Analysis of differential membrane proteins related to matrix stiffness-mediated metformin resistance in hepatocellular carcinoma cells.

IF 2.1 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS Proteome Science Pub Date : 2023-09-22 DOI:10.1186/s12953-023-00216-7

Xiangyu Gao, Jiali Qian, Yang Zhang, Heming Wang, Jiefeng Cui, Yehong Yang

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Abstract

Background: Our previous work shows that increased matrix stiffness not only alters malignant characteristics of hepatocellular carcinoma (HCC) cells, but also attenuates metformin efficacy in treating HCC cells. Here, we identified differential membrane proteins related to matrix stiffness-mediated metformin resistance for better understand therapeutic resistance of metformin in HCC.

Methods: Differential membrane proteins in HCC cells grown on different stiffness substrates before and after metformin intervention were screened and identified using isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with the liquid chromatography-tandem mass spectrometry (LC-MS/MS), then bioinformatic analysis were applied to determine candidate membrane protein and their possible signaling pathway.

Results: A total of 5159 proteins were identified and 354 differential membrane proteins and membrane associated proteins, which might be associated with matrix stiffness-mediated metformin resistance were discovered. Then 94 candidate membrane proteins including 21 up-regulated protein molecules and 73 down-regulated protein molecules were further obtained. Some of them such as Annexin A2 (ANXA2), Filamin-A (FLNA), Moesin (MSN), Myosin-9 (MYH9), Elongation factor 2 (eEF2), and Tax1 binding Protein 3 (TAX1BP3) were selected for further validation. Their expressions were all downregulated in HCC cells grown on different stiffness substrates after metformin intervention. More importantly, the degree of decrease was obviously weakened on the higher stiffness substrate compared with that on the lower stiffness substrate, indicating that these candidate membrane proteins might contribute to matrix stiffness-mediated metformin resistance in HCC.

Conclusions: There was an obvious change in membrane proteins in matrix stiffness-mediated metformin resistance in HCC cells. Six candidate membrane proteins may reflect the response of HCC cells under high stiffness stimulation to metformin intervention, which deserve to be investigated in the future.

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肝细胞癌细胞中与基质硬度介导的二甲双胍耐药性相关的差异性膜蛋白的分析。

背景：我们之前的研究表明，基质硬度的增加不仅改变了肝细胞癌（HCC）细胞的恶性特征，而且削弱了二甲双胍治疗HCC细胞的疗效。在这里，我们鉴定了与基质硬度介导的二甲双胍耐药性相关的差异膜蛋白，以更好地了解二甲双胍在HCC中的治疗耐药性。方法：使用相对和绝对定量同量异位标签（iTRAQ）标记结合液相色谱-串联质谱法（LC-MS/MS）对二甲双胍干预前后生长在不同硬度基质上的HCC细胞中的差异膜蛋白进行筛选和鉴定，然后应用生物信息学分析来确定候选膜蛋白及其可能的信号通路。结果：共鉴定出5159种蛋白质，发现354种可能与基质硬度介导的二甲双胍耐药性有关的差异性膜蛋白和膜相关蛋白。然后进一步获得94个候选膜蛋白，包括21个上调蛋白分子和73个下调蛋白分子。其中一些如膜联蛋白A2（ANXA2）、Filamin-A（FLNA）、Moesin（MSN）、肌球蛋白9（MYH9）、延伸因子2（eEF2）和Tax1结合蛋白3（TAX1BP3）被选择用于进一步验证。二甲双胍干预后，在不同硬度基质上生长的HCC细胞中，它们的表达均下调。更重要的是，与低硬度基质相比，高硬度基质的降低程度明显减弱，表明这些候选膜蛋白可能有助于HCC中基质硬度介导的二甲双胍耐药性。结论：基质硬度介导的HCC细胞对二甲双胍的耐药性使细胞膜蛋白发生明显变化。六种候选膜蛋白可能反映HCC细胞在高硬度刺激下对二甲双胍干预的反应，值得在未来进行研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Proteome Science 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

审稿时长

4.5 months

期刊介绍： Proteome Science is an open access journal publishing research in the area of systems studies. Proteome Science considers manuscripts based on all aspects of functional and structural proteomics, genomics, metabolomics, systems analysis and metabiome analysis. It encourages the submissions of studies that use large-scale or systems analysis of biomolecules in a cellular, organismal and/or environmental context. Studies that describe novel biological or clinical insights as well as methods-focused studies that describe novel methods for the large-scale study of any and all biomolecules in cells and tissues, such as mass spectrometry, protein and nucleic acid microarrays, genomics, next-generation sequencing and computational algorithms and methods are all within the scope of Proteome Science, as are electron topography, structural methods, proteogenomics, chemical proteomics, stem cell proteomics, organelle proteomics, plant and microbial proteomics. In spite of its name, Proteome Science considers all aspects of large-scale and systems studies because ultimately any mechanism that results in genomic and metabolomic changes will affect or be affected by the proteome. To reflect this intrinsic relationship of biological systems, Proteome Science will consider all such articles.