Division of Histology and Embryology, Department of Human Morphology and Embryology, Wroclaw Medical University, 50-368 Wroclaw, Poland.
Laboratory of Cancer Genetics, Department of Pathology, Polish Mother's Memorial Hospital Research Institute, Rzgowska 281/289, 93-338 Lodz, Poland.
Int J Mol Sci. 2023 Feb 2;24(3):2866. doi: 10.3390/ijms24032866.
The aim of the study was to evaluate the localization and intensity of RNA-binding motif single-stranded-interacting protein 3 (RBMS3) expression in clinical material using immunohistochemical (IHC) reactions in cases of ductal breast cancer (in vivo), and to determine the level of RBMS3 expression at both the protein and mRNA levels in breast cancer cell lines (in vitro). Moreover, the data obtained in the in vivo and in vitro studies were correlated with the clinicopathological profiles of the patients. Material for the IHC studies comprised 490 invasive ductal carcinoma (IDC) cases and 26 mastopathy tissues. Western blot and RT-qPCR were performed on four breast cancer cell lines (MCF-7, BT-474, SK-BR-3 and MDA-MB-231) and the HME1-hTERT (Me16C) normal immortalized breast epithelial cell line (control). The Kaplan-Meier plotter tool was employed to analyze the predictive value of overall survival of expression at the mRNA level. Cytoplasmatic RBMS3 IHC expression was observed in breast cancer cells and stromal cells. The statistical analysis revealed a significantly decreased RBMS3 expression in the cancer specimens when compared with the mastopathy tissues ( < 0.001). An increased expression of RBMS3 was corelated with HER2(+) cancer specimens ( < 0.05) and ER(-) cancer specimens ( < 0.05). In addition, a statistically significant higher expression of RBMS3 was observed in cancer stromal cells in comparison to the control and cancer cells ( < 0.0001). The statistical analysis demonstrated a significantly higher expression of mRNA in the SK-BR-3 cell line compared with all other cell lines ( < 0.05). A positive correlation was revealed between the expression of RBMS3, at both the mRNA and protein levels, and longer overall survival. The differences in the expression of RBMS3 in cancer cells (both in vivo and in vitro) and the stroma of breast cancer with regard to the molecular status of the tumor may indicate that RBMS3 could be a potential novel target for the development of personalized methods of treatment. RBMS3 can be an indicator of longer overall survival for potential use in breast cancer diagnostic process.
本研究旨在通过免疫组织化学（IHC）反应评估 RNA 结合基序单链相互作用蛋白 3（RBMS3）在临床标本中的定位和表达强度，同时在体外乳腺癌细胞系中确定 RBMS3 在蛋白和 mRNA 水平上的表达水平。此外，将体内和体外研究中获得的数据与患者的临床病理特征相关联。免疫组化研究的材料包括 490 例浸润性导管癌（IDC）病例和 26 例乳腺病组织。在 4 种乳腺癌细胞系（MCF-7、BT-474、SK-BR-3 和 MDA-MB-231）和 HME1-hTERT（Me16C）正常永生化乳腺上皮细胞系（对照）中进行了 Western blot 和 RT-qPCR。采用 Kaplan-Meier 绘图仪工具分析 表达水平对总生存期的预测价值。在乳腺癌细胞和基质细胞中观察到细胞质 RBMS3 IHC 表达。统计分析显示，与乳腺病组织相比，癌症标本中 RBMS3 表达显著降低（<0.001）。RBMS3 表达增加与 HER2（+）癌症标本（<0.05）和 ER（-）癌症标本（<0.05）相关。此外，与对照和癌细胞相比，癌症基质细胞中 RBMS3 的表达显著升高（<0.0001）。统计分析显示，与其他细胞系相比，SK-BR-3 细胞系中 mRNA 的表达显著升高（<0.05）。在蛋白质和 mRNA 水平上，RBMS3 表达之间存在正相关，与总生存期延长相关。在肿瘤分子状态方面，癌细胞（体内和体外）和乳腺癌基质中 RBMS3 表达的差异可能表明 RBMS3 可能成为开发个性化治疗方法的潜在新靶点。RBMS3 可以作为乳腺癌诊断过程中总生存期延长的指标。
