背景:转录因子FOXC1的活性增加导致靶基因转录升高,最终促进各种癌症类型的进展。然而,目前尚无关于FOXC1在肾细胞癌中作用的文献报道。
方法:通过使用RT-qPCR,免疫组织化学和蛋白质印迹,评估FOXC1mRNA和蛋白表达。利用功能增益实验来评估细胞的增殖和转移能力。建立裸鼠移植瘤模型,建立肺转移模型,观察细胞在生物体内的增殖和扩散。各种技术,包括生物分析,CHIP测定,荧光素酶测定,利用RT-qRCR和Western印迹实验来研究FOXC1如何在分子水平上促进ABHD5的转录。通过蛋白质印迹评估FOXC1对AMPK/mTOR信号通路的影响。
结果:FOXC1在RCC中下调,导致肾癌患者预后不良。进一步的实验表明,强制FOXC1表达显著抑制了RCC细胞的生长和细胞转移。机械上,FOXC1促进ABHD5转录激活AMPK信号通路抑制mTOR信号通路。最后,ABHD5的敲减恢复了FOXC1过表达诱导的细胞生长和转移抑制的抑制作用。
结论:一般来说,我们的研究表明,FOXC1通过促进ABHD5转录调节AMPK/mTOR信号通路发挥肿瘤抑制作用。FOXC1可作为RCC的诊断指标和潜在治疗重点。
BACKGROUND: Increased activity of the transcription factor FOXC1 leads to elevated transcription of target genes, ultimately facilitating the progression of various cancer types. However, there are currently no literature reports on the role of FOXC1 in renal cell carcinoma.
METHODS: By using RT-qPCR, immunohistochemistry and Western blotting, FOXC1 mRNA and protein expression was evaluated. Gain of function experiments were utilized to assess the proliferation and metastasis ability of cells. A nude mouse model was created for transplanting tumors and establishing a lung metastasis model to observe cell proliferation and spread in a living organism. Various techniques including biological analysis, CHIP assay, luciferase assay, RT-qRCR and Western blotting experiments were utilized to investigate how FOXC1 contributes to the transcription of ABHD5 on a molecular level. FOXC1 was assessed by Western blot for its impact on AMPK/mTOR signaling pathway.
RESULTS: FOXC1 is down-regulated in RCC, causing unfavorable prognosis of patients with RCC. Further experiments showed that forced FOXC1 expression significantly restrains RCC cell growth and cell metastasis. Mechanically, FOXC1 promotes the transcription of ABHD5 to activate AMPK signal pathway to inhibit mTOR signal pathway. Finally, knockdown of ABHD5 recovered the inhibitory role of FOXC1 overexpression induced cell growth and metastasis suppression.
CONCLUSIONS: In general, our study demonstrates that FOXC1 exerts its tumor suppressor role by promoting ABHD5 transcription to regulating AMPK/mTOR signal pathway. FOXC1 could serve as both a diagnostic indicator and potential treatment focus for RCC.