresponding control group (60.6%). These differences
membrane surface protein molecule, MMP-1, was initially were statistically significant, with P-values of 0.0009 and
screened due to its higher expression levels in both early 0.004, respectively (Figure 5B), suggesting that MMP-1
and advanced gastric cancer tissues. Its expression level promoted cell migration.

Volume 3 Issue 1 (2024) 3 https://doi.org/10.36922/td.1973