Colorectal cancer (CRC) is the third most commonly diagnosed cancer in males and the second in females, whose survival ratio and indicating biomarkers are limited. The rapid development of multiple immunofluorescences gives rise to widespread applications of this new advanced technology called multiplex immunohistochemistry (mIHC), which makes it possible to detect several fluorescent proteins on the same tumor tissue microarray (TMA) within the same time and spatial organization. By taking advantage of this mIHC technology, we detected three tumor-associated antigens (TAA) including the human epidermal growth factor receptor 2 (HER2), the cluster of differentiation 133 (CD133), the programmed death ligand-1 (PD-L1), and one immune-associated macrophage marker, the cluster of differentiation 68 (CD68) in cancer tissues versus para-carcinomatous normal tissues derived from a cohort of 84 CRC patients. All four markers were upregulated in cancer tissue compared with normal tissues. And the expressions of CD133, HER2, PD-L1, and CD68 were correlated with pathological grade, T stage, tumor size, metastasis, respectively. Accordingly, CD133 and PD-L1 could be applied as potential diagnostic biomarkers for CRC at an early stage, while the enrichment of HER2 might act as an advanced indicator in aggressive cancer status of CRC; whereas, CD68 could be potentially considered as an advanced diagnostic indicator in CRC patients, as well as a metastatic promoter in CRC-related TME. The differential expression of these four proteins, as well as their clinicopathological correlation, indicates that these four proteins could be utilized as specific diagnostic and prognostic biomarkers in CRC patients.

中文翻译：

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多重免疫组织化学表明结直肠癌中的生物标志物。

结直肠癌 (CRC) 是男性第三大最常见的癌症，女性第二大，其存活率和指示生物标志物有限。多重免疫荧光的快速发展促进了这种称为多重免疫组织化学 (mIHC) 的新型先进技术的广泛应用，该技术使得在同一时间和空间组织内检测同一肿瘤组织微阵列 (TMA) 上的多种荧光蛋白成为可能。通过利用这种 mIHC 技术，我们检测了三种肿瘤相关抗原 (TAA)，包括人表皮生长因子受体 2 (HER2)、分化簇 133 (CD133)、程序性死亡配体-1 (PD-L1) ，以及一种免疫相关巨噬细胞标记物，癌组织中的分化簇 68 (CD68) 与源自 84 名 CRC 患者队列的癌旁正常组织。与正常组织相比，所有四种标志物在癌组织中均上调。CD133、HER2、PD-L1和CD68的表达分别与病理分级、T分期、肿瘤大小、转移有关。因此，CD133和PD-L1可作为CRC早期潜在的诊断生物标志物，而HER2的富集可作为CRC侵袭性癌症状态的高级指标；而 CD68 可能被视为 CRC 患者的高级诊断指标，以及 CRC 相关 TME 的转移启动子。这四种蛋白的差异表达，以及它们的临床病理相关性，