miR-204-5p缺失通过靶向食管鳞状细胞癌中的YWHAZ/PI3K/AKT通路促进肿瘤增殖、迁移和侵袭。
Loss of miR-204-5p Promotes Tumor Proliferation, Migration, and Invasion Through Targeting YWHAZ/PI3K/AKT Pathway in Esophageal Squamous Cell Carcinoma.
作者信息
Shen Zhimin, Chai Tianci, Luo Fei, Liu Zhun, Xu Hui, Zhang Peipei, Kang Mingqiang, Chen Sui
机构信息
Department of Thoracic Surgery, Fujian Medical University Union Hospital, Fuzhou 350001, People's Republic of China.
Key Laboratory of Ministry of Education for Gastrointestinal Cancer, Fujian Medical University, Fuzhou 350001, People's Republic of China.
出版信息
Onco Targets Ther. 2020 May 26;13:4679-4690. doi: 10.2147/OTT.S243215. eCollection 2020.
PURPOSE
MicroRNAs dysregulation has been confirmed in multiple malignancies. This paper reported the molecular mechanism of miR-204-5p in esophageal squamous cell carcinoma (ESCC).
METHODS
miR-204-5p expression in 30 ESCC tumor tissues and 10 normal tissues was downloaded from RNA-seq data. ESCC tissues/normal tissues of 97 ESCC patients were collected. TE-1 and KYSE510 cells were transfected by miR-204-5p mimic, inhibitor, siYWHAZ or their corresponding controls. The phenotype of cells was detected by CCK-8 assay, transwell experiment, and flow cytometry. Luciferase reporter gene assay and RNA-binding protein immunoprecipitation (RIP) were performed to verify the targeting relationship between miR-204-5p and YWHAZ. miR-204-5p and YWHAZ expression in tissues/cells was detected by qRT-PCR and Western blot. Xenograft tumor experiment was performed.
RESULTS
miR-204-5p expression was declined in ESCC patients and cells, which was indicated the poor outcome of patients. Compared with siNC group, TE-1 cells in miR-204-5p inhibitor group had higher OD450 value, less cell percentage in G1 phase, and more cell percentage in S phase, lower apoptosis percentage, and higher migration and invasion cell numbers. Moreover, KYSE510 cells of miR-204-5p mimic group showed lower OD450 value, more cell percentage in G1 phase and less cell percentage in S phase, higher apoptosis percentage, and lower migration and invasion cell numbers than control. YWHAZ was directly inhibited by miR-204-5p. Relative to siNC group, TE-1 cells of miR-inhibitor group exhibited higher YWHAZ protein expression, higher OD450 value, less cell percentage in G1 phase and more cell percentage in S phase, lower apoptosis percentage, higher migration and invasion cell numbers, and higher p-PI3K/PI3K and p-AKT/AKT protein expression, while siYWHAZ rescued the effects of miR-inhibitor. miR-204-5p up-regulation inhibited ESCC growth in vivo.
CONCLUSION
miR-204-5p inhibits ESCC progression by targeted inhibition of YWHAZ/PI3K/AKT.
目的
微小RNA（miRNA）失调已在多种恶性肿瘤中得到证实。本文报道了miR - 204 - 5p在食管鳞状细胞癌（ESCC）中的分子机制。
方法
从RNA测序数据中下载30例ESCC肿瘤组织和10例正常组织中miR - 204 - 5p的表达数据。收集97例ESCC患者的ESCC组织/正常组织。用miR - 204 - 5p模拟物、抑制剂、siYWHAZ或其相应对照转染TE - 1和KYSE510细胞。通过CCK - 8检测、Transwell实验和流式细胞术检测细胞表型。进行荧光素酶报告基因检测和RNA结合蛋白免疫沉淀（RIP）实验以验证miR - 204 - 5p与YWHAZ之间的靶向关系。通过qRT - PCR和蛋白质免疫印迹法检测组织/细胞中miR - 204 - 5p和YWHAZ的表达。进行异种移植瘤实验。
结果
ESCC患者和细胞中miR - 204 - 5p表达下降，提示患者预后不良。与siNC组相比，miR - 204 - 5p抑制剂组的TE - 1细胞OD450值更高，G1期细胞百分比更低，S期细胞百分比更高，凋亡百分比更低，迁移和侵袭细胞数量更多。此外，miR - 204 - 5p模拟物组的KYSE510细胞与对照组相比，OD450值更低，G1期细胞百分比更高，S期细胞百分比更低，凋亡百分比更高，迁移和侵袭细胞数量更低。YWHAZ被miR - 204 - 5p直接抑制。相对于siNC组，miR抑制剂组的TE - 1细胞YWHAZ蛋白表达更高，OD450值更高，G1期细胞百分比更低，S期细胞百分比更高，凋亡百分比更低,迁移和侵袭细胞数量更多，p - PI3K/PI3K和p - AKT/AKT蛋白表达更高，而siYWHAZ可挽救miR抑制剂的作用。miR - 204 - 5p上调可抑制ESCC在体内的生长。
结论
miR - 204 - 5p通过靶向抑制YWHAZ/PI3K/AKT抑制ESCC进展。
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