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

IF 2.3 3区 生物学 Q3 BIOCHEMICAL RESEARCH METHODS
Lifen Zhan , Wen Wu , Qiongling Yang , Huiqun Shen , Limin Liu , Renzhi Kang
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引用次数: 0

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.

转录因子TEAD4通过激活PKMYT1促进癌症细胞的糖酵解和增殖。
背景:癌症(GC)在全球癌症死亡人数中排名第三,糖酵解是包括GC在内的几种癌症的标志。TEAD4在建立包括GC在内的癌症致癌级联反应中发挥作用。TEAD4是否能影响GC细胞的糖酵解尚不清楚。因此,本研究试图研究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mRNA,并预测下游调控基因。通过qRT-PCR评估TEAD4和PKMYT1的表达水平。通过JASPAR数据库预测了TEAD4和PKMYT1之间的结合位点,同时通过双荧光素酶分析和染色质免疫沉淀(ChIP)证实了它们的调节关系。通过CCK-8和集落形成测定法测定细胞活力和增殖。通过测定细胞外酸化速率、耗氧速率以及丙酮酸、乳酸、柠檬酸盐和苹果酸的产生来测量糖酵解。通过蛋白质印迹评估与糖酵解相关的蛋白质(HK-2和PKM2)的表达水平。结果:TEAD4在GC组织和细胞中表达上调。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了TEAD4下游的靶基因PKMYT1,并在GC中提高了其表达。双荧光素酶和ChIP测定验证了PKMYT1启动子区和TEAD4之间的靶向关系。拯救实验表明,TEAD4的敲除逆转了PKMYT1强制表达对GC细胞糖酵解和增殖的刺激作用。结论:TEAD4激活PKMYT1促进GC细胞的增殖和糖酵解。TEAD4和PKMYT1可能是GC的可能治疗靶点。
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来源期刊
Molecular and Cellular Probes
Molecular and Cellular Probes 生物-生化研究方法
CiteScore
6.80
自引率
0.00%
发文量
52
审稿时长
16 days
期刊介绍: MCP - Advancing biology through–omics and bioinformatic technologies wants to capture outcomes from the current revolution in molecular technologies and sciences. The journal has broadened its scope and embraces any high quality research papers, reviews and opinions in areas including, but not limited to, molecular biology, cell biology, biochemistry, immunology, physiology, epidemiology, ecology, virology, microbiology, parasitology, genetics, evolutionary biology, genomics (including metagenomics), bioinformatics, proteomics, metabolomics, glycomics, and lipidomics. Submissions with a technology-driven focus on understanding normal biological or disease processes as well as conceptual advances and paradigm shifts are particularly encouraged. The Editors welcome fundamental or applied research areas; pre-submission enquiries about advanced draft manuscripts are welcomed. Top quality research and manuscripts will be fast-tracked.
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