mTOR/HK2信号的激活减轻了结直肠细胞中PYCR2缺失的影响。

IF 2.7 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2025-01-10 DOI:10.1016/j.tice.2025.102729

Li Chen , Yuan Yuan , Nian Zhang, Qianqian Huang, Yu Zhou

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引用次数: 0

摘要

背景：结直肠癌（Colorectal cancer， CRC）是侵袭性恶性肿瘤之一。研究表明糖酵解促进结直肠癌细胞增殖，PYCR2通过影响细胞糖酵解参与癌症进展。此外，PYCR2在结直肠癌细胞系中表达上调，可影响细胞自噬。方法：采用Si-PYCR2对结直肠癌细胞的PYCR2进行干扰，加入自噬抑制剂3-MA或mTOR激动剂MHY1485处理结直肠癌细胞。免疫荧光法检测LC3B的表达，Western blot法检测自噬蛋白和糖酵解蛋白的表达。采用XF96细胞外通量分析仪检测细胞ECAR和OCR，采用生化试剂盒检测细胞内葡萄糖消耗、乳酸分泌和ATP生成水平。结果：PYCR2在结直肠癌细胞系中表达上调。si-PYCR2干扰增强了细胞中LC3B的荧光强度，抑制了p62蛋白的表达，但增强了ATG5、ATG7和LC3-II/I蛋白的表达，表明结直肠癌细胞自噬水平增强。此外，PYCR2缺失也抑制细胞糖酵解和mTOR/HK2信号传导。然而，3-MA的加入导致细胞ECAR增加，OCR降低，葡萄糖消耗、乳酸和ATP产生水平增加，糖酵解蛋白（GLUT1、PGK1、ENO1、PKM2）表达增加，这表明细胞的糖酵解增强。此外，MHY1485治疗不仅抑制了结直肠癌细胞的自噬，还增强了糖酵解。结论：干扰PYCR2可通过mTOR/HK2信号通路纠正结直肠癌细胞中自噬依赖性糖酵解水平。mTOR/HK2信号的激活减轻了结直肠细胞中PYCR2缺失的影响。

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Activation of mTOR/HK2 signaling mitigates effects of PYCR2 depletion in colorectal cells

Background

Colorectal cancer (CRC) is one of the aggressive malignant tumors. Studies have shown that glycolysis promotes the proliferation of colorectal cancer cells and that PYCR2 is involved in cancer progression by affecting cellular glycolysis. In addition, PYCR2 is upregulated in colorectal cancer cell lines and can affect cellular autophagy.

Methods

Si-PYCR2 was used to interfere with PYCR2 in colorectal cancer cells, and the cells were treated with the addition of autophagy inhibitor 3-MA or mTOR agonist MHY1485. The expression of LC3B was detected by immunofluorescence, and the expression of autophagy and glycolytic proteins was detected by Western blot. XF96 extracellular flux analyzer was used to detect the ECAR and OCR of the cells, and biochemical kits were used to detect the levels of glucose consumption, lactate secretion, and ATP production in the cells.

Results

PYCR2 expression was up-regulated in colorectal cancer cell lines. si-PYCR2 interference enhanced the fluorescence intensity of LC3B in the cells, inhibited the expression of p62 proteins but enhanced the expression of ATG5, ATG7, and LC3-II/I proteins, which indicated an enhanced level of autophagy in colorectal cancer cells. In addition, PYCR2 depletion also inhibited cellular glycolysis as well as mTOR/HK2 signaling. However, the addition of 3-MA resulted in an increase in cellular ECAR while a decrease in OCR, and an increase in the levels of glucose consumption, lactate and ATP production, as well as the expressions of glycolytic proteins (GLUT1, PGK1, ENO1, PKM2), which suggested the glycolysis of cells was enhanced. In addition, MHY1485 treatment not only inhibited autophagy but also enhanced glycolysis in colorectal cancer cells.

Conclusion

Interference with PYCR2 corrected autophagy-dependent glycolysis levels in colorectal cancer cells via mTOR/HK2 signaling. Activation of mTOR/HK2 signaling mitigated the effects of PYCR2 depletion in colorectal cells.

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.