背景:据报道,抑制岩藻糖基转移酶4(FUT4)激活叉头盒O1(FOXO1)可导致癌细胞凋亡,然而,骨肉瘤的机制尚不清楚。
目的:探讨FUT4和FOXO1在骨肉瘤生长中的生物学意义。
方法:使用人成骨细胞系和骨肉瘤细胞系进行体外试验。使用QRT-PCR测定以及蛋白质印迹测定来确定细胞中FUT4和FOXO1的相对表达水平。通过使用CCK-8测定,菌落试验,EDU检测,伤口愈合试验和Transwell试验,骨肉瘤细胞增殖能力,迁移和入侵与si-FUT4的关系进行了检查。TUNEL试验用于评估Si对骨肉瘤细胞KHOS和U2OS凋亡的影响。用Westernblot法鉴定增殖细胞的表达,骨肉瘤细胞KHOS和U2OS的迁移和凋亡相关蛋白标记以及Wnt/β-catenin信号通路中重要蛋白的表达。
结果:与成骨细胞相比,骨肉瘤细胞表达更多的FUT4。骨肉瘤细胞增殖能力,入侵,和迁移被FUT4表达的抑制显著抑制,这也增加了骨肉瘤细胞的凋亡。通过上调FOXO1表达阻断Wnt/β-Catenin信号通路,其又通过抑制FUT4表达而被抑制。
结论:骨肉瘤细胞表达更多的FUT4。Wnt/β-catenin信号通路对骨肉瘤细胞死亡有显著影响,和抑制FUT4的表达可能靶向FOXO1激活,以降低骨肉瘤细胞的增殖能力,入侵,和迁移。
BACKGROUND: It has been reported that inhibition of Fucosyltransferase4 (FUT4) to activate Forkhead box O1 (FOXO1) can lead to apoptosis of cancer cells, however, the mechanism in osteosarcoma is still unclear.
OBJECTIVE: To explore the biological significance of the connection between FUT4 and FOXO1 in osteosarcoma growth.
METHODS: In vitro tests were conducted using the human osteoblast cell line and the osteosarcoma cell lines. QRT-PCR assay as well as western blot assay were used to ascertain the relative expression levels of FUT4 and FOXO1 in the cells. By using the CCK-8 assay, colony assay, EDU assay, wound healing assay and Transwell assay, osteosarcoma cells\' ability to proliferate, migrate and invade were examined in relation to si- FUT4. TUNEL test was used to evaluate Si-impact FUT4\'s on KHOS and U2OS apoptosis in osteosarcoma cells. Western blot assay was used to identify the expression of proliferative, migrating and apoptosis-related protein markers in osteosarcoma cells KHOS and U2OS and the expression of important proteins in the Wnt/ β-catenin signaling pathway.
RESULTS: In comparison with osteoblasts, osteosarcoma cells expressed more FUT4. The osteosarcoma cells\' capacities to proliferate, invade, and migrate were markedly inhibited by the inhibition of FUT4 expression, which also increased osteosarcoma cell apoptosis. The Wnt/β-catenin signaling pathway was blocked by upregulating FOXO1 expression, which was in turn inhibited by inhibiting FUT4 expression.
CONCLUSIONS: Osteosarcoma cells express more FUT4. The Wnt/β-catenin signaling pathway has a significant effect on osteosarcoma cell death, and inhibition of FUT4 expression may target FOXO1 activation to decrease osteosarcoma cells\' ability to proliferate, invade, and migrate.