AATF is Overexpressed in Human Bladder Cancer and Regulates Chemo-Sensitivity Through Survivin.
AATF is Overexpressed in Human Bladder Cancer and Regulates Chemo-Sensitivity Through Survivin.
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AATF 在人类膀胱癌中过度表达并通过 Survivin 调节化疗敏感性
DOI：
10.2147/ott.s319734
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发表时间：
2021
影响因子：
4
通讯作者：
Dong Q
中科院分区：
文献类型：
--
作者：
Tan S;Fu L;Dong Q
目的细胞凋亡拮抗转录因子(AATF)的异常调控与人类肿瘤的发生密切相关。然而，它在人类膀胱癌(BC)中的作用尚不清楚。本研究旨在探讨AATF在膀胱癌中的临床意义和生物学作用。方法采用免疫组织化学方法检测107例膀胱癌组织中AATF蛋白的表达。对T24和5637细胞进行AATF质粒转染和小干扰RNA(SiRNA)敲除。采用CCK-8、集落形成、Annexin V/PI、JC-1染色和Western blotting等方法研究AATF在膀胱癌细胞中的生物学作用及其机制。结果AATF在膀胱癌组织中表达上调，且与T分期相关。对肿瘤数据库的分析显示，BC组织中AATF mRNA表达上调。癌症基因组图谱(TCGA)数据表明，AATF的高表达与患者存活率较低相关。Western blotting显示BC细胞系AATF蛋白表达高于正常膀胱移行上皮细胞系SV-Huc-1。CCK-8和集落形成实验表明，异位AATF表达上调了细胞的生长速度和集落数。CCK-8、Annexin V/PI、JC-1检测和Western blotting结果显示，AATF过表达降低顺铂敏感性，下调顺铂诱导的细胞凋亡，上调线粒体膜电位，降低细胞色素c和裂解PARP的表达。AATF siRNA下调则表现出相反的作用。机制上，AATF过表达上调了细胞周期蛋白E和Survivin的mRNA和蛋白水平。经Survivin抑制剂YM155治疗后，异位AATF诱导的顺铂敏感性降低/细胞凋亡被逆转。结论AATF在膀胱癌组织中高表达，并通过调控细胞周期蛋白E和Survivin促进肿瘤的恶性行为，提示AATF可作为膀胱癌的恶性标志物和潜在的治疗靶点。
Objective Dysregulation of apoptosis antagonizing transcription factor (AATF) has been reported to be closely associated with human cancers. However, its involvement in human bladder cancer (BC) remains unexplored. This study aimed to investigate the clinical significance and biological roles of AATF in human bladder cancers. Methods AATF protein expression was examined in 107 cases of bladder cancer tissues using immunohistochemistry. AATF plasmid transfection and small interfering RNA (siRNA) knockdown were performed in T24 and 5637 cell lines. CCK-8, colony formation, annexin V/PI, JC-1 staining, and Western blotting were carried out to investigate the biological roles and underlying mechanisms of AATF in bladder cancer cells. Results Our results showed that AATF expression was upregulated in human bladder cancer specimens and correlated with T stage. Analysis of the Oncomine database showed elevation of AATF mRNA in BC tissues. The Cancer Genome Atlas (TCGA) data suggested that high AATF expression correlated with poor patient survival. Western blotting showed that AATF protein expression was higher in BC cell lines compared to normal bladder transitional epithelial cell line SV-HUC-1. CCK-8 and colony assays showed that ectopic AATF expression upregulated cell growth rate and colony numbers. CCK-8, annexin V/propidium iodide (PI), JC-1 assays and Western blotting showed that AATF overexpression decreased cisplatin sensitivity, downregulated cisplatin-induced apoptosis and upregulated mitochondrial membrane potential, with decreased cytochrome c and cleaved-PARP expression. AATF siRNA knockdown showed the opposite effects. Mechanistically, AATF overexpression upregulated cyclin E and Survivin at both mRNA and protein levels. The decreased cisplatin sensitivity/apoptosis induced by ectopic AATF were reversed after treatment with Survivin inhibitor YM155. Conclusion Our results showed that AATF was overexpressed in human bladder cancers and promoted malignant behavior by regulating cyclin E and Survivin, indicating AATF could serve as a malignant biomarker and potential therapeutic target in BC.