Department of Otorhinolaryngology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
J Transl Med. 2022 Jun 6;20(1):255. doi: 10.1186/s12967-022-03468-7.
Lymphatic metastasis was an independent prognostic risk factor for hypopharyngeal carcinoma and was the main cause of treatment failure. The purpose of this study was to screen the differential genes and investigate the mechanism of lymphatic metastasis in hypopharyngeal carcinoma.
Transcriptome sequencing was performed on primary tumors of patients, and differential genes were screened by bioinformatics analysis. The expression of differential genes was verified by qRT-PCR, western-blotting and immunohistochemical, and prognostic value was analyzed by Kaplan-Meier and log-rank test and Cox's test. Next, FADU and SCC15 cell lines were used to demonstrate the function of differential genes both in vitro by EdU, Flow cytometry, Wound Healing and Transwell assays and in vivo by a foot-pad xenograft mice model. Proteomic sequencing was performed to screen relevant targets. In addition, in vitro and in vivo experiments were conducted to verify the mechanism of lymphatic metastasis.
Results of transcriptome sequencing showed that RAF1 was a significantly differential gene in lymphatic metastasis and was an independent prognostic risk factor. In vitro experiments suggested that decreased expression of RAF1 could inhibit proliferation, migration and invasion of tumor cells and promote apoptosis. In vivo experiments indicated that RAF1 could promote tumor growth and lymphatic metastasis. Proteomic sequencing and subsequent experiments suggested that LAGE1 could promote development of tumor and lymphatic metastasis, and was regulated by RAF1.
It suggests that RAF1 can promote lymphatic metastasis of hypopharyngeal carcinoma by regulating LAGE1, and provide a basis for the exploring of novel therapeutic target and ultimately provide new guidance for the establishment of intelligent diagnosis and precise treatment of hypopharyngeal carcinoma.
淋巴转移是下咽癌的独立预后危险因素，也是治疗失败的主要原因。本研究旨在筛选下咽癌淋巴转移的差异基因并探讨其机制。
对患者的原发肿瘤进行转录组测序，通过生物信息学分析筛选差异基因。采用 qRT-PCR、western-blotting 和免疫组化验证差异基因的表达，通过 Kaplan-Meier 和 log-rank 检验和 Cox 检验分析其预后价值。然后，使用 FADU 和 SCC15 细胞系通过 EdU、流式细胞术、划痕愈合和 Transwell 测定等体外实验以及足底异种移植小鼠模型等体内实验验证差异基因的功能。进行蛋白质组测序以筛选相关靶标。此外，还进行了体外和体内实验来验证淋巴转移的机制。
转录组测序结果表明，RAF1 是淋巴转移中显著差异的基因，是独立的预后危险因素。体外实验表明，RAF1 表达下调可抑制肿瘤细胞的增殖、迁移和侵袭，促进凋亡。体内实验表明，RAF1 可促进肿瘤生长和淋巴转移。蛋白质组测序及后续实验表明，LAGE1 可促进肿瘤的发生和淋巴转移，并受 RAF1 调节。
RAF1 可通过调节 LAGE1 促进下咽癌的淋巴转移，为探索新的治疗靶点提供了依据，并为下咽癌的智能诊断和精准治疗提供了新的指导。
