卵巢癌是最常见的妇科恶性肿瘤之一，预后较差。 THUMPD3-AS1 是多种癌症中的致癌长链非编码 RNA (lncRNA)。此外，miR-320d在卵巢癌细胞中下调并抑制增殖，而ARF1在上皮性卵巢癌中上调并促进恶性进展。然而，THUMPD3-AS1在卵巢癌中的作用及其潜在机制尚未阐明。采用人正常卵巢上皮细胞（IOSE80）和卵巢癌细胞系（CAVO3、A2780、SKOV3、OVCAR3 和 HEY）进行体外实验。使用 CCK-8、流式细胞术和 TUNEL 测定法确定 THUMPD3-AS1 在细胞活力和细胞凋亡中的功能作用。进行蛋白质印迹以评估 ARF1、Bax、Bcl-2 和 caspase 3 的蛋白水平，而 RT-qPCR 则用于测量 ARF1 mRNA、THUMPD3-AS1 和 miR-320d 水平。通过双荧光素酶测定验证了 miR-320d 与 THUMPD3-AS1 或 ARF1 之间的靶向关系。 THUMPD3-AS1和ARF1在卵巢癌细胞中高表达，而miR-320d水平低表达。 THUMPD3-AS1 敲低能够抑制细胞活力并加速 OVCAR3 和 SKOV3 细胞的凋亡。此外，THUMPD3-AS1 充当 miR-320d 的海绵，防止 ARF1 的降解。 MiR-320d 下调逆转了 THUMPD3-AS1 耗竭诱导的肿瘤抑制功能。此外，miR-320d 过表达会抑制卵巢癌细胞活力并加速细胞凋亡，而 ARF1 过表达则可逆转这种情况。 THUMPD3-AS1 通过调节 miR-320d/ARF1 轴抑制卵巢癌细胞凋亡。这些发现可能为卵巢癌治疗提供前瞻性目标。© 2024 美国实验生物学会联合会。

Ovarian cancer is one of the most common gynecologic malignancies that has a poor prognosis. THUMPD3-AS1 is an oncogenic long noncoding RNA (lncRNA) in several cancers. Moreover, miR-320d is downregulated and inhibited proliferation in ovarian cancer cells, whereas ARF1 was upregulated and promoted the malignant progression in epithelial ovarian cancer. Nevertheless, the role of THUMPD3-AS1 in ovarian cancer and the underlying mechanism has yet to be elucidated. Human normal ovarian epithelial cells (IOSE80) and ovarian cancer cell lines (CAVO3, A2780, SKOV3, OVCAR3, and HEY) were adopted for in vitro experiments. The functional roles of THUMPD3-AS1 in cell viability and apoptosis were determined using CCK-8, flow cytometry, and TUNEL assays. Western blot was performed to assess the protein levels of ARF1, Bax, Bcl-2, and caspase 3, whereas RT-qPCR was applied to measure ARF1 mRNA, THUMPD3-AS1, and miR-320d levels. The targeting relationship between miR-320d and THUMPD3-AS1 or ARF1 was validated with dual luciferase assay. THUMPD3-AS1 and ARF1 were highly expressed in ovarian cancer cells, whereas miR-320d level was lowly expressed. THUMPD3-AS1 knockdown was able to repress cell viability and accelerate apoptosis of OVCAR3 and SKOV3 cells. Also, THUMPD3-AS1 acted as a sponge of miR-320d, preventing the degradation of ARF1. MiR-320d downregulation reversed the tumor suppressive function induced by THUMPD3-AS1 depletion. Additionally, miR-320d overexpression inhibited ovarian cancer cell viability and accelerated apoptosis, which was overturned by overexpression of ARF1. THUMPD3-AS1 inhibited ovarian cancer cell apoptosis by modulation of miR-320d/ARF1 axis. The discoveries might provide a prospective target for ovarian cancer treatment.© 2024 Federation of American Societies for Experimental Biology.