Zheng Hua-Chuan, Xue Hang, Wu Xin, Xu Hai-Lan, Zhao En-Hong, Cui Zheng-Guo
Department of Oncology and Experimental Center, The Affiliated Hospital of Chengde Medical University, Chengde, China.
Department of Pathology, Basic Medical College, Hebei North University, Zhangjiakou, China.
Front Oncol. 2022 Jun 7;12:918954. doi: 10.3389/fonc.2022.918954. eCollection 2022.
ING5 targets histone acetyltransferase or histone deacetylase complexes for local chromatin remodeling. Its transcriptional regulation and suppressive effects on gastric cancer remain elusive. Luciferase assay, EMSA, and ChIP were used to identify the cis-acting elements and trans-acting factors of the ING5 gene. We analyzed the effects of SAHA on the aggressive phenotypes of ING5 transfectants, and the effects of different ING5 mutants on aggressive phenotypes in SGC-7901 cells. Finally, we observed the effects of ING5 abrogation on gastric carcinogenesis. EMSA and ChIP showed that both SRF (-717 to -678 bp) and YY1 (-48 to 25bp) interacted with the promoter of and up-regulated ING5 expression in gastric cancer SRF-YY1-ING5-p53 complex formation. ING5, SRF, and YY1 were overexpressed in gastric cancer, (<0.05), and associated with worse prognosis of gastric cancer patients (<0.05). ING5 had positive relationships with SRF and YY1 expression in gastric cancer (<0.05). SAHA treatment caused early arrest at S phase in ING5 transfectants of SGC-7901 (<0.05), and either 0.5 or 1.0 μM SAHA enhanced their migration and invasion (<0.05). The wild-type and mutant ING5 transfectants showed lower viability and invasion than the control (<0.05) with low CDC25, VEGF, and MMP-9 expression. Gastric spontaneous adenocarcinoma was observed in Atp4b-cre; ING5, Pdx1-cre; ING5, and K19-cre; ING5 mice. ING5 deletion increased the sensitivity of MNU-induced gastric carcinogenesis. ING5 mRNA might be a good marker of gastric carcinogenesis, and poor prognosis. ING5 expression was positively regulated by the interaction of SRF-YY1-ING5-p53 complex within the ING5 promoter from -50 bp upstream to the transcription start site. ING5 deletion might contribute to the tumorigenesis and histogenesis of gastric cancer.
ING5靶向组蛋白乙酰转移酶或组蛋白去乙酰化酶复合物以进行局部染色质重塑。其对胃癌的转录调控和抑制作用仍不清楚。使用荧光素酶测定、电泳迁移率变动分析（EMSA）和染色质免疫沉淀（ChIP）来鉴定ING5基因的顺式作用元件和反式作用因子。我们分析了伏立诺他（SAHA）对ING5转染细胞侵袭性表型的影响，以及不同ING5突变体对SGC-7901细胞侵袭性表型的影响。最后，我们观察了ING5缺失对胃癌发生的影响。EMSA和ChIP表明，血清反应因子（SRF，-717至-678 bp）和阴阳1（YY1，-48至25 bp）均与ING5启动子相互作用，并在胃癌中上调ING5表达，形成SRF-YY1-ING5-p53复合物。ING5、SRF和YY1在胃癌中高表达（P<0.05），并与胃癌患者的不良预后相关（P<0.05）。在胃癌中，ING5与SRF和YY1的表达呈正相关（P<0.05）。SAHA处理导致SGC-7901的ING5转染细胞在S期早期停滞（P<0.05），0.5或1.0 μM的SAHA均增强了它们的迁移和侵袭能力（P<0.05）。野生型和突变型ING5转染细胞的活力和侵袭能力均低于对照组（P<0.05），且细胞周期蛋白依赖性激酶25（CDC25）、血管内皮生长因子（VEGF）和基质金属蛋白酶9（MMP-9）的表达较低。在Atp4b-cre; ING5、Pdx1-cre; ING5和K19-cre; ING5小鼠中观察到胃自发性腺癌。ING5缺失增加了N-甲基-N-亚硝基脲（MNU）诱导的胃癌发生的敏感性。ING5 mRNA可能是胃癌发生和不良预后的良好标志物。ING5启动子从转录起始位点上游-50 bp处的SRF-YY1-ING5-p53复合物相互作用对ING5表达起正调控作用。ING5缺失可能有助于胃癌的肿瘤发生和组织发生。
