ARHGAP10调控细胞骨架信号通路与卵巢癌侵袭、转移的相关性研究

Ovarian cancer is the most common and lethal gynecological cancer. ARHGAP10 gene was chosen as the cut point of this study, and it might be an important tumor suppressor gene in ovarian cancer. GSEA/KEGG analysis found that ARHGAP10 is closely related to cytoskeleton pathway，STRING analysis showed the functional interaction between ARHGAP10 and Rho family. This project attempts to explore the role of ARHGAP10 in the pathogenesis of ovarian cancer, RNA interference technology will be adopted to overexpression and knockdown ARHGAP10 gene in A2780 and OVCAR3 cell lines, PCR-array analysis the gene expression of cytoskeleton and EMT signal pathway. Identification of the key genes regulated by ARHGAP10, as well as the regulation effect of ARHGAP10 on the cytoskeleton and EMT signaling pathway. At the same time, we will verified the differential expression of key genes which regulated by ARHGAP10 in the 50 cases of serous ovarian carcinoma specimens (primary tumor, metastasis, recurrence), relationship between ARHGAP10/key gene and the invasion, metastasis and recurrence of ovarian cancer. At last, we believe that our results may provide a new theoretical basis of gene diagnosis, treatment, and prognosis evaluation of ovarian cancer.

卵巢癌是妇科最常见的致死性肿瘤。前期研究我们选择ARHGAP10基因为切入点。证实ARHGAP10可能是卵巢癌重要的抑癌基因。进一步GSEA/KEGG分析，发现ARHGAP10与细胞骨架信号通路密切相关，STRING分析显示ARHGAP10与Rho家族存在功能相互作用；为了细析ARHGAP10致瘤机制，本课题建立抑制/过表达ARHGAP10卵巢癌细胞系OVCAR3及A2780，采用PCR-array技术，检测分析细胞骨架和EMT通路信号基因表达；筛选和验证ARHGAP10调控的关键基因，探究ARHGAP10调控细胞骨架和EMT信号通路模式。同时，选择50例完整系列的卵巢浆液性癌组织标本（原发灶、转移灶、复发灶），验证ARHGAP10及其调控的关键基因的表达差异，建立ARHGAP10及其关键基因与卵巢癌侵袭、转移和复发的相关性。课题研究可为今后卵巢癌基因诊断治疗及预后评估提供新的理论依据。

卵巢癌是妇科最常见的致死性肿瘤，寻找肿瘤发生发展、转移及复发过程中的特异基因，并注释基因功能，研究相关信号通路，可为肿瘤的治疗提供新的理论依据。前期研究我们选择ARHGAP10基因为切入点，证实ARHGAP10可能是卵巢癌重要的抑癌基因。我们研究发现， ARHGAP10 在卵巢癌组织中的表达明显低于癌旁组织，与 TCGA 数据库资料相符，并且与患者的总生存期呈负相关。体外实验证实，ARHGAP10 过表达能抑制细胞增殖能力，能在 G1 期抑制细胞增殖并促进细胞凋亡； ARHGAP10 过表达还能及抑制细胞黏附、转移及侵袭能力。小鼠皮下成瘤实验也证实， ARHGAP10 过表达能抑制肿瘤的生长。此外，我们进一步证实，ARHGAP10 与 Cdc42 存在相互作用，ARHGAP10 能抑制 Cdc42 的活性进而抑制细胞的增殖、侵袭、转移及黏附能力。CXCL12/CXCR4是介导卵巢癌侵袭和转移的重要信号通路之一。我们进一步研究发现，CXCL12/CXCR4通过抑制ARHGAP10的表达进而促进卵巢癌细胞的侵袭。我们证实，卵巢癌及癌旁组织中ARHGAP10与CXCL12、CXCR4、VEGF及VEGFR2的表达水平呈负相关；CXCL12能抑制卵巢癌细胞ARHGAP10的表达，而CXCR4 抑制剂AMD3100或VEGFR2抑制剂SU1498则能抑制CXCL12介导的ARHGAP10降表达；过表达ARHGAP10则能够抑制CXCL12介导的促卵巢癌细胞侵袭作用。体外实验证实，敲降ARHGAP10表达能显著促进卵巢癌细胞肺转移，同时能够抑制SU1498的抑制卵巢癌细胞肺转移作用。本研究提示：ARHGAP10可能作为一种重要的抑癌基因，通过调控 Cdc42 的活性直接参与调控卵巢癌细胞的生长、分化、转移；且ARHGAP10 的调控机制可能涉及作为调控卵巢癌细胞侵袭和转移的CXCL12/CXCR4信号通路的下游关键分子；因此，ARHGAP10可作为潜在的治疗卵巢癌的关键靶点。