睾丸特异性蛋白1调控雄激素受体促进男性肝癌发生发展的分子机制研究

Among the hepatitis B virus (HBV) based hepatocellular carcinoma (HCC) patients, the majority are males. As the published report, HBV transcription can be enhanced by androgen receptor (AR). Our previous study have found that testis-specific protein Y-encoded 1 (TSPY1), which was encoded by gene from gonadoblastoma region on Y chromosome, was significantly associated with male HCC. Overexpression of TSPY1 in cancer tissues from HBV based male HCC patients was identified and validated. And the increasing ability of proliferation and invasion was observed and it was also confirmed that AR expression was up regulated simultaneously with TSPY1 overexpression in TSPY1 transfected HCC cell lines, but the molecular mechanism remains unclear. Accordingly, we put forward the hypothesis that TSPY1 might promote the tumorigenesis and development of male HCC via up regulating AR expression. By using the HCC cell lines with Y chromosome, liver tissue samples, neoplastic transformation model and xenograft HCC model in nude mice, we will investigate the effect of TSPY1 on the tumorigenesis and development of male HCC by the advanced techniques including gene transfection, RNA interference, tandem affinity purification combined with mass spectrometry identification, chromatin immune coprecipitation and signal pathway analysis. And our research will provide the evidences to explore the novel TSPY1 targeted method in treatment and prevention of male HCC.

乙型肝炎病毒（HBV）相关肝细胞癌患者男性多于女性，且雄激素受体（AR）可增强HBV转录促进肝癌发生。我们前期研究发现Y染色体基因编码的睾丸特异性蛋白1（TSPY1）与男性肝癌密切相关，并验证其在HBV背景的男性肝癌组织中高表达；过表达TSPY1可增强肝癌细胞的增殖和侵袭能力。预实验亦证实过表达TSPY1可上调肝癌细胞中AR的表达，但其具体机制有待探讨。据此提出假说：TSPY1通过上调AR的表达促进男性肝癌的发生发展。本项目拟以Y染色体阳性的肝癌细胞系、肝癌组织标本、肝癌致瘤性转化模型及裸鼠移植瘤模型为研究对象，采用基因转染、RNA干扰、串联亲和纯化结合质谱鉴定、染色质免疫共沉淀及信号通路研究等技术，从分子、细胞、组织以及动物等多个层次明确TSPY1在男性肝癌发生发展中的作用，阐明TSPY1调控AR表达的分子机制，探索以TSPY1为靶点的男性肝癌防治新思路。

乙型肝炎病毒（HBV）相关肝细胞癌患者男性多于女性，且雄激素受体（AR）可增强HBV转录促进肝癌发生。我们前期研究发现Y染色体基因编码的睾丸特异性蛋白1（TSPY1）与男性肝癌密切相关，并验证其在HBV背景的男性肝癌组织中高表达；过表达TSPY1可增强肝癌细胞的增殖和侵袭能力。预实验亦证实过表达TSPY1可上调肝癌细胞中AR的表达，但其具体机制有待探讨。据此提出假说：TSPY1通过上调AR的表达促进男性肝癌的发生发展。本项目首先明确了TSPY1在Y 染色体阳性的肝癌细胞和男性肝癌组织中高表达，且可以促进肝癌细胞的增殖，抑制凋亡。进一步研究发现，TSPY1和AR在肝癌细胞和组织中存在共定位，呈高度相关性表达，且过表达TSPY1可以上调AR的表达，而沉默TSPY1则减低AR的表达。免疫共沉淀结果提示TSPY1和AR存在相互作用。信号通路实验发现，TSPY1可活化MAPK/ERK信号通路，增强AR的表达，抑制MAPK/ERK信号通路后，AR表达减低。由此可知，TSPY1可通过MAPK/ERK信号通路来调控AR的表达。最后在临床组织样本中验证了TSPY1和AR表达存在高度相关性。本研究通过阐明TSPY1调控AR表达的分子机制，探索以TSPY1为靶点的男性肝癌防治新思路。