Zhengang Chen, Liu Huang, Xiaofeng Ji, Ranran Chen, Jiying Zhu
{"title":"Hexanoic Acid Production from Chinese Cabbage Waste Driven by In Situ Lactic Acid Pre-Fermentation: Effect of pH.","authors":"Zhengang Chen, Liu Huang, Xiaofeng Ji, Ranran Chen, Jiying Zhu","doi":"10.1007/s12010-025-05334-w","DOIUrl":"https://doi.org/10.1007/s12010-025-05334-w","url":null,"abstract":"<p><p>Chain elongation (CE) with lactic acid as electron donor is an important way for the biosynthesis of hexanoic acid, and pH is crucial for the carbon flow direction in the CE process. In this study, a high concentration of lactic acid was achieved through the pre-fermentation of Chinese cabbage wastes (CCW), and the effects of pH on the metabolic flow of lactic acid and hexanoic acid yield during CE reaction were investigated. The coexistence of Lactobacillus, Caproiciproducens, and Acinetobacter at pH 5.5 facilitated the CE process driven by lactic acid, obtaining the highest hexanoic acid yield of 5.01 ± 0.01 g COD/L. At pH 6.0, the high abundance of propionic acid-producing bacteria, such as Blautia and Succiniclasticum, converted lactic acid to propionic acid via the acrylate pathway, resulting in low selectivity of hexanoic acid. Without pH control, acetic acid was the main product due to the low pH at the initial stage of fermentation.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Improving the Accumulation and Solubility of Human Interferon-γ (hIFN-γ) in Escherichia coli: A Fusion Protein-Based Method and Network Pharmacology Analysis.","authors":"Narjes Akbari, Raheem Haddad, Reza Heidari-Japelaghi, Nematollah Gheibi","doi":"10.1007/s12010-025-05331-z","DOIUrl":"https://doi.org/10.1007/s12010-025-05331-z","url":null,"abstract":"<p><p>Using a network pharmacology-based method, some differentially expressed genes (DEGs) were detected in the colon cancer cell line (HCT116) after treatment with the human interferon-γ (hIFN-γ). Moreover, several pathways including cell cycle, NOD-like receptor signaling pathway, and P53 signaling pathway were identified, indicating the inhibitory effect of IFN-γ on the growth and proliferation of HCT116 cells. To validate in silico results, the hIFN-γ was first produced in Escherichia coli strain Rosetta and its bioactivity was then analyzed by anticancer assay. The production of hIFN-γ was performed via the optimization of several effective factors and the fusion of hIFN-γ to elastin like-polypeptide (ELP) tag. The highest amount of hIFN-γ (3.87 ± 0.37% of total soluble protein) was obtained at 22 °C with OD<sub>600</sub> = 0.6 and IPTG = 0.25 after 3-h inoculation. Whereas, the highest level of the hIFN-γ-ELP, about 4.58 ± 0.14% of TSP, was observed after 6-h inoculation. Compared to the hIFN-γ, the amount of hIFN-γ-ELP accumulation in the form of soluble increased significantly by more than 18%, proposing the desirable effect of ELP on the accumulation and solubility of hIFN-γ. Furthermore, the hIFN-γ prohibited the growth and proliferation of the HCT116 cells and the highest level of inhibition of cell proliferation was found at a concentration of 32.00 pg/mL hIFN-γ after 72-h incubation. Anticancer activity of hIFN-γ was also confirmed through the expression analysis of Bax, p53, and Bcl-2, suggesting the cytotoxic role of hIFN-γ toward HCT116 cells via inducing apoptosis process and arresting cell cycle.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"TCF4 Promotes Neuroblastoma Proliferation and Inhibits Ferroptosis by Transactivating GPX4 Expression.","authors":"Yingming Wang, Qiang Gao, Xin Chen, Qian Dong, Ruihong Luan, Fujiang Li, Hongting Lu, Xianjun Zhou","doi":"10.1007/s12010-025-05329-7","DOIUrl":"https://doi.org/10.1007/s12010-025-05329-7","url":null,"abstract":"<p><p>Neuroblastoma, an aggressive pediatric malignancy, exhibits aberrant expression of transcription factors implicated in tumor progression. Here, we investigated the functional role of transcription factor 4 (TCF4) in neuroblastoma, focusing on its impact on cellular proliferation and ferroptosis-a regulated form of iron-dependent cell death, and elucidated the underlying molecular mechanism. Firstly, the expressions of TCF4 in neuroblastoma tissues and cell lines were analyzed, and the expressions of TCF4 mRNA and protein were significantly up-regulated. Functional analysis demonstrated that sh-TCF4 could significantly proliferate neuroblastoma cells, which was measured by CCK-8 kit, EdU staining, and clone formation experiments. Concurrently, TCF4 knockdown significantly elevated ROS accumulation and lipid peroxidation levels. Besides, sh-TCF4 decreased the levels of FTH1 and increased the TFR1 expression. Mechanistically, bioinformatic analysis using the JASPAR database predicted TCF4 binding sites within GPX4 promoter, a key ferroptosis regulator. ChIP and dual-luciferase reporter assays confirmed direct TCF4 occupancy and transcriptional activation of GPX4. Rescue experiments further validated the axis, as GPX4 overexpression abrogated the anti-proliferative and pro-ferroptotic effects induced by sh-TCF4. Collectively, the findings revealed TCF4 as a critical promoter of neuroblastoma growth and ferroptosis resistance, acting through direct up-regulation of GPX4. Targeting the TCF4-GPX4 axis may offer a novel therapeutic strategy to enhance ferroptosis sensitivity in neuroblastoma, warranting further preclinical exploration.