上尿路尿路上皮癌(UTUC),包括肾,骨盆,输尿管癌,在台湾发病率很高,这与西方国家不同。因此,阐明UTUC生长和转移的潜在机制势在必行。探讨miR-145-5p在UTUC中的功能,我们用miR-145-5p模拟物转染BFTC909细胞系,并通过二维聚丙烯酰胺凝胶电泳分析蛋白质水平的差异.实时聚合酶链反应和蛋白质印迹分析用于分析5-氨基咪唑-4-甲酰胺核糖核苷酸甲酰基转移酶/肌醇单磷酸环水解酶(ATIC)信使RNA和蛋白质水平。进行双荧光素酶测定以鉴定ATIC中miR-145-5p的靶标。细胞转染miR-145-5p和ATIC表达对细胞增殖的影响,迁移,并对入侵进行了评估。miR-145-5p下调ATIC蛋白表达。ATIC高表达与肿瘤分期有关,转移,复发,UTUC患者预后不良。细胞功能检测显示ATIC敲低抑制细胞增殖,迁移,和UTUC细胞的侵袭能力。相比之下,miR-145-5p影响增殖,迁移,通过直接靶向ATIC的3'-非翻译区,UTUC细胞的侵袭能力。此外,我们使用RNA测序和独创性通路分析来鉴定ATIC调控的可能的下游基因,发现miR-145-5p调控纤连蛋白1,Slug,细胞周期蛋白A2,细胞周期蛋白B1,P57和干扰素通过ATIC诱导的跨膜1。ATIC可能是UTUC的有价值的预后预测因子和潜在的治疗靶点。
Upper tract urothelial carcinoma (UTUC), including renal, pelvic, and ureteral carcinoma, has a high incidence rate in Taiwan, which is different from that in Western countries. Therefore, it is imperative to elucidate the mechanisms underlying UTUC growth and metastasis. To explore the function of miR-145-5p in UTUC, we transfected the BFTC909 cell line with miR-145-5p mimics and analyzed the differences in protein levels by performing two-dimensional polyacrylamide gel electrophoresis. Real-time polymerase chain reaction and Western blot analysis were used to analyze 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/inositol monophosphate cyclohydrolase (ATIC) messenger RNA and protein levels. A dual-luciferase assay was performed to identify the target of miR-145-5p in ATIC. The effects of miR-145-5p and ATIC expression by cell transfection on cell proliferation, migration, and invasion were also assessed. miR-145-5p downregulated ATIC protein expression. High ATIC expression is associated with tumor stage, metastasis, recurrence, and a poor prognosis in patients with UTUC. Cell function assays revealed that ATIC knockdown inhibited the proliferation, migration, and invasive abilities of UTUC cells. In contrast, miR-145-5p affected the proliferation, migration, and invasive abilities of UTUC cells by directly targeting the 3\'-untranslated regions of ATIC. Furthermore, we used RNA sequencing and Ingenuity Pathway Analysis to identify possible downstream genes regulated by ATIC and found that miR-145-5p regulated the protein levels of fibronectin 1, Slug, cyclin A2, cyclin B1, P57, and interferon-induced transmembrane 1 via ATIC. ATIC may be a valuable predictor of prognosis and a potential therapeutic target for UTUC.