Dsg2 expression in ductal carcinoma tissues
The results showed that the mRNA expression level of Dsg2 was significantly decreased in cancer tissues compared to the matched noncancerous tissues (p < 0.01, Figure 1A). The status of differentially expressed Dsg2 gene in the breast cancer tissues was calculated as the ratio of Dsg2 mRNA expression in tumor tissue to the matched normal tissue (T/N ratio).
Correlation between Dsg2 Expression and Clinicopathological Variables
To better understand the clinical significance of Dsg2 expression in breast cancer, we analyzed the correlation between Dsg2 expression and clinicopathological variables, including involvement/non-involvement lymph node, age, histological grade, vascular non-vascular involvement and tumor size (all p > 0.05, respectively Figure 1B, C, D, E, F). However, Dsg2 expression was not associated with the clinicopathological variables (Table I). ROC curve analysis was performed to evaluate the biomarker potential of desmoglein 2. The results showed that the area under curve (AUC) was equal to 0.98. This rate is at a very acceptable level, indicating the expression of this biomarker gene is suitable for diagnosing breast cancer in tissue samples. The sensitivity was 98, specificity 70, and the cutoff value 0.47 (Figure 1G).
Increased number of inflammatory nuclei and cells
In this regard, after general H&E staining, the difference between healthy tissues and tumors was observed with a light microscope. An increase in the number of nuclei was seen in tumor tissue compared to adjacent healthy tissue (Fig 2A & B).
In many patients, the ductal incidence was observed in cancer specimens, while in healthy specimens, none was observed (Fig 2D). The observation of ductal formation is consistent with lymph node involvement and metastasis in these patients.
Inflammatory cells were observed in the tumor tissue. However, no inflammatory cells were found in healthy tissue (Fig 2C). The presence of these cells indicates that the tissue is cancerous.
Comparison of breast cancer proteome with adjacent healthy tissue using SDS-PAGE technique
The study of SDS-PAGE preparations of proteins showed a difference in protein expression in the tumoral and normal tissues. The results showed differences in protein expression between these two groups. In the band range of 120 kDa, due to the molecular weight of desmoglein 2, the band difference was well observed (Figure 3).
Ultrastructure disruption and desmosome junctions
In the study of normal tissue, cell cohesion, an appropriate number of nuclei, heterochromatin state, appropriate collagen fibers, interconnected cell membranes, and milk proteins are well observed, which is consistent with the definition of a normal cell (Fig 4 A&B). In contrast, in cancer cells, an increase in the number of nuclei is observed in the euchromatin state, cell membrane rupture, cell vacuolation, and a decrease in milk proteins, which is consistent with the definition of a cancer cell (Fig 4 C&D).
Desmosome cohesion was also observed in normal cells, but desmosome disintegration was observed in tumor tissue. The results are consistent with reduced desmoglein 2 gene expression and show the importance of cellular connections in malignancy (Fig 4 E&F). To better show the change in desmosome connections in healthy and tumor cells, we focused on the image obtained. )Respectively Fig E-1 & F-1).
Nucleus in heterochromatin state 12.6μ m. Tumor cell: C. Cell membrane rupture, increase in the number of nuclei relative to healthy tissue, cell vacuolation 12.6μ m. D. The nucleus is in the euchromatin state 12.6μ m. E. Desmosomes in a healthy cell (E1- Focused image). F. Desmosomes in the cancer cell (F1- Focused image).