TRAF3IP2启动子区域的5mC和H3K9me3加速易位性肾细胞癌的进展。
5mC and H3K9me3 of TRAF3IP2 promoter region accelerates the progression of translocation renal cell carcinoma.
作者信息
Yang Lei, Chen Yi, Liu Ning, Lu Yanwen, Li Xin, Ma Wenliang, Gan Weidong, Li Dongmei
机构信息
Immunology and Reproduction Biology Laboratory & State Key Laboratory of Analytical Chemistry for Life Science, Medical School, Nanjing University, Nanjing, 210093, Jiangsu, China.
Jiangsu Key Laboratory of Molecular Medicine, Nanjing University, Nanjing, 210093, Jiangsu, China.
出版信息
Biomark Res. 2022 Jul 27;10(1):54. doi: 10.1186/s40364-022-00402-3.
BACKGROUND
In our previous study, we found that lncRNA TRAF3IP2 antisense RNA 1 (TRAF3IP2-AS1) could play a critical role in the progression of NONO-TFE3 translocation renal cell carcinoma (NONO-TFE3 tRCC). However, the function of TRAF3IP2 (TRAF3 interacting protein 2), encoded by the complementary strand of TRAF3IP2-AS1, remains poorly understood in NONO-TFE3 tRCC.
METHODS
Immunohistochemistry, western blot, and qRT-PCR were undertaken to study the expression and clinical significance of TRAF3IP2 in Xp11.2 tRCC tissues and cells. The functions of TRAF3IP2 in tRCC were investigated by proliferation analysis, EdU staining, colony and sphere formation assay, Transwell assay, and apoptosis analysis. The regulatory mechanisms among TRAF3IP2, NOTCH1, and TRAF3IP2-AS1 were investigated by luciferase assay, RNA immunoprecipitation, western blot, methylated DNA Immunoprecipitation, and CRISPR/dCas9-based system.
RESULTS
The results showed that TRAF3IP2 was highly expressed in NONO-TFE3 tRCC tissues and cells, and the silence of TRAF3IP2 inhibited the proliferation, migration, and invasion of UOK109 cells which were derived from cancer tissue of patient with NONO-TFE3 tRCC. Mechanistic studies revealed that TRAF3IP2 functioned as a co-activator of NOTCH1 to activate the NOTCH1 pathway. Meanwhile, HNRNPK, DNMT1 and SETDB1 could be recruited by TRAF3IP2-AS1 to the promoter region of TRAF3IP2, which mediated 5-hydroxymethylcytosine (5mC) on DNA and trimethylated lysine 9 of histone H3 (H3K9me3) at transcriptional level to repress the expression of TRAF3IP2.
CONCLUSIONS
TRAF3IP2 functions as an oncogene in NONO-TFE3 tRCC progression and might serve as a novel target for NONO-TFE3 tRCC therapy.
背景
在我们之前的研究中，我们发现长链非编码RNA TRAF3IP2反义RNA 1（TRAF3IP2-AS1）在NONO-TFE3易位性肾细胞癌（NONO-TFE3 tRCC）的进展中起关键作用。然而，由TRAF3IP2-AS1互补链编码的TRAF3IP2（TRAF3相互作用蛋白2）在NONO-TFE3 tRCC中的功能仍知之甚少。
方法
采用免疫组织化学、蛋白质免疫印迹和qRT-PCR研究TRAF3IP2在Xp11.2 tRCC组织和细胞中的表达及临床意义。通过增殖分析、EdU染色、集落和球体形成试验、Transwell试验和凋亡分析研究TRAF3IP2在tRCC中的功能。通过荧光素酶试验、RNA免疫沉淀、蛋白质免疫印迹、甲基化DNA免疫沉淀和基于CRISPR/dCas9的系统研究TRAF3IP2、NOTCH1和TRAF3IP2-AS1之间的调控机制。
结果
结果显示，TRAF3IP2在NONO-TFE3 tRCC组织和细胞中高表达，沉默TRAF3IP2可抑制源自NONO-TFE3 tRCC患者癌组织的UOK109细胞的增殖、迁移和侵袭。机制研究表明，TRAF3IP2作为NOTCH1的共激活因子激活NOTCH1通路。同时，TRAF3IP2-AS1可将HNRNPK、DNMT1和SETDB1募集到TRAF3IP2的启动子区域，在转录水平介导DNA上的5-羟甲基胞嘧啶（5mC）和组蛋白H3赖氨酸9的三甲基化（H3K9me3）以抑制TRAF3IP2的表达。
结论
TRAF3IP2在NONO-TFE3 tRCC进展中起癌基因作用，可能成为NONO-TFE3 tRCC治疗的新靶点。
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