## 4 Discussion  
In our morphological study, we sought to substantiate or refute the hypotheses we had posited by assessing the tissue levels of four heat shock proteins, namely, Hsp10, Hsp27, Hsp60, and Hsp90, as well as scrutinizing the immunopositivity localization for each protein across various stages of differentiation within the carcinogenic vocal cord mucosa, juxtaposed with healthy tissue samples. Cells heavily rely on heat shock proteins for critical tasks essential to maintaining cellular homeostasis. Consequently, these proteins are ubiquitously present and fulfill indispensable roles, encompassing the facilitation of proper protein folding and the translocation, degradation, and elimination of aberrant protein aggregates [30]. These multifaceted functions render them chaperonins. Notably, their involvement transcends these primary duties and extends into other domains, including DNA replication [31], gene expression regulation [30; 31], the modulation of apoptotic and autophagic processes, participation in cellular senescence [32], and the induction of an inflammatory state. Remarkably, our study unveiled an unexpected association between these proteins and the intricate processes underpinning vocal cord carcinogenesis [33]. The findings of this analyzed protein pool align with existing scientific evidence that supports HSPs playing a role in the oncogenic process and its progression [34]. In several recent studies, the implication of heat shock proteins in various mechanisms underlying the onset of different types of tumors has been hypothesized and, at times, confirmed [35; 36; 37]. Gastric, hepatic, breast, ovarian, lung, pancreatic, colorectal, squamous cell of tongue, glioma, and osteosarcoma cancers are among the many tumor variants in which the involvement of heat shock proteins has been highlighted. This implication of heat shock proteins has also been found in laryngeal squamous cell carcinoma [38; 39; 40]. Heat shock proteins seem to be involved in the carcinogenesis process either directly or indirectly. Due to the hostile tumor microenvironment, cells overexpress hsp genes in response to altered oxygen levels, nutrient and metabolite deficiencies, and significant pH shifts [31]. One of the indirect mechanisms that are commonly recognized involves cancer cells releasing extracellular vesicles that contain Hsp27 and Hsp90 into the tumor environment. These heat shock proteins interact with plasma receptors on cancer cells, triggering internal signaling pathways that promote the epithelial-mesenchymal transition (EMT) process. EMT is a significant characteristic of cancer [41]. In addition, HSPs  
Figure 4: (**A**) Hsp90 immunohistochemistry. Representative images of immunohistochemical results for Healthy (**a,b**), Dysplasia (**c,d**), and SCC (squamous cell carcinoma; (**e,f**)). (**a,c,e**): magnification 200\(\times\), scale bar 50 \(\upmu\)m; (**b,d,f**): magnification 400\(\times\); scale bar 20 \(\upmu\)m. (**B**) Statistical analysis of Hsp90 results. The histogram revealed a significant difference between the Healthy group and the Dysplasia and SCC groups (#_p_\(\leq\) 0.01), as well as between the Dysplasia group and the SCC group (*_p_\(\leq\) 0.01). play a pivotal role in the regulation of extracellular matrix (ECM) remodeling and the modulation of its stiffness through their interactions with and control over a multitude of extracellular client proteins. Notably, Hsp90a, and in certain instances, Hsp60, have been identified as indispensable factors in driving the invasiveness and fostering the development of metastases in conditions such as fibrosarcoma, esophageal squamous cell carcinoma, and breast cancer cells [32]. As mentioned before, molecular chaperones can also act on a tumor level by being released into the extracellular tumor environment. This is essential to play their pro-inflammatory role, which contributes to the modulation of the tumor microenvironment. Extracellular HSPs can act as danger signals, triggering immune responses and activating various inflammatory pathways.