转录因子 TEAD4 通过激活 PKMYT1 促进胃癌细胞的糖酵解和增殖。

Transcription factor TEAD4 facilitates glycolysis and proliferation of gastric cancer cells by activating PKMYT1.

机构信息

Department of Oncology, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou, 363000, China.

出版信息

Mol Cell Probes. 2023 Dec;72:101932. doi: 10.1016/j.mcp.2023.101932. Epub 2023 Nov 2.

DOI:10.1016/j.mcp.2023.101932

PMID:37729973

Abstract

BACKGROUND

Gastric cancer (GC) ranks third for cancer deaths worldwide, and glycolysis is a hallmark of several cancers, including GC. TEAD4 plays a role in establishing an oncogenic cascade in cancers, including GC. Whether TEAD4 can influence the glycolysis of GC cells remains uncovered. Hence, this study attempted to investigate the impact on glycolysis of GC cells by TEAD4.

METHODS

By using bioinformatics analysis, differentially expressed mRNAs were screened, and downstream regulatory genes were predicted. Expression levels of TEAD4 and PKMYT1 were assessed by qRT-PCR. The binding sites between TEAD4 and PKMYT1 were predicted by the JASPAR database, meanwhile their modulatory relationship was confirmed through dual-luciferase assay and chromatin Immunoprecipitation (ChIP). Cell viability and proliferation were assayed via CCK-8 and colony formation assays. Glycolysis was measured by assaying extracellular acidification rate, oxygen consumption rate, and production of pyruvic acid, lactate, citrate, and malate. Expression levels of proteins (HK-2 and PKM2) related to glycolysis were assessed by Western blot.

RESULTS

TEAD4 was upregulated in GC tissues and cells. TEAD4 knockdown substantially repressed glycolysis and proliferation of GC cells. PKMYT1, the target gene downstream of TEAD4, was identified via bioinformatics prediction, and its expression was elevated in GC. Dual-luciferase and ChIP assay validated the targeted relationship between the promoter region of PKMYT1 and TEAD4. As revealed by rescue experiments, the knockdown of TEAD4 reversed the stimulative effect on GC cell glycolysis and proliferation by forced expression of PKMYT1.

CONCLUSION

TEAD4 activated PKMYT1 to facilitate the proliferation and glycolysis of GC cells. TEAD4 and PKMYT1 may be possible therapeutic targets for GC.

摘要

背景

胃癌（GC）是全球癌症死亡的第三大原因，而糖酵解是包括 GC 在内的几种癌症的标志。TEAD4 在包括 GC 在内的癌症中发挥作用，建立致癌级联。TEAD4 是否会影响 GC 细胞的糖酵解仍未被发现。因此，本研究试图探讨 TEAD4 对 GC 细胞糖酵解的影响。

方法

通过生物信息学分析筛选差异表达的 mRNAs，并预测下游调控基因。通过 qRT-PCR 评估 TEAD4 和 PKMYT1 的表达水平。使用 JASPAR 数据库预测 TEAD4 和 PKMYT1 之间的结合位点，同时通过双荧光素酶报告基因检测和染色质免疫沉淀（ChIP）实验验证其调节关系。通过 CCK-8 和集落形成实验检测细胞活力和增殖。通过测定细胞外酸化率、耗氧量和丙酮酸、乳酸、柠檬酸和苹果酸的产生来测量糖酵解。通过 Western blot 检测与糖酵解相关的蛋白质（HK-2 和 PKM2）的表达水平。

结果

TEAD4 在 GC 组织和细胞中上调。TEAD4 敲低显著抑制 GC 细胞的糖酵解和增殖。通过生物信息学预测，鉴定出 TEAD4 的下游靶基因 PKMYT1，其在 GC 中表达上调。双荧光素酶和 ChIP 实验验证了 PKMYT1 启动子区与 TEAD4 之间的靶向关系。通过挽救实验表明，强制表达 PKMYT1 逆转了 TEAD4 敲低对 GC 细胞糖酵解和增殖的刺激作用。