Kepuladze Shota, Burkadze George, Kokhreidze Irakli
Pathology and Oncology, Tbilisi State Medical University, Tbilisi, GEO.
Molecular Pathology, Tbilisi State Medical University, Tbilisi, GEO.
Cureus. 2024 Sep 6;16(9):e68761. doi: 10.7759/cureus.68761. eCollection 2024 Sep.
Background Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer characterized by the lack of expression of estrogen and progesterone receptors and the absence of HER2 protein overexpression or gene amplification. How TNBC becomes so aggressive at the molecular level is not yet fully understood. The epithelial-mesenchymal transition (EMT) has been increasingly recognized as playing a pivotal role in cancer progression and metastasis. This study aimed to elucidate the connection between TNBC progression with EMT-related markers, including vimentin, beta-catenin, and E-cadherin. Methodology Rigorous immunohistochemical analysis was employed to assess the expression of vimentin, beta-catenin, and E-cadherin in primary tumors, tumor buds, and lymph node metastases (LNMs) from 137 cases with an invasive ductal carcinoma triple-negative phenotype diagnosed between 2018 and 2024. The EMT index, which was especially important in our work, is the sum of vimentin and beta-catenin expression divided by that of E-cadherin. Estimated Pearson correlation, multiple linear regression, and Kruskal-Wallis tests were used to determine the relationships of the EMT index with tumor buds and tumor-infiltrating lymphocytes (TILs). Results Vimentin highly correlated within separate regions of interest with Pearson correlation ranging from 0.90 to 0.92 (p < 0.001). Strong negative correlations between E-cadherin and vimentin (r = -0.81 to - 0.89, p < 0.001) showed its role in preserving the epithelial phenotype. The presence of tumor buds, aggregates, or clusters of cancer cells shed from the primary tumor mass invading the connective tissue showed very strong associations with the EMT index (r = 0.91, p < 0.001). Its presence is suggestive of aggressive disease and may identify a high-risk subpopulation that may benefit from more active surveillance or adjuvant treatment. Similarly, TILs correlated inversely with the EMT index (r = -0.90, p < 0.001). The most significant predictor of the EMT index, i.e., vimentin, had a model R-squared value of 1.000 in the regression analysis. Conclusions This study brings to light the importance of EMT-related markers in TNBC progression, with special emphasis on tumor buds as possible prognostic indicators for aggressive disease. The negative correlation of TILs with the EMT index indicates that an effective immune response could antagonize EMT-mediated tumor progression. These results suggest that EMT-based treatments in TNBC should be designed from a multimarker perspective by including interactions among several markers to optimize predictions and therapeutics. The results hold the potential to set future research directions and actionable outcomes that could influence clinical utility in the battle against TNBC.
背景 三阴性乳腺癌（TNBC）是一种侵袭性很强的乳腺癌亚型，其特征是雌激素和孕激素受体缺乏表达，且不存在HER2蛋白过表达或基因扩增。TNBC在分子水平上如何变得如此具有侵袭性尚未完全明确。上皮-间质转化（EMT）在癌症进展和转移中所起的关键作用已得到越来越多的认可。本研究旨在阐明TNBC进展与EMT相关标志物（包括波形蛋白、β-连环蛋白和E-钙黏蛋白）之间的联系。
方法 采用严格的免疫组织化学分析来评估波形蛋白、β-连环蛋白和E-钙黏蛋白在2018年至2024年间诊断为三阴性表型的137例浸润性导管癌患者的原发性肿瘤、肿瘤芽和淋巴结转移灶（LNMs）中的表达。EMT指数在我们的研究中尤为重要，它是波形蛋白和β-连环蛋白表达之和除以E-钙黏蛋白的表达。使用估计的Pearson相关性、多元线性回归和Kruskal-Wallis检验来确定EMT指数与肿瘤芽和肿瘤浸润淋巴细胞（TILs）之间的关系。
结果 波形蛋白在不同感兴趣区域内高度相关，Pearson相关性范围为0.90至0.92（p < 0.001）。E-钙黏蛋白与波形蛋白之间存在强负相关性（r = -0.81至-0.89，p < 0.001），表明其在维持上皮表型中的作用。原发性肿瘤块中脱落并侵入结缔组织的癌细胞芽、聚集体或簇的存在与EMT指数显示出非常强的相关性（r = 0.91，p < 0.001）。其存在提示疾病具有侵袭性，可能识别出一个可能从更积极监测或辅助治疗中获益的高危亚群。同样，TILs与EMT指数呈负相关（r = -0.90，p < 0.001）。在回归分析中，EMT指数的最重要预测因子即波形蛋白的模型决定系数R²值为1.000。
结论 本研究揭示了EMT相关标志物在TNBC进展中的重要性，特别强调肿瘤芽作为侵袭性疾病可能的预后指标。TILs与EMT指数的负相关表明有效的免疫反应可以拮抗EMT介导的肿瘤进展。这些结果表明，TNBC基于EMT的治疗应从多标志物角度设计，包括几个标志物之间的相互作用，以优化预测和治疗。这些结果有可能为未来的研究方向和可采取行动的结果设定方向，从而可能影响对抗TNBC的临床效用。
