Hu Feike
Department of Orthopaedics, The 7th Medical Center of Chinese PLA General Hospital, No.5 Nanmencang, Dongsishitiao Road, Dongcheng District, Beijing, 100700, China.
Sci Rep. 2025 Jul 2;15(1):23187. doi: 10.1038/s41598-025-05221-8.
DOT1L, acting as a key epigenetic regulator, plays important roles in various biological processes, offering significant insights for the development of new cancer therapeutic strategies. This study investigates the impact of DOT1L on pyroptosis in osteosarcoma and its molecular mechanisms. Bioinformatics analysis was performed to identify genes associated with DOT1L. Western blotting was used to detect the expression levels of relevant proteins; flow cytometry was used to assess apoptosis in Saos-2 cells; transmission electron microscopy was used to observe the number of pyroptotic bodies in Saos-2 cells; subcutaneous xenograft experiments in nude mice were used to evaluate the progression of osteosarcoma; immunohistochemical staining was used to detect the expression of DOT1L, p-SYK, p-EGFR, p-SHP2, and NLRP3 in tumor tissues. Bioinformatics analysis revealed that DOT1L is associated with H3K79me2/3. Under hypoxic conditions and with cGAMP treatment, DOT1L promoted the expression of H3K79me2/3, SYK, EGFR, p-SYK, p-EGFR, p-STING, p-TBK1, p-IRF3, P53, NLRP3, IL-1β, GSDMD, and apoptosis in Saos-2 cells, while inhibiting the expression of p-SHP2 and SHP2. DOT1L mediated the SYK/EGFR/SHP2 signaling pathway to increase the number of pyroptotic bodies in Saos-2 cells. Additionally, DOT1L suppressed the progression of osteosarcoma and enhanced the expression of p-SYK, p-EGFR, and NLRP3 in tumor tissues, while inhibiting p-SHP2 expression. DOT1L suppressed the progression of osteosarcoma by modulating SYK/EGFR/P53 and SHP2-induced STING-NLRP3-pyroptosis signaling.
DOT1L作为一种关键的表观遗传调节因子，在各种生物学过程中发挥着重要作用，为新的癌症治疗策略的开发提供了重要见解。本研究调查了DOT1L对骨肉瘤细胞焦亡的影响及其分子机制。进行生物信息学分析以鉴定与DOT1L相关的基因。采用蛋白质免疫印迹法检测相关蛋白的表达水平；采用流式细胞术评估Saos-2细胞的凋亡情况；采用透射电子显微镜观察Saos-2细胞中焦亡小体的数量；采用裸鼠皮下异种移植实验评估骨肉瘤的进展情况；采用免疫组织化学染色检测肿瘤组织中DOT1L、p-SYK、p-EGFR、p-SHP2和NLRP3的表达。生物信息学分析显示DOT1L与H3K79me2/3相关。在缺氧条件下并经cGAMP处理后，DOT1L促进了Saos-2细胞中H3K79me2/3、SYK、EGFR、p-SYK、p-EGFR、p-STING、p-TBK1、p-IRF3、P53、NLRP3、IL-1β、GSDMD的表达及细胞凋亡，同时抑制p-SHP2和SHP2的表达。DOT1L介导SYK/EGFR/SHP2信号通路增加Saos-2细胞中焦亡小体的数量。此外，DOT1L抑制骨肉瘤的进展，并增强肿瘤组织中p-SYK、p-EGFR和NLRP3的表达，同时抑制p-SHP2的表达。DOT1L通过调节SYK/EGFR/P53和SHP2诱导的STING-NLRP3-焦亡信号通路抑制骨肉瘤的进展。
