转录因子Tex10募集p300乙酰化调节STAT3促进肝癌转移的机制研究

Tumor recurrence and metastasis are the main factors affecting the prognosis of hepatocellular carcinoma (HCC). Further study on the molecular mechanism of metastasis will provide an important basis for the diagnosis and treatment of HCC. Testis expressed 10 (Tex10) is a newly discovered pluripotency transcription factor. Our previous work indicated that Tex10 regulates the characteristics of liver cancer stem cells and promotes the formation of malignant phenotypes such as invasion and metastasis of HCC. Meanwhile, it promotes signal transducer and activator of transcription 3 (STAT3) acetylation, and protein immunoprecipitation analysis revealed that Tex10 could directly interact with histone acetyltransferase p300. Therefore, we speculated that Tex10 may promote HCC metastasis by recruiting p300 to activate the acetylation of STAT3. This project intends to further use various interventions to determine the molecular mechanism of Tex10 combined with p300 to acetylate STAT3. The effects of Tex10 on the biological behaviors of liver cancer stem cells formation, cell proliferation, apoptosis, invasion and metastasis were studied at the cellular level; Animal experiment revealed the promoting effect of Tex10 on HCC metastasis; the correlation between expression level of Tex10 and clinical pathological features such as HCC staging and prognosis was analyzed. This study contributes to elucidate the new mechanism of HCC metastasis and provide new targets and strategies for the diagnosis and treatment of HCC.

肿瘤复发和转移是影响肝癌预后的主要原因，对肝癌转移分子机制的深入研究将为肿瘤诊断和治疗提供重要依据。Tex10 是新发现的多能性转录因子，我们前期工作表明：Tex10调控肝癌干细胞特性，并促进肝癌细胞侵袭转移等恶性表型的形成，同时促进STAT3乙酰化，蛋白质免疫共沉淀分析发现Tex10与组蛋白乙酰转移酶p300相互结合。由此，我们推测：Tex10可能通过募集p300乙酰化激活STAT3促进肝癌转移。本项目拟进一步采用各种干预手段从分子水平确定 Tex10结合p300乙酰化激活STAT3的分子机制；从细胞水平研究 Tex10对肝癌干细胞形成、肝癌细胞增殖、凋亡、侵袭和转移等生物学行为的影响；动物水平揭示Tex10对肝癌转移的促进作用；临床标本分析Tex10表达水平与肝癌分期、预后等临床病理学特征的关系。本研究有助于进一步阐明肝癌转移的新机制，为诊断和治疗肝癌提供新靶点、新策略。

背景:睾丸表达蛋白10 (Testis expressed protein 10，Tex10)被报道与多种癌症的发生有关，其在肝细胞癌(hepatocellular carcinoma，HCC)转移中的作用尚未被研究。.方法:本研究采用western blot和免疫组化(IHC)分别检测Tex10在肝癌细胞系和组织芯片中的表达。通过RNA-Seq转录组分析鉴定Tex10介导的生物学过程。应用CCK-8、集落形成、transwell实验、裸鼠移植瘤生长和肺转移实验，评价Tex10对细胞增殖、迁移、侵袭和转移的体外和体内影响。通过免疫印迹、免疫组化、双荧光素酶报告、小干扰RNA (siRNA)、免疫共沉淀、免疫荧光和染色质免疫沉淀实验进一步研究其潜在机制。.结果:我们发现，与邻近正常组织相比，Tex10在HCC肿瘤组织中表达上调，其表达与不良预后相关。功能富集分析显示，Tex10敲低会改变许多生物学过程，尤其是细胞运动和细胞迁移。功能研究表明，Tex10可促进HCC细胞在体内外的增殖、迁移、侵袭和转移。此外，Tex10被证明通过STAT3信号通路调控细胞侵袭和EMT。在机制上，Tex10被发现与STAT3相互作用并促进其转录活性。此外，我们发现Tex10促进p300介导的STAT3乙酰化，而p300沉默则会抑制Tex10促进的STAT3转录活性。.结论:综上所述，这些研究结果表明Tex10通过上调STAT3活性发挥癌基因的作用，从而提示Tex10可能作为HCC患者预后的生物标志物和/或治疗靶点。