Gong Li, Wei Long-Xiao, Ren Pin, Zhang Wen-Dong, Liu Xiao-Yan, Han Xiu-Juan, Yao Li, Zhu Shao-Jun, Lan Miao, Li Yan-Hong, Zhang Wei
The Helmholtz Sino-German Laboratory for Cancer Research, Department of Pathology, Tangdu Hospital, The Fourth Military Medical University, Xi'an, P.R China.
Department of Nuclear Medicine, Tangdu Hospital, The Fourth Military Medical University, Xi'an, P.R China.
PLoS One. 2014 Jan 31;9(1):e87120. doi: 10.1371/journal.pone.0087120. eCollection 2014.
Glypican-3 (GPC3) has been reported to be a novel serum and histochemical marker for HCC. The positivity or negativity for GPC3 in hepatic precancerous lesions, such as dysplastic nodules (DN), has also been described. Moreover, our previous studies have demonstrated that some DN in liver cirrhosis represent monoclonal hyperplasia, and confirmed their neoplastic nature. However, additional studies must be performed to investigate further the relationship between DN with GPC3 positivity and HCC. Thus, we first investigated the expression of GPC3 in 136 HCC and 103 small DN (less than 1 cm in diameter) by immunohistochemical staining and determined the clonality of 81 DN from female patients using X-chromosome inactivation mosaicism and polymorphism of androgen receptor (AR) gene. Then we examined these samples for chromosomal loss of heterozygosity (LOH) at 11 microsatellite polymorphism sites. The results demonstrated that GPC3 immunoreactivity was detected in 103 of 136 HCC (75.7%) and 19 of 103 DN (18.4%), and the positive ratio correlated with HBsAg positivity. Clonality assays showed that 15 GPC3-positive DN from female patients, including 12 high-grade DN (HGDN), and 28 (42.4%) of 66 GPC3-negative DN, were monoclonal. In addition, among 19 GPC3-positive DN, chromosomal LOH was found at loci D6S1008 (100%, 19/19), D8S262 (52.6%, 10/19) and D11S1301 (57.9%, 11/19). However, the LOH frequency in GPC3-negative DN was 5.95% (5/84), 23.8% (20/84), and 4.76% (4/84) in three loci, respectively. Thus, we concluded that GPC3-positive DN, especially GPC3-positive HGDN, was really a late premalignant lesion of HCC.
据报道，磷脂酰肌醇蛋白聚糖-3（GPC3）是一种新型的肝癌血清和组织化学标志物。也有关于肝癌前病变（如发育异常结节，DN）中GPC3阳性或阴性的描述。此外，我们之前的研究表明，肝硬化中的一些DN代表单克隆增生，并证实了它们的肿瘤性质。然而，必须进行更多研究以进一步探究GPC3阳性的DN与肝癌之间的关系。因此，我们首先通过免疫组织化学染色研究了136例肝癌和103个小DN（直径小于1厘米）中GPC3的表达情况，并利用X染色体失活嵌合体和雄激素受体（AR）基因多态性确定了81例女性患者来源DN的克隆性。然后，我们检测了这些样本在11个微卫星多态性位点的杂合性缺失（LOH）情况。结果显示，136例肝癌中有103例（75.7%）检测到GPC3免疫反应性，103个DN中有19例（18.4%）检测到，阳性率与HBsAg阳性相关。克隆性分析表明，15例女性患者来源的GPC3阳性DN（包括12例高级别DN，HGDN）以及66例GPC3阴性DN中的28例（42.4%）为单克隆。此外，在19例GPC3阳性DN中，在D6S1008位点（100%，19/19）、D8S262位点（52.6%，10/19）和D11S1301位点（57.9%，11/19）发现了染色体LOH。然而，GPC3阴性DN在三个位点的LOH频率分别为5.95%（5/84）、23.8%（20/84）和4.76%（4/84）。因此，我们得出结论，GPC3阳性DN，尤其是GPC3阳性HGDN，确实是肝癌的晚期癌前病变。
