RNA m6A甲基化阅读蛋白YTHDF2对食管鳞癌细胞放射敏感性的作用与机制研究

Objective: To primarily explore the roles and mechanisms of YTHDF2, the RNA m6A methylation reader protein, in regulating the radiosensitivity of esophageal squamous carcinoma (ESCC) cells. Methods: Recombinant adenovirus and siRNA were used to up/down-regulate YTHDF2 expression in human ESCC cell line TE-1 and KYSE-30, respectively. The effects and mechanisms of YTHDF2 on radiosensitivity of ESCC cells were investigated by colonogenic, apoptosis, DNA damage, and ROS assays. Results: After exposed to 4 Gy X-ray irradiation, the total m6A level in TE-1 cells was determined. And it was showed that total m6A in TE-1 was increased and then decreased within 24h. The mRNA level and protein level of YTHDF2 also showed a similar tendency. Compared with the control group, after overexpression of YTHDF2, the clone formation ability of TE-1 was obviously enhanced (TE-1:SER=0.887222) and the apoptosis rates were decreased (TE-1:P=0.0052), the level of ROS was decreased (TE-1:P=0.0117), and the DNA damage was reduced. And vice versa. Conclusion: The RNA m6A methylated reader protein YTHDF2 was involved in the regulation of radiosensitivity in ESCC cells. The mechanism might be related to its involvement in the regulation of radiation-induced apoptosis, ROS generation and DNA damage in ESCC cells. However, the specific molecular mechanisms involved were remain to be further investigated in depth.