Zhao Xiaoli, Lei Yutian, Zhu Han, Shen Wenyi, Qian Sixuan, Li Jianyong, Zhu Yu
Department of Hematology, The First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Key Laboratory of Hematology of Nanjing Medical University, Guangzhou Road 300, Nanjing, 210029, China.
J Transl Med. 2025 May 26;23(1):585. doi: 10.1186/s12967-025-06429-y.
Due to high heterogeneity, diagnosing MDS can be challenging. Consequently, investigating its pathogenesis and progression mechanisms, and seeking novel targets for diagnosis and treatment, are critical issues that require urgent attention. This study aimed to investigate whether LRRFIP1 might contribute to MDS pathogenesis by modulating the Wnt/β-catenin signaling pathway.
In MDS cell lines, mRNA transcriptome sequencing and Dual luciferase reporter gene assays were employed to assess Wnt/β-catenin signaling pathway activity. To explore the biological characteristics of MDS cell lines, CCK8, Annexin-V APC/7-AAD and Annexin V-FITC/PI double staining and fluorescence TUNEL assay, and PI single staining were used.
In MDS cell lines, proliferation was notably higher in LRRFIP1-overexpressing cells compared to silenced ones, while cell apoptosis was lower in the former. mRNA transcriptome sequencing revealed LRRFIP1's involvement in modulating the Wnt/β-catenin signaling pathway. LRRFIP1 was found to positively regulate Wnt/β-catenin pathway activity, and synergy between LRRFIP1 and Dvl was observed in enhancing canonical Wnt signaling. LRRFIP1 overexpression significantly upregulated key genes in Wnt signaling pathway (such as β-catenin, Dvl2, Dvl3 and Wnt) at the mRNA level, and notably upregulated non-phosphorylated β-catenin at the protein level. Moreover, BCL-2 and CyclinD1 protein expression was significantly higher in LRRFIP1-overexpressing cells compared to silenced ones, with even greater expression observed in LRRFIP1/Dvl3 co-overexpressing cells.
LRRFIP1 can promote the proliferation of MDS cells and inhibit apoptosis, and LRRFIP1 and Dvl can synergistically enhance the activity of the Wnt/β-catenin signaling pathway in MDS, thus providing evidence for LRRFIP1's involvement in the pathogenesis of MDS.
由于高度异质性，骨髓增生异常综合征（MDS）的诊断具有挑战性。因此，研究其发病机制和进展机制，并寻找新的诊断和治疗靶点，是亟待关注的关键问题。本研究旨在探讨富含亮氨酸重复序列蛋白1（LRRFIP1）是否通过调节Wnt/β-连环蛋白信号通路促进MDS的发病机制。
在MDS细胞系中，采用mRNA转录组测序和双荧光素酶报告基因检测来评估Wnt/β-连环蛋白信号通路活性。为探究MDS细胞系的生物学特性，使用了CCK8、膜联蛋白V-别藻青蛋白/7-氨基放线菌素D和膜联蛋白V-异硫氰酸荧光素/碘化丙啶双重染色及荧光TUNEL检测，以及碘化丙啶单染色。
在MDS细胞系中，与LRRFIP1沉默的细胞相比，过表达LRRFIP1的细胞增殖明显更高，而前者的细胞凋亡更低。mRNA转录组测序显示LRRFIP1参与调节Wnt/β-连环蛋白信号通路。发现LRRFIP1正向调节Wnt/β-连环蛋白通路活性，并且在增强经典Wnt信号方面观察到LRRFIP1与Dishevelled（Dvl）之间存在协同作用。LRRFIP1过表达在mRNA水平显著上调Wnt信号通路中的关键基因（如β-连环蛋白、Dvl2、Dvl3和Wnt），并且在蛋白水平显著上调非磷酸化β-连环蛋白。此外，与LRRFIP沉默的细胞相比，过表达LRRFIP1的细胞中BCL-2和细胞周期蛋白D1蛋白表达显著更高，在LRRFIP1/Dvl3共过表达的细胞中观察到更高的表达。
LRRFIP1可促进MDS细胞增殖并抑制凋亡，并且LRRFIP1和Dvl可协同增强MDS中Wnt/β-连环蛋白信号通路的活性，从而为LRRFIP1参与MDS的发病机制提供证据。
