TROAP在STAT3的幫助下促進腎透明細胞癌的增殖、遷移和轉移

2023年6月，江南大學附屬醫院泌尿外科;南京醫科大學第一附屬醫院泌尿外科;江南大學無錫醫學院 (Department of Urology, Affiliated Hospital of Jiangnan University, Wuxi 214122, China;Department of Urology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210008, China；Wuxi Medical College, Jiangnan University, Wuxi 214122, China) Jun Wang老師研究團隊在《International Journal of Molecular Sciences》上發表論文：

 “TROAP Promotes the Proliferation, Migration, and Metastasis of Kidney Renal Clear Cell Carcinoma with the Help of STAT3"

“TROAP在STAT3的幫助下促進腎透明細胞癌的增殖、遷移和轉移"

Abstract：

Kidney renal clear cell carcinoma (KIRC) is a subtype of renal cell carcinoma that threatens human health. The mechanism by which the trophinin-associated protein (TROAP)-an important oncogenic factor-functions in KIRC has not been studied. This study investigated the specific mechanism by which TROAP functions in KIRC. TROAP expression in KIRC was analyzed using the RNAseq dataset from the Cancer Genome Atlas (TCGA) online database. The Mann-Whitney U test was used to analyze the expression of this gene from clinical data. The Kaplan-Meier method was used for the survival analysis of KIRC. The expression level of TROAP mRNA in the cells was detected using qRT-PCR. The proliferation, migration, apoptosis, and cell cycle of KIRC were detected using Celigo, MTT, wound healing, cell invasion assay, and flow cytometry. A mouse subcutaneous xenograft experiment was designed to demonstrate the effect of TROAP expression on KIRC growth in vivo. To further investigate the regulatory mechanism of TROAP, we performed co-immunoprecipitation (CO-IP) and shotgun liquid chromatography-tandem mass spectrometry (LC-MS). TCGA-related bioinformatics analysis showed that TROAP was significantly overexpressed in KIRC tissues and was related to higher T and pathological stages, and a poor prognosis. The inhibition of TROAP expression significantly reduced the proliferation of KIRC, affected the cell cycle, promoted cell apoptosis, and reduced cell migration and invasion. The subcutaneous xenograft experiments showed that the size and weight of the tumors in mice were significantly reduced after TROAP-knockdown. CO-IP and post-mass spectrometry bioinformatics analyses revealed that TROAP may combine with signal transducer and activator of transcription 3 (STAT3) to achieve tumor progression in KIRC; this was verified by functional recovery experiments. TROAP may regulate KIRC proliferation, migration, and metastasis by binding to STAT3.

摘要：

腎透明細胞癌(KIRC)是一種嚴重威脅人類健康的腎細胞癌亞型。TROAP是一種重要的致癌因子，其在KIRC中的作用機制尚未得到研究。本研究探討了TROAP在KIRC中的具體作用機制。使用來自癌癥基因組圖譜(TCGA)在線數據庫的RNAseq數據集分析TROAP在KIRC中的表達。采用Mann-Whitney U檢驗從臨床資料中分析該基因的表達。采用Kaplan-Meier法進行KIRC的生存分析。采用qRT-PCR檢測細胞中TROAP mRNA的表達水平。采用Celigo、MTT、創面愈合、細胞侵襲試驗和流式細胞術檢測KIRC的增殖、遷移、凋亡和細胞周期。研究人員設計了小鼠皮下異種移植實驗來證明TROAP表達對KIRC體內生長的影響。為了進一步研究TROAP的調控機制，研究人員采用了共免疫沉淀(CO-IP)和霰彈槍液相色譜-串聯質譜(LC-MS)。tcga相關生物信息學分析顯示，TROAP在KIRC組織中顯著過表達，與高T及病理分期、預后差有關。抑制TROAP表達可顯著降低KIRC的增殖，影響細胞周期，促進細胞凋亡，減少細胞遷移和侵襲。皮下異種移植實驗表明，敲除troap后，小鼠腫瘤的大小和重量明顯減小。CO-IP和質譜后生物信息學分析顯示，TROAP可能與信號換能器和轉錄激活因子3 (STAT3)結合，在KIRC中實現腫瘤進展;功能恢復實驗證實了這一點。TROAP可能通過結合STAT3調控KIRC的增殖、遷移和轉移。

該論文中，對人KIRC細胞系786-O、ACHN和Caki-1的體外培養是使用Ausbian特級胎牛血清完成的。