## 3 Results
### Overall Survival  
In assessing the impact of various clinicopathological factors on overall survival in CRC, our analysis revealed no significant differences based on age, gender, tumor localization, or macroscopic aspect of the tumors (Figure 3). These findings suggest that, within the cohort studied, these specific demographic and tumor characteristics do not independently influence the survival outcomes of patients. Despite the variability inherent in patient demographics and tumor presentations, these factors did not correlate with significant variations in survival rates, indicating that other molecular or genetic markers may play more critical roles in determining patient prognosis in colorectal cancer.  
The analysis indicated that neither lymph node ratio nor tumor budding significantly impacted survival outcomes. This suggests that these parameters, while indicative of tumor behavior and potential spread, do not alone dictate survival probabilities within the observed cohort. Conversely, significant differences in survival were observed when considering the microscopic aspect and pT stage of the tumors. Patients with tumors classified as G1 and G2 stages experienced better survival outcomes compared to those with G3-stage tumors, highlighting the importance of tumor differentiation as a prognostic indicator. Additionally, cases with a pT stage of less than 2 demonstrated improved survival compared to those with more advanced pT stages (Figure 4).  
Figure 3: Kaplan–Meier survival curves demonstrating the lack of significant impact of age (**A**), gender (**B**), tumor localization (**D**), and macroscopic aspect (**C**) on overall survival in CRC patients.  
In our comprehensive analysis of immunohistochemical markers and their impact on overall survival, distinct patterns emerged regarding the prognostic significance of ARSB, Maspin, and TP53 protein expressions. Positivity for ARSB, as determined by immunohistochemistry, was associated with better survival outcomes, suggesting its potential role as a protective factor in CRC progression. Similarly, Maspin positivity was linked to improved survival, indicating that its expression might confer a survival advantage, possibly through its effects on tumor suppression and the inhibition of metastasis. However, the immunohistochemical expression of TP53 showed no significant difference in survival (Figure 5).  
In the exploration of gene expression profiles and their correlation with survival outcomes, our study unveiled noteworthy trends regarding the collective expression of ARSB, TP53, and Maspin. Analysis revealed that patients exhibiting a relative quantification greater than 1 for both ARSB and TP53 genes experienced significantly better survival, suggesting a synergistic effect of these genes in enhancing patient prognosis. Similarly, a combination of TP53 and Maspin genes with RQ > 1 was associated with improved prognostic outcomes, indicating the potential of these markers in predicting favorable survival. Most compellingly, our analysis demonstrated that patients with overexpression (RQ > 1) of all three genes--ARSB, TP53, and Maspin--showed the best survival outcomes,  
Figure 4: Kaplan–Meier curves highlighting the prognostic significance of microscopic aspect (**A**) and pT stage (**C**), with lymph node ratio (**B**) and budding (**D**) showing no significant impact.  
Figure 5: Immunohistochemistryal analysis of ARSB (**A**) and Maspin (**B**) impact on survival: significant prognostic value contrasted with TP53’s (**C**) non-significant influence.  
underscoring the powerful prognostic value of these combined gene expressions in CRC. Conversely, a scenario where all three genes had RQ \(<\) 1 was linked to poorer survival, highlighting the critical impact of these genes' underexpression on patient prognosis (Figure 6).