## 4 Discussion  
This study explores the intricate relationship between clinicopathological features, immunohistochemical expressions, gene expression profiles, and their collective impact on patient survival. This section draws upon the existing literature to contextualize our findings, shedding light on the complex interplay of molecular markers in CRC progression and prognosis.  
Our results indicated no significant correlation between TP53 gene expression and a range of clinicopathological features, including age, gender, tumor localization, and macroscopic and microscopic aspects. This lack of association stands in contrast to the significant correlation observed between TP53 gene expression and its immunohistochemical expression, suggesting that while TP53 protein levels might reflect tumor behavior, its gene expression alone does not directly correlate with the traditional clinicopathological indicators of CRC. This finding aligns with research suggesting that TP53 mutations, rather than expression levels, might play a more pivotal role in CRC pathogenesis [9; 10; 11; 12; 13].  
Significant differences in ARSB gene expression were found related to age, macroscopic aspect, and microscopic aspect, underscoring the gene's potential role in the biological aggressiveness of CRC. Specifically, higher ARSB expression in infiltrative, ulcero-infiltrating tumors, and its decreased expression in G3-stage cases, suggests ARSB's involvement in tumor differentiation and invasion, akin to findings from previous studies [2; 14; 15; 16]. Moreover, the significant correlation of ARSB expression with tumor budding, and its immunohistochemical expression alongside Maspin and TP53, further emphasizes its intricate role in CRC's molecular landscape, possibly through modulation of the tumor microenvironment [1; 2].  
Maspin gene expression presented noteworthy associations with age, tumor budding, and immunohistochemical expressions of Maspin, ARSB, and TP53, particularly highlighting its decreased expression in aggressive tumor behaviors (e.g., G3 budding). This pattern indicates Maspin's potential protective role against CRC progression, a notion supported by the literature that links Maspin with tumor suppression and apoptosis [17; 18]. The observed discrepancies in Maspin expression relative to ARSB and TP53 immunohistochemistry point to complex regulatory mechanisms influencing CRC dynamics, warranting further investigation into their potential therapeutic implications [1; 8; 19].  
Figure 6: Enhanced survival in linked to dual and triple gene expression elevations: ARSB + TP53 (**A**) and TP53 + Maspin (**B**) combinations, and high ARSB, TP53, and Maspin (**C**) expression synergy.  
The survival analysis illuminated the prognostic significance of microscopic aspect and pT stage, with poorer outcomes associated with G3 classification and pT > 3 stage, respectively. These findings are consistent with the established understanding that a higher tumor grade and advanced invasion depth are indicative of aggressive disease and worse prognosis [20]. Interestingly, while ARSB and Maspin were negativity correlated with reduced survival, TP53 immunohistochemistry alone did not exhibit a significant survival impact. This divergence underscores the complexity of TP53 role in CRC survival, possibly related to the diverse functional consequences of TP53 mutations [4]. Different expression profiles of TP53 gene highlight the possibility of mutation in the gene itself.  
Notably, dual high gene expression (ARSB + TP53 and TP53 + Maspin) and triple high gene expression (ARSB + TP53 + Maspin) were associated with significantly better survival. These results suggest a synergistic effect of these genes in modulating CRC outcomes, a novel insight that could pave the way for multifaceted molecular targeting in CRC therapy [1, 2, 11, 15, 21]. The superior survival linked to combined gene overexpression highlights the importance of a comprehensive molecular profiling approach in CRC prognostication and treatment planning.