Immunohistochemical Mapping of Bcl9 Using Two Antibodies that Recognize Different Epitopes Is Useful to Characterize Juvenile Development of Hepatocellular Carcinoma in Myanmar
Immunohistochemical Mapping of Bcl9 Using Two Antibodies that Recognize Different Epitopes Is Useful to Characterize Juvenile Development of Hepatocellular Carcinoma in Myanmar
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DOI：
10.1267/ahc.18045
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发表时间：
2019-02
影响因子：
2.4
通讯作者：
Myat Thu Soe;Y. Shibata;Myo Win Htun;K. Abe;Kyaw Soe;Nay Win Than;Thann Lwin;Myat Phone Kyaw;T. Koji
中科院分区：
文献类型：
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作者：
Myat Thu Soe;Y. Shibata;Myo Win Htun;K. Abe;Kyaw Soe;Nay Win Than;Thann Lwin;Myat Phone Kyaw;T. Koji
b细胞淋巴瘤9 （Bcl9）是Wnt/β-catenin信号传导的核心成分，在包括肝细胞癌（HCC）在内的多种肿瘤细胞核中过表达。然而，Bcl9在HCC分化阶段的表达程度及其功能方面尚不清楚。在本研究中，我们使用两种抗Bcl9抗体免疫组织化学方法检测了Bcl9的表达模式；一种是常规的多克隆抗体（抗bcl9abc）。另一种（抗bcl9bio）是针对第1号氨基酸的。50-200，覆盖Bcl9的pygopus结合位点。抗bcl9bio免疫组化在细胞质中显示出不同的染色，而抗bcl9abc信号在HCC细胞的细胞质和细胞核中均检测到，这反映了Bcl9功能的不同状态，因为pygopus与Bcl9结合是与细胞核中β-catenin一起发挥其功能所必需的。定量分析显示，与不同分化级别的HCC相比，正常肝脏中抗bcl9bio免疫组化评分显著高于正常肝脏（P < 0.004），而抗bcl9abc免疫组化评分无显著差异。有趣的是，< 40岁组抗bcl9bio免疫组化评分明显低于≥40岁组（P < 0.01）。结果表明，抗bcl9bio检测到细胞质Bcl9，而Bcl9不与Pygopus结合，这表明它可能是缅甸年轻患者HCC发展的有用指标。
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.