Department of Radiology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai 200233, China.
Department of General Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai 200233, China.
Aging (Albany NY). 2021 Feb 1;13(4):5185-5196. doi: 10.18632/aging.202439.
In this study, we investigated the effect of a short deletion in the DNA-binding domain of STAT3 (STAT3) on the transcriptional activation of STAT3 target genes and its relationship with colon carcinogenesis. We used the CRISPR-CAS9 gene editing system to delete a short sequence encoding amino acids 400-411 in the DNA-binding domain (amino acid sequence: 317-567) from gene in SW480, SW620 and HCT116 colon cancer cells. ChIP sequencing analysis showed that STAT3 occupancy was significantly reduced in 1029 genes and significantly increased in 475 genes compared to wild-type STAT3. The mutation altered the DNA motifs recognized by STAT3 as compared to the wild-type STAT3. We observed a strong correlation between expression of the STAT3 target genes and the loss or gain of STAT3 binding to their promoters. CCK-8, wound healing, and TUNEL assays showed reduced proliferation, migration, and survival of SW480, SW620 and HCT-116 cells expressing STAT3 as compared to the corresponding controls. These findings demonstrate that a short deletion in the DNA-binding domain of STAT3 alters its genome-wide DNA-binding and transcriptional profile of STAT3-target proteins, and suppresses the growth, progression and survival of colon cancer cells.
在这项研究中，我们研究了 STAT3（STAT3）DNA 结合域中的短缺失对 STAT3 靶基因转录激活的影响及其与结肠癌发生的关系。我们使用 CRISPR-CAS9 基因编辑系统从 SW480、SW620 和 HCT116 结肠癌细胞中的基因中删除编码 DNA 结合域中 400-411 个氨基酸的短序列（氨基酸序列：317-567）。ChIP 测序分析表明，与野生型 STAT3 相比，STAT3 占据的位置在 1029 个基因中显著减少，在 475 个基因中显著增加。该突变改变了 STAT3 识别的 DNA 基序，与野生型 STAT3 相比。我们观察到 STAT3 靶基因的表达与 STAT3 与其启动子结合的缺失或获得之间存在很强的相关性。CCK-8、划痕愈合和 TUNEL 测定表明，与相应对照相比，表达 STAT3 的 SW480、SW620 和 HCT-116 细胞的增殖、迁移和存活减少。这些发现表明，STAT3 DNA 结合域中的短缺失改变了其全基因组 DNA 结合和 STAT3 靶蛋白的转录谱，并抑制了结肠癌细胞的生长、进展和存活。
