治疗诱导肿瘤分泌蛋白质组（TIS）介导的肝细胞癌对Sorafenib耐受的机制研究

The natural and acquired resistance against Sorafenib in hepatocellular carcinoma (HCC) is the main obstacle that limits its efficiency in patients diagnosed at advanced stages. The latest theory is that therapy-induced tumor secrectomes (TIS) which is transcriptionally regulated in sensitive tumor cells activates the downstream AKT signaling, promoting the survival of sensitive cells and proliferation and invasion of resistant cells. Our preliminary research revealed the Sorafenib could activate AKT in HCC cell which was capable of inducing drug resistance. In further experiments, we found that the incubation of Sorafenib with sensitive HCC cells and resistant HCC cell could promote the proliferation, migration, invasion of resistant HCC cell, and also promote tumor metastasis in nude mice lung. So we speculated that TIS which could be responsible for AKT activation might be involved in Sorafenib resistance. On this basis, we aim to investigate the following problems in the level of molecular, cell and animal: Whether Sorafenib induces TIS in HCC? Whether TIS was transcriptionally regulated? What is the downstream signaling responsible for TIS induced Sorafenib resistance? Whether the combination of Sorafenib and the related signaling inhibitor could enhance Sorafenib efficiency? This study is expected to reveal a novel mechanism underlying Sorafenib resistance which will provide new molecular targets and theoretical basis to improve Sorafenib efficiency and reduce HCC progression.

肝细胞癌(HCC)存在对Sorafenib的耐受是制约其在晚期HCC患者中疗效的主要原因。最新理论认为，敏感细胞产生的治疗诱导肿瘤蛋白质组(TIS)通过激活AKT通路，促进自身存活和耐药细胞增殖浸润，导致激酶抑制剂耐受。我们初步研究表明， Sorafenib作用于HCC细胞可出现AKT上调，促进肿瘤耐药。在敏感细胞和耐受细胞共培养体系中，Sorafenib可促进耐受细胞的增殖、侵袭和转移灶形成，提示Sorafenib作用于敏感HCC细胞可能产生TIS。在此基础上本项目拟明确：Sorafenib作用于HCC后是否产生TIS；TIS是否由基因转录水平调控；TIS的下游信号通路是什么； Sorafenib联合TIS下游信号通路抑制剂是否可逆转Sorafenib耐药。本项目有望以全新理论解释HCC对Sorafenib的耐受和肿瘤进展机制，为提高Sorafenib疗效、延缓肿瘤进展提供新的治疗策略。

肝细胞癌(HCC)存在对Sorafenib的耐受是制约其在晚期HCC患者中疗效的主要原因。有研究显示，敏感细胞产生的治疗诱导肿瘤蛋白质组(TIS)通过激活AKT通路，促进自身存活和耐药细胞增殖浸润，导致激酶抑制剂耐受。Regorafenib是最新经FDA批准的用于Sorafenib治疗后肿瘤进展HCC患者的二线用药，但其抑制Sorafenib耐受的机制及其自身耐受的机制未有报道。本项目主要开展了TIS介导的HCC对Sorafenib耐受的机制研究，在研究过程中，又陆续进行了进展期HCC的二线靶向治疗用药Regorafenib逆转Sorafenib耐受的机制研究，Regorafenib逆转HGF诱导的Sorafenib耐受的机制研究，以及Regorafenib耐药机制的研究。我们的研究发现低剂量Sorafenib作用于敏感HCC细胞后获得的条件培养基可促进敏感HCC细胞增殖，抑制细胞凋亡，促进耐受细胞增殖和侵袭迁移。条件培养基可通过激活AKT通路促进肿瘤恶性行为，而联合应用AKT抑制剂可逆转HCC细胞增殖和侵袭迁移。同时，我们的研究还发现，Regorafenib可通过抑制HGF/MET/ERK和HGF/MET/STAT3通路逆转Sorafenib耐受。此外，我们发现低剂量长时间刺激可诱导HCC产生对Regorafenib耐受，耐受细胞中出现HGF通路活化，联合应用HGF通路抑制剂Crizotinib可逆转Regorafenib耐受。我们的研究揭示了进展期HCC一线靶向治疗用药Sorafenib的耐受机制，Sorafenib治疗失败后二线用药Regorafenib逆转Sorafenib耐受的机制，以及Regorafenib的耐受机制，较为系统和全面地阐述了肝癌靶向治疗耐受的机制，为提高抗肝癌疗效、延缓肿瘤进展提供新的治疗策略和理论基础，具有潜在的临床应用价值。