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2023年6月，江南大學(xué)附屬醫(yī)院泌尿外科;南京醫(yī)科大學(xué)第一附屬醫(yī)院泌尿外科;江南大學(xué)無錫醫(yī)學(xué)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(tuán)隊(duì)在《International Journal of Molecular Sciences》上發(fā)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(jìn)腎透明細(xì)胞癌的增殖、遷移和轉(zhuǎn)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(xì)胞癌(KIRC)是一種嚴(yán)重威脅人類健康的腎細(xì)胞癌亞型。TROAP是一種重要的致癌因子，其在KIRC中的作用機(jī)制尚未得到研究。本研究探討了TROAP在KIRC中的具體作用機(jī)制。使用來自癌癥基因組圖譜(TCGA)在線數(shù)據(jù)庫的RNAseq數(shù)據(jù)集分析TROAP在KIRC中的表達(dá)。采用Mann-Whitney U檢驗(yàn)從臨床資料中分析該基因的表達(dá)。采用Kaplan-Meier法進(jìn)行KIRC的生存分析。采用qRT-PCR檢測細(xì)胞中TROAP mRNA的表達(dá)水平。采用Celigo、MTT、創(chuàng)面愈合、細(xì)胞侵襲試驗(yàn)和流式細(xì)胞術(shù)檢測KIRC的增殖、遷移、凋亡和細(xì)胞周期。研究人員設(shè)計(jì)了小鼠皮下異種移植實(shí)驗(yàn)來證明TROAP表達(dá)對(duì)KIRC體內(nèi)生長的影響。為了進(jìn)一步研究TROAP的調(diào)控機(jī)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(lián)質(zhì)譜(LC-MS)。tcga相關(guān)生物信息學(xué)分析顯示，TROAP在KIRC組織中顯著過表達(dá)，與高T及病理分期、預(yù)后差有關(guān)。抑制TROAP表達(dá)可顯著降低KIRC的增殖，影響細(xì)胞周期，促進(jìn)細(xì)胞凋亡，減少細(xì)胞遷移和侵襲。皮下異種移植實(shí)驗(yàn)表明，敲除troap后，小鼠腫瘤的大小和重量明顯減小。CO-IP和質(zhì)譜后生物信息學(xué)分析顯示，TROAP可能與信號(hào)換能器和轉(zhuǎn)錄激活因子3 (STAT3)結(jié)合，在KIRC中實(shí)現(xiàn)腫瘤進(jìn)展;功能恢復(fù)實(shí)驗(yàn)證實(shí)了這一點(diǎn)。TROAP可能通過結(jié)合STAT3調(diào)控KIRC的增殖、遷移和轉(zhuǎn)移。

該論文中，對(duì)人KIRC細(xì)胞系786-O、ACHN和Caki-1的體外培養(yǎng)是使用Ausbian特級(jí)胎牛血清完成的。