B-cell lymphoma 9 (Bcl9) is the core component of Wnt/β-catenin signaling and overexpressed in nuclei of various tumors, including hepatocellular carcinoma (HCC). However, the extent of Bcl9 expression relative to HCC differentiation stage and its functional aspects are poorly understood. In this study, we examined the expression pattern of Bcl9 immunohistochemically, using two anti-Bcl9 antibodies; one was a conventional polyclonal-antibody (anti-Bcl9ABC) against amino acid no.800-900 of human-Bcl9, while the other (anti-Bcl9BIO) was against amino acid no.50-200, covering Pygopus-binding sites of Bcl9. Immunohistochemistry using anti-Bcl9BIO demonstrated distinctive staining in the cytoplasm, while the anti-Bcl9ABC signal was detected in both cytoplasm and nuclei of HCC cells, reflecting different states of Bcl9 function because Pygopus-binding to Bcl9 is essential to exert its function together with β-catenin in nucleus. Quantitative analysis revealed a significantly higher immunohistochemical-score by anti-Bcl9BIO in normal liver comparing various differentiation grades of HCC (P < 0.004), whereas no significant difference was noted with anti-Bcl9ABC. Interestingly, immunohistochemical-score of anti-Bcl9BIO in patients aged < 40 years was significantly lower than that of ≥ 40 years group (P < 0.01). The results indicated that anti-Bcl9BIO detected cytoplasmic Bcl9, which does not bind to Pygopus suggesting it could be a useful indicator for development of HCC in young Myanmar patients.

中文翻译：

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Bcl9的免疫组织化学定位使用识别不同表位的两种抗体可用于表征缅甸肝细胞癌的青少年发展。

B细胞淋巴瘤9（Bcl9）是Wnt /β-catenin信号转导的核心成分，在包括肝细胞癌（HCC）在内的各种肿瘤的核中过表达。但是，Bcl9表达相对于肝癌分化阶段的程度及其功能方面了解甚少。在这项研究中，我们使用两种抗Bcl9抗体通过免疫组织化学检查了Bcl9的表达模式；一个是针对人Bcl9氨基酸800-900的常规多克隆抗体（anti-Bcl9ABC），而另一个（anti-Bcl9BIO）针对氨基酸50-200的氨基酸，涵盖了Bcl9的Pygopu​​s结合位点。使用抗Bcl9BIO进行的免疫组织化学显示细胞质中有独特的染色，而在肝癌细胞的细胞质和细胞核中均检测到抗Bcl9ABC信号，反映Bcl9功能的不同状态，因为Pygopu​​s结合Bcl9对于在细胞核中与β-catenin一起发挥其功能至关重要。定量分析显示，与各种分化程度的HCC相比，正常肝中抗Bcl9BIO的免疫组织化学评分显着更高（P <0.004），而抗Bcl9ABC则没有显着差异。有趣的是，年龄小于40岁的患者的抗Bcl9BIO免疫组化评分显着低于≥40岁的患者（P <0.01）。结果表明，抗Bcl9BIO检测到胞质Bcl9，它不与扁桃体结合，表明它可能是缅甸年轻患者肝癌发生的有用指标。定量分析显示，与各种分化程度的HCC相比，正常肝中抗Bcl9BIO的免疫组织化学评分显着更高（P <0.004），而抗Bcl9ABC则没有显着差异。有趣的是，年龄小于40岁的患者的抗Bcl9BIO免疫组化评分显着低于≥40岁的患者（P <0.01）。结果表明，抗Bcl9BIO检测到胞质Bcl9，它不与扁桃体结合，表明它可能是缅甸年轻患者肝癌发生的有用指标。定量分析显示，与各种分化程度的HCC相比，正常肝中抗Bcl9BIO的免疫组织化学评分显着更高（P <0.004），而抗Bcl9ABC则没有显着差异。有趣的是，年龄小于40岁的患者的抗Bcl9BIO免疫组化评分显着低于≥40岁的患者（P <0.01）。结果表明，抗Bcl9BIO检测到胞质Bcl9，它不与扁桃体结合，表明它可能是缅甸年轻患者肝癌发生的有用指标。40岁显着低于≥40岁组（P <0.01）。结果表明，抗Bcl9BIO检测到胞质Bcl9，它不与扁桃体结合，表明它可能是缅甸年轻患者肝癌发生的有用指标。40岁显着低于≥40岁组（P <0.01）。结果表明，抗Bcl9BIO检测到胞质Bcl9，它不与扁桃体结合，表明它可能是缅甸年轻患者肝癌发生的有用指标。