Kong Zhenzhen, Zhou Ping, Xu Jiahao, Zhang Ying, Wang Yong
Department of Laboratory, Wujin Hospital Affiliated With Jiangsu University, No. 2 of Yongning North Road, Changzhou, 213002, Jiangsu, People's Republic of China.
The Wujin Clinical College of Xuzhou Medical University, Xuzhou, 221006, Jiangsu, People's Republic of China.
Cell Div. 2025 Mar 11;20(1):7. doi: 10.1186/s13008-025-00147-z.
Regulatory Factor X (RFX) transcription factors have been implicated in different cancers. Ras association domain family (RASSF) has been shown clinical significance in lung cancer. This paper was to investigate the interaction of RFX2 and RASSF1 in lung adenocarcinoma (LUAD).
The transcriptome differences of LUAD patients in GSE32863, GSE43458, and GSE21933 datasets were analyzed. A-549 and NCI-H358 cell lines after overexpression of RFX2 were co-cultured with activated CD8 T cells, and the release of IFN-γ, GZMB, PRF1 by CD8 T cells, and PD-L1 in the LUAD cells were detected. Cell viability, invasion, and apoptosis were analyzed by CCK-8, Transwell, and TUNEL assays. Dual-luciferase assay and ChIP were conducted to detect the interaction between RFX2 and RASSF1 promoter. An in vivo tumor model was constructed to monitor tumor growth. YAP protein levels and phosphorylation were measured. A-549 and NCI-H358 cells treated with DMSO or PY-60 after RFX2 overexpression were co-cultured with activated CD8 T cells.
RFX2 was notably downregulated in LUAD. RFX2 overexpression increased infiltrating CD8 T cells within transplanted tumors and inhibited immune escape, proliferation, and invasion of LUAD cells. RFX2 was enriched in the RASSF1 promoter, and RFX2 activated RASSF1 transcription by binding to the RASSF1 promoter. RASSF1 knockdown reversed the ability of RFX2 overexpression to inhibit immune escape. RFX2 depletion downregulated RASSF1, which reduced YAP phosphorylation, thus affecting the Hippo pathway to promote the immune escape.
RFX2 Loss in LUAD downregulates RASSF1 expression and YAP phosphorylation, thereby promoting immune escape through Hippo signaling.
调节因子X（RFX）转录因子与不同癌症有关。Ras关联结构域家族（RASSF）已在肺癌中显示出临床意义。本文旨在研究RFX2与RASSF1在肺腺癌（LUAD）中的相互作用。
分析GSE32863、GSE43458和GSE21933数据集中LUAD患者的转录组差异。将过表达RFX2后的A-549和NCI-H358细胞系与活化的CD8 T细胞共培养，检测CD8 T细胞释放的IFN-γ、GZMB、PRF1以及LUAD细胞中的PD-L1。通过CCK-8、Transwell和TUNEL实验分析细胞活力、侵袭和凋亡情况。进行双荧光素酶实验和染色质免疫沉淀实验以检测RFX2与RASSF1启动子之间的相互作用。构建体内肿瘤模型以监测肿瘤生长。检测YAP蛋白水平和磷酸化情况。将过表达RFX2后用二甲基亚砜（DMSO）或PY-6处理的A-549和NCI-H358细胞与活化的CD8 T细胞共培养。
RFX2在LUAD中显著下调。RFX2过表达增加了移植瘤内浸润的CD8 T细胞，并抑制了LUAD细胞的免疫逃逸、增殖和侵袭。RFX2富集于RASSF1启动子，并且RFX2通过与RASSF1启动子结合激活RASSF1转录。RASSF1敲低逆转了RFX2过表达抑制免疫逃逸的能力。RFX2缺失下调RASSF1，降低YAP磷酸化，从而影响Hippo通路以促进免疫逃逸。
LUAD中RFX2缺失下调RASSF1表达和YAP磷酸化，从而通过Hippo信号促进免疫逃逸。
