Department of Hematology and Oncology, Shenzhen Children's Hospital, Shenzhen, Guangdong, China (mainland).
Med Sci Monit. 2020 Jun 29;26:e923799. doi: 10.12659/MSM.923799.
BACKGROUND Upregulation of the Wnt/ß-catenin pathway has been demonstrated to promote tumor proliferation and chemoresistance in lymphoma. Our objective was to evaluate the effect of the Wnt/ß-catenin pathway inhibitor WNT974 in lymphoma cells. MATERIAL AND METHODS Human lymphoma cell lines HUT-78 and BJAB were treated with or without 1 μM WNT974±0.15 μg/L doxorubicin (Dox). Cell viability and proliferation were evaluated by CCK-8 and colony formation assay. Expression of proliferating cell nuclear antigen (PCNA), KI67, and apoptotic-related proteins including Bcl-2, Bax, cleaved-caspase3, and cleaved-caspase9, together with Wnt pathway proteins Wnt, ß-catenin, Axin2, and c-Myc, were detected by Western blot analysis. Flow cytometry was used to calculate the ratio of apoptotic cells. RESULTS In HUT-78 and BJAB cells, 1 μM WNT974 significantly reduced viability and colony formation. The expression of 2 markers of tumor cell proliferation, protein PCNA and KI67, was also reduced by WNT974. Treatment with 1 μM WNT974 for 48 h increased the rate of cell apoptosis, inhibited the expression of anti-apoptotic protein Bcl-2, and enhanced pro-apoptotic proteins Bax, cleaved-caspase3, and cleaved-caspase9 expression in both cell lines. After treatment with WNT974 plus Dox, cell viability was markedly decreased compared with Dox treatment alone. Mechanistically, WNT974 prevented the expression of Wnt, Axin2, ß-catenin, and its target gene c-Myc. CONCLUSIONS WNT974 effectively treats lymphoma by inhibiting cell proliferation, inducing cell apoptosis, and enhancing chemosensitivity to Dox, and these effects are dependent on blocking Wnt/ß-catenin signaling.
Wnt/β-catenin 通路的上调已被证明可促进淋巴瘤中的肿瘤增殖和化疗耐药。我们的目的是评估 Wnt/β-catenin 通路抑制剂 WNT974 在淋巴瘤细胞中的作用。
用或不用 1 μM WNT974±0.15 μg/L 阿霉素（Dox）处理人淋巴瘤细胞系 HUT-78 和 BJAB。通过 CCK-8 和集落形成实验评估细胞活力和增殖。通过 Western blot 分析检测增殖细胞核抗原（PCNA）、KI67 和凋亡相关蛋白（包括 Bcl-2、Bax、cleaved-caspase3 和 cleaved-caspase9）以及 Wnt 通路蛋白 Wnt、β-catenin、Axin2 和 c-Myc 的表达。流式细胞术用于计算凋亡细胞的比例。
在 HUT-78 和 BJAB 细胞中，1 μM WNT974 显著降低细胞活力和集落形成。肿瘤细胞增殖的 2 个标志物，蛋白 PCNA 和 KI67 的表达也被 WNT974 降低。用 1 μM WNT974 处理 48 h 可增加细胞凋亡率，抑制抗凋亡蛋白 Bcl-2 的表达，并增强两种细胞系中促凋亡蛋白 Bax、cleaved-caspase3 和 cleaved-caspase9 的表达。与单独用 Dox 处理相比，用 WNT974 加 Dox 处理后细胞活力明显降低。从机制上讲，WNT974 通过阻止 Wnt、Axin2、β-catenin 和其靶基因 c-Myc 的表达来治疗淋巴瘤。
WNT974 通过抑制细胞增殖、诱导细胞凋亡和增强对 Dox 的化疗敏感性来有效治疗淋巴瘤，这些作用依赖于阻断 Wnt/β-catenin 信号通路。
