Department of Gastroenterology, China-Japan Friendship Hospital, 100029, Beijing, P. R. China.
Cancer Gene Ther. 2021 Aug;28(7-8):875-891. doi: 10.1038/s41417-020-0201-z. Epub 2020 Sep 26.
Hepatocellular carcinoma (HCC) is recognized as the most common malignancy of the liver in adults. Many human cancers have been associated with the oncogenic activation of the Wnt/β-catenin signaling pathway. The secreted frizzled-related proteins (sFRPs) function as negative regulators of the Wnt signaling and have important implications in carcinogenesis. This study aims to investigate the possible regulatory effects of sFRP3 on the Wnt/β-catenin signaling pathway and their interactions in HCC occurrence. Firstly, sFRP3 expression was quantified in the collected cancer and adjacent normal tissue samples from HCC patients. The lowly expressed sFRP3 in HCC tissues was found to be correlated with HCC development. The expression of sFRP3 was regulated by a lentivirus-based packaging system, and the Wnt/β-catenin signaling pathway was inactivated by DDK-1 in HepG2 cells. The expressions of Wnt1, β-catenin and the nuclear translocation of β-catenin were determined, both of which were down-regulated by sFRP3 overexpression. CCK8 assay, EdU staining, Colony formation assay, flow cytometry, scratch test and Transwell assay were employed to test cell viability, proliferation, cell cycle, apoptosis, migration and invasion, respectively. Overexpressed levels of sFRP3 were found to produce a reduction in MMP-2, MMP-7, MMP-9, PCNA, Ki67, and Bcl-2 expressions but an increase in the expressions of caspase-3 and Bax. In addition, overexpression of sFRP3 inhibited cell proliferation, migration, invasion, and colony formation, but promoted cell cycle arrest and cell apoptosis in HCC cells. The addition of the Wnt/β-catenin signaling pathway inhibitor, DKK-1, reversed the contributory effect of sFRP3 silencing on HCC development. Lastly, in vivo tumor formation was inhibited by enforced sFRP3 expressions. The obtained results suggested that sFRP3 acts as an anti-oncogene in HCC by inhibiting the activation of the Wnt/β-catenin signaling pathway.
肝细胞癌（HCC）是成人中最常见的肝脏恶性肿瘤。许多人类癌症与 Wnt/β-连环蛋白信号通路的致癌激活有关。分泌卷曲相关蛋白（sFRP）作为 Wnt 信号的负调节剂，在致癌作用中具有重要意义。本研究旨在探讨 sFRP3 对 HCC 发生中 Wnt/β-连环蛋白信号通路的可能调节作用及其相互作用。首先，定量检测了 HCC 患者癌组织和癌旁正常组织中 sFRP3 的表达。发现 HCC 组织中低表达的 sFRP3 与 HCC 的发生发展有关。通过基于慢病毒的包装系统调节 sFRP3 的表达，并在 HepG2 细胞中用 DDK-1 使 Wnt/β-连环蛋白信号通路失活。测定 Wnt1、β-连环蛋白和β-连环蛋白的核转位，结果表明 sFRP3 过表达均可下调其表达。CCK8 测定、EdU 染色、集落形成测定、流式细胞术、划痕试验和 Transwell 测定分别用于测试细胞活力、增殖、细胞周期、凋亡、迁移和侵袭。结果发现，过表达 sFRP3 可降低 MMP-2、MMP-7、MMP-9、PCNA、Ki67 和 Bcl-2 的表达水平，同时增加 caspase-3 和 Bax 的表达水平。此外，过表达 sFRP3 可抑制 HCC 细胞的增殖、迁移、侵袭和集落形成，但促进细胞周期停滞和细胞凋亡。添加 Wnt/β-连环蛋白信号通路抑制剂 DKK-1 可逆转 sFRP3 沉默对 HCC 发展的促进作用。最后，强制表达 sFRP3 抑制了体内肿瘤的形成。结果表明，sFRP3 通过抑制 Wnt/β-连环蛋白信号通路的激活在 HCC 中发挥抑癌基因的作用。
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