[摘 要] 目的：探索lncRNA FOXD2-AS1通过EZH2调控LATS1表达影响肾透明细胞癌（ccRCC）细胞增殖与迁移的作用机 理。方法：采用GEPIA 2软件在线分析TCGA数据库中FOXD2-AS1在ccRCC组织中的表达水平，评估其与患者总体生存率之 间的关系。用qPCR法检测肾癌细胞及临床收集的26例ccRCC组织中FOXD2-AS1表达水平，用CCK-8法和Transwell小室实验 观察敲减 FOXD2-AS1 表达对肾癌细胞增殖与迁移的影响；用 qPCR 与 WB 法检测敲减 FOXD2-AS1 表达对肾癌细胞 LATS1 mRNA与蛋白表达的影响。用RNA免疫沉淀（RIP）和染色质免疫沉淀（ChIP）法分析FOXD2-AS1与EZH2及LATS1之间的作 用。结果：GEPIA 2软件分析结果显示，FOXD2-AS1在ccRCC组织中呈明显高表达（P < 0.01），FOXD2-AS1高表达患者的总体 生存率较低（P < 0.05）。qPCR法检测结果显示，相较癌旁组织，FOXD2-AS1在26例ccRCC组织中呈显著高表达（P < 0.01）。相 较永生化肾小管上皮细胞HK-2，FOXD2-AS1 在肾癌 786-O、ACHN 及 SN12-PM6 细胞中均显著高表达（均 P < 0.01）。敲减 FOXD2-AS1表达，均可显著降低肾癌细胞的增殖与迁移能力（均P < 0.05），并明显上调LATS1的mRNA与蛋白表达水平（均 P < 0.01）。RIP 与 ChIP 实验证实，FOXD2-AS1 可结合并招募 EZH2 至 LATS1 启动子区域而发挥作用。挽救实验显示，敲减 LATS1或过表达EZH2 可部分逆转敲减 FOXD2-AS1 对肾癌细胞增殖与迁移的抑制作用。结论：FOXD2-AS1在ccRCC中 高表达，其通过募集EZH2至LATS1基因启动子区域而负性调控LATS1的表达，进而促进肾癌细胞的增殖与迁移。

[Abstract] Objective: To explore the mechanism of lncRNA FOXD2-AS1 regulating the expression of LATS1 via EZH2 to affect the proliferation and migration of clear cell renal cell carcinoma (ccRCC) cells. Methods: The GEPIA 2 online tool was used to analyze the expression levels of FOXD2-AS1 in ccRCC tissues from the Cancer Genome Atlas (TCGA) database, and their correlation with patients’ overall survival rates was evaluated. Quantitative PCR (qPCR) was performed to analyze the expressions of FOXD2-AS1 in renal cancer cells and 26 clinically collected ccRCC tissue samples. CCK-8 cell proliferation assay and transwell chamber migration assay were employed to observe the effects of FOXD2-AS1 knockdown on the proliferation and migration of renal cancer cells. qPCR and Western blot analysis were utilized to assess the impact of FOXD2-AS1 knockdown on the expression of LATS1. RNA immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP) assays were performed to analyze the interaction between FOXD2-AS1, EZH2, and LATS1. Results: The GEPIA 2 software analysis revealed that FOXD2-AS1 was significantly upregulated in ccRCC tissues (P < 0.01) and patients with high FOXD2-AS1 expression exhibited lower overall survival rates (P < 0.05). The qPCR analysis results showed that FOXD2-AS1 was significantly upregulated in 26 samples of ccRCC tissues compared with adjacent normal kidney tissues (P < 0.01). Compared with immortalized renal tubular epithelial cell line HK-2, the expression of FOXD2-AS1was significantly elevated in three types of renal cancer cell lines (786-O, ACHN and SN12-PM6) (P < 0.01). Knockdown of FOXD2-AS1 expression significantly decreased the proliferation and migration abilities of renal cancer cells (P < 0.05), and markedly increased the mRNA and protein expression levels of LATS1 (all P < 0.01). RIP and ChIP assays confirmed that FOXD2-AS1 can bind and recruit EZH2 to the promoter region of LATS1 to exert its effect. Salvage experiments demonstrated that knocking down LATS1 or overexpressing EZH2 partially reversed the inhibitory effect of FOXD2-AS1 knockdown on the proliferation and migration of renal cancer cells. Conclusion: FOXD2-AS1 is highly expressed in ccRCC, and it negatively regulates the expression of LATS1 by recruiting EZH2 to the promoter region of the LATS1 gene, thereby facilitating the proliferation and migration of renal cancer cells.

国家自然科学基金项目（No. 81502204；No.82002708）