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Feng Zhu, Yuxian Ji, Qian You, Qigang Dong, Yao Tang, Yu Zhang
{"title":"Asiaticoside Mitigates Chronic Obstructive Pulmonary Disease by Modulating TRIM27 Stability and Activating PGC-1α/Nrf2 Signaling.","authors":"Feng Zhu, Yuxian Ji, Qian You, Qigang Dong, Yao Tang, Yu Zhang","doi":"10.1007/s12010-025-05288-z","DOIUrl":"https://doi.org/10.1007/s12010-025-05288-z","url":null,"abstract":"<p><p>Chronic obstructive pulmonary disease (COPD) is an inflammatory lung disease and is pathologically associated with epithelial-mesenchymal transition (EMT) and mitochondrial dysfunction. Asiaticoside (AS) has shown significant anti-inflammatory effects in a variety of diseases. Herein, the pharmacological influences of AS in COPD were probed. COPD mice were exposed to cigarette smoke (CS), and BEAS-2B cells were treated with cigarette smoke extract (CSE) for in vitro studies. HE staining was performed to assess lung pathological alters. The role of AS on inflammation, apoptosis, EMT, and mitochondrial dysfunction was analyzed by ELISA assay, western blot, Flow cytometry, DCFH-DA staining, and JC-1 staining. TRIM27 m6A expression was measured by MeRIP assay. The relationship between YTHDF1, TRIM27, and PGC-1α was determined by Co-IP or RIP assays. AS treatment relieved CSE-triggered inflammation, apoptosis, EMT, and mitochondrial dysfunction in a dose-dependent manner. Mechanically, AS suppressed PGC-1α ubiquitination degradation by reducing TRIM27 level in an m6A-YTHDF1-dependent manner. As expected, the mitigatory effect of AS on CSE-triggered BEAS-2B cell damage was abrogated by TRIM27 addition. Further, TRIM27 addition abrogated the restoring effect of AS on CS-caused pulmonary pathological damage in COPD mice. AS alleviated COPD by activating PGC-1α/Nrf2 signaling through weakening TRIM27 stability in an m6A-YTHDF1-dependent manner.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144641419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shanshan Zhao, Shouwen Du, Yudong Guan, Hartmut Schlüter
{"title":"Efficient Detection of N-sialoglycans in N-glycomics Using Mass Spectrometry by nMatch.","authors":"Shanshan Zhao, Shouwen Du, Yudong Guan, Hartmut Schlüter","doi":"10.1007/s12010-025-05337-7","DOIUrl":"https://doi.org/10.1007/s12010-025-05337-7","url":null,"abstract":"<p><p>The structural instability and electronegativity of sialic acids make N-sialoglycan profiling challenging. We compared commonly used preparation strategies, including native, acetohydrazide-based derivatization, and permethylation, in N-glycomics with nMatch, an R-based bioinformatics tool that ensures full coverage of the detected N-glycans using mass spectrometry. We demonstrated that permethylation leads to efficient detection of not only N-sialoglycans but also other N-glycans and, therefore, provides a valuable reference for the selection of the appropriate N-glycan preparation strategy in future studies.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144635873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A Novel Peptide Derived from Casein Hydrolysates as a Growth Factor for Lactobacillus delbrueckii subsp. bulgaricus sp1.1.","authors":"Xinyue Hao, Xiaoning Song, Jing Ren, Olayemi Eyituoyo Dudu, Jingya Jiang, Jianhua Zeng, Song Wang, Huaxi Yi, Lanwei Zhang, Pimin Gong","doi":"10.1007/s12010-025-05339-5","DOIUrl":"https://doi.org/10.1007/s12010-025-05339-5","url":null,"abstract":"<p><p>Milk proteins are traditionally recognized as nitrogen sources that support lactic acid bacteria (LAB) proliferation. Notably, oligopeptides may serve as the primary nitrogen source, and have the potential of enhancing the resistance of microbial strains to acidic environments. In this study, the impact of various casein hydrolysates on the proliferation of L. delbrueckii subsp. bulgaricus sp1.1 and the composition of peptides was studied. This study unveiled a novel peptide, KEGIHAQ, with a significant capacity to enhance both acid tolerance and proliferation of LAB. Inclusion of 0.1% (w/v) KEGIHAQ increased the number of viable bacteria by 1.68 times compared with the control and attained 1.34 × 10<sup>9</sup> CFU/mL, and under acid stress, the number of viable bacteria was 1.41 times that of the control. Transcriptomic analysis revealed that KEGIHAQ was not directly assimilated by L. delbrueckii subsp. bulgaricus sp1.1; instead, it acted as an effector, which upregulated the dipeptide transporter protein DppA and amino acid metabolism genes. This facilitated dipeptide uptake, amino acid synthesis, energy generation, and various bacterial physiological activities, while enhancing bacterial acid resistance mechanisms. These results provided valuable insights in identifying target peptides for nitrogen sources in future LAB high-density cultures, whether through biosynthesis or chemical synthesis.