目的：探讨TRIM21 通过激活Wnt/β-catenin 信号通路调控卵巢癌细胞增殖以及PARP抑制剂耐药的相关机制。方法：收集2018 年1 月至2019 年1 月于延安市人民医院手术切除的8 例卵巢癌组织及宫颈上皮组织标本，并根据患者是否对PARP抑制剂尼拉帕尼耐药分为耐药组和非耐药组（4 例/组）；卵巢癌细胞系CAOV3、SKOV3、OVCAR3、ES-2、HO8910、A2780 和OV2008。用qPCR 法和Western blotting（WB）分别检测卵巢癌组织和细胞系中TRIM21 和β-catenin 表达水平。构建TRIM21 过表达及敲减细胞系，用CCK-8 法检测各组细胞的增殖活力，用TOP/FOP 双荧光素酶实验检测TRIM21 对Wnt信号通路激活的影响，qPCR法和WB检测TRIM21 对β-catenin mRNA和蛋白水平的影响，并通过Wnt通路抑制剂XAV-939 作进一步验证。结果：TRIM21 在卵巢癌组织中的表达水平显著高于宫颈上皮组织（P<0.01），且在耐药组织中表达水平较高（P<0.01）；TRIM21 在卵巢癌ES-2 细胞中表达水平相对最高，而在CAOV3 和A2780 中表达水平相对较低（均P<0.01）。敲减TRIM21 后，ES-2 细胞中TRIM21 的表达水平显著降低（P<0.01），细胞的增殖能力明显降低（P<0.01）；过表达TRIM21 后，CAOV3 细胞的增殖能力显著增强（P<0.01），并可抑制尼拉帕尼的抗肿瘤增殖活性，其可调控Wnt/catenin 通路的激活，而敲减β-catenin 或使用Wnt/β-catenin 抑制剂XAV-939 后，TRIM21 在卵巢癌中的作用被显著逆转。结论：TRIM21 可通过调控Wnt/β-catenin 信号通路增强卵巢癌细胞的增殖能力并在PARP抑制剂尼拉帕尼耐药过程中发挥一定的作用。

Objective: To explore the mechanism of TRIM21 regulating the proliferation of ovarian cancer cells and the resistance of PARP inhibitors by activating Wnt/β-catenin signaling pathway. Methods: Eight pairs of ovarian cancer tissues and cervical epithelial tissues that surgically removed at Yan'an People's Hospital from January 2018 to January 2019 were collected for this study. And the tissues were classified into resistant group and non-resistant group (4 case/group) according to whether the patients were resistant to PARP inhibitor (nilapanib). Ovarian cancer cell lines CAOV3, SKOV3, OVCAR3, ES-2, HO8910, A2780 and OV2008 were also collected for this study. qPCR and Western blotting (WB) were used to detect the expression levels of TRIM21 and β -catenin in the above mentioned tissues and cell lines. Cell lines with TRIM21 overexpression and knockdown were constructed. CCK-8 method was used to detect the proliferation activity of ovarian cancer cells in each group, TOP/FOP dual luciferase assay was used to detect the effect of TRIM21 on Wnt signaling pathway activation, qPCR and WB were used to detect the effect of TRIM21 on mRNA and protein levels of β-catenin, which was further verified by Wnt pathway inhibitor XAV-939. Results: The expression level of TRIM21 in ovarian cancer tissues was significantly higher than that in cervical epithelial tissues (P<0.01), and its expression was more higher in the drug-resistant tissues (P<0.01). TRIM21 expression was the highest in ES-2 cells but comparatively low in CAOV3 and A270 cells (all P<0.01). After TRIM21 knockdown, the expression of TRIM21 in ES-2 cells was significantly decreased, and the cell proliferation was significantly reduced (all P<0.01). After overexpressing TRIM21, the proliferative capacity of ovarian cancer CAOV3 cells was significantly increased (P<0.01), and the antitumor effect of nilaparib was inhibited; TRIM21 overexpression could regulate Wnt/β-catenin pathway activation, while β -catenin knockdown or Wnt/β-catenin inhibitor XAV-939 could significantly reverse the effect of TRIM21 in ovarian cancer. Conclusion: TRIM21 can enhance the proliferation of ovarian cancer cells via regulating Wnt/β-catenin pathway, it plays a certain role in the process of drug resistance of PARP inhibitor nilapani.

陕西省教育厅自然科学研究项目资助（No. 2013JK0782）