Downregulation of ANP32B exerts anti-apoptotic effects in hepatocellular carcinoma.
Downregulation of ANP32B exerts anti-apoptotic effects in hepatocellular carcinoma.
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DOI：
10.1371/journal.pone.0177343
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Hiasa Y
中科院分区：
文献类型：
--
作者：
Ohno Y;Koizumi M;Nakayama H;Watanabe T;Hirooka M;Tokumoto Y;Kuroda T;Abe M;Fukuda S;Higashiyama S;Kumagi T;Hiasa Y
酸性（富含亮氨酸）核磷蛋白32家族成员B（ANP 32 B）是酸性核磷蛋白32（ANP 32）家族的一个高度保守的成员，对正常组织的发育至关重要。然而，它在肝细胞癌（HCC）的发展中的作用是有争议的。在这项研究中，我们阐明了ANP 32 B在肝癌细胞系和组织中的作用。使用siRNA和ANP 32 B表达质粒和慢病毒调节HCC细胞系中的ANP 32 B表达。采用实时荧光定量RT-PCR和Western blotting方法检测细胞凋亡相关蛋白的表达水平。采用实时荧光定量RT-PCR和免疫组化方法检测ANP 32 B在HCC组织中的表达。通过siRNA敲低ANP 32 B改变了HCC细胞系中凋亡相关蛋白的表达，并降低了与促凋亡剂星形孢菌素培养的HCC细胞中caspase 3和caspase 9的裂解形式的表达，但不降低caspase 8的表达。磷酸化Bad上调，而巴克下调。此外，结合并抑制Bcl-2家族的抗凋亡蛋白的ABT-737使HCC细胞对ANP 32 B沉默诱导的凋亡具有抗性。相反，ANP 32 B过表达降低Bad磷酸化并上调巴克，但不诱导凋亡，因为Bax表达下调。在HCC患者的组织中，ANP 32 B mRNA表达的低肿瘤/非肿瘤比率与晚期UICC分期相关（p = 0.032）。肝癌组织中TUNEL阳性细胞与ANP 32 B表达平行。ANP 32 B调节Bad磷酸化以及巴克和Bax表达，从而调节肝癌细胞凋亡。这些发现表明ANP 32 B作为HCC治疗靶点的潜在价值。
The acidic (leucine-rich) nuclear phosphoprotein 32 family member B (ANP32B), a highly conserved member of the acidic nuclear phosphoprotein 32 (ANP32) family, is critical for the development of normal tissue. However, its role in the development of hepatocellular carcinoma (HCC) is controversial. In this study, we elucidated the role of ANP32B in HCC cell lines and tissues. ANP32B expression in HCC cell lines was modulated using siRNA and ANP32B expression plasmids and lentiviruses. The levels of apoptosis-related proteins were analyzed by real-time RT-PCR and Western blotting. The expression of ANP32B in tissues from patients with HCC was investigated using real-time RT-PCR and immunohistochemistry. ANP32B knockdown by siRNA altered the expression of apoptosis-related proteins in HCC cell lines and reduced the expression of cleaved forms of caspase 3 and caspase 9, but not that of caspase 8, in HCC cells cultured with the pro-apoptotic agent staurosporine. Phosphorylated Bad was upregulated, whereas Bak was downregulated. Moreover, ABT-737, which binds to and inhibits anti-apoptotic proteins of the Bcl-2 family, rendered HCC cells resistant to apoptosis induced by ANP32B silencing. Conversely, ANP32B overexpression decreased Bad phosphorylation and upregulated Bak, but did not induce apoptosis because Bax expression was downregulated. In tissues from patients with HCC, a low tumor/non-tumor ratio of ANP32B mRNA expression was related to advanced UICC stage (p = 0.032). TUNEL-positive cells were observed in parallel with ANP32B expression in HCC tissues. ANP32B modulates Bad phosphorylation as well as Bak and Bax expression, resulting in regulation of apoptosis in HCC. These findings indicate the potential value of ANP32B as a therapeutic target for HCC.