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144615701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"hsa_circ_0004771-Targeted Adsorption of miR-429 Mediates ATP2A2 Expression to Attenuate Endoplasmic Reticulum Stress and Mitochondrial Dysfunction During Myocardial Infarction.","authors":"Ying Deng, Ya Wu, SongChun Liu, YiJun Hou","doi":"10.1007/s12010-025-05310-4","DOIUrl":"https://doi.org/10.1007/s12010-025-05310-4","url":null,"abstract":"<p><p>Circular RNAs (circRNAs) have emerged as key regulators of cardiovascular pathophysiology. This study intended to mechanistically dissect the role of hsa_circ_0004771 in MI pathogenesis. An in vivo MI mouse model was established, and mouse cardiomyocytes (HL-1 cells) were cultured under hypoxic conditions to construct an in vitro cardiomyocyte injury model. Cardiac function in mice was assessed; serum markers of myocardial injury were quantified. Pathological alterations and levels of apoptosis in myocardial tissues were examined. Key oxidative stress indicators and pro-inflammatory cytokines were detected. Cell viability, cytotoxicity, and apoptosis were evaluated. Endoplasmic reticulum stress (ERS)-related proteins and mitochondrial function were analyzed. Serum samples from acute myocardial infarction (AMI) patients were collected for the detection of hsa_circ_0004771, miR-429, and ATPase sarcoplasmic/endoplasmic reticulum Ca<sup>2+</sup> transporting 2(ATP2A2) mRNA expression levels. Furthermore, dual-luciferase reporter assays and RNA pull-down assays were utilized to validate the interactions between miR-429 and circ_0004771 or ATP2A2. Circ_0004771 expression was significantly downregulated in AMI patients and experimental models. Overexpression of circ_0004771 improved cardiac function, ameliorated myocardial injury, decreased infarct size, suppressed oxidative stress markers, and attenuated inflammatory cytokine levels in MI mice. In vitro studies demonstrated that circ_0004771 overexpression alleviated hypoxia-induced injury in HL-1 cardiomyocytes, concurrently mitigating ERS, mitochondrial dysfunction, and calcium overload. Mechanistically, circ_0004771 acted as a sponge of miR-429, thereby upregulating ATP2A2 expression. Inhibition of miR-429 attenuated hypoxia-induced cardiomyocyte damage, while miR-429 overexpression reversed the protective effects of circ_0004771. Silencing of ATP2A2 abrogated the therapeutic benefits of miR-429 inhibition. Circ_0004771 mitigates hypoxia-induced ERS, mitochondrial dysfunction, and calcium overload via the miR-429/ATP2A2 axis in MI.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144599029","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Khushali Upadhyay, Foram Patel, Elizabeth Robin, A V Ramachandran, Darshee Baxi
{"title":"In-vitro Analysis of Solanum virginianum L. Extract and Melatonin: Synergistic Induction of Apoptosis in MDA-MB-231 Breast Cancer Cells.","authors":"Khushali Upadhyay, Foram Patel, Elizabeth Robin, A V Ramachandran, Darshee Baxi","doi":"10.1007/s12010-025-05335-9","DOIUrl":"https://doi.org/10.1007/s12010-025-05335-9","url":null,"abstract":"<p><p>Solanum virginianum (Sv) herb finds widespread usage in various medical systems, notably it is a component of the traditional herbal formulation \"Dashamula\". Melatonin is widely acknowledged as a chemical and a cell defender owing to its anti-oxidative and immunomodulatory properties. Despite extensive references in traditional medicine, systematic evaluations of Solanum virginianum anticancer properties are scarce. Combination therapy has emerged as a promising strategy for tackling drug-resistant cancers, inhibiting tumour progression, and enhancing therapeutic efficacy. This study aims to investigate the anticancer effects of Solanum virginianum plant extract and melatonin, individually and in combination, on breast cancer cells. The anticancer potential of individual treatments, as well as combinations of Solanum virginianum leaf extract and melatonin, was evaluated using cell migration inhibition, clonogenic assay, DNA fragmentation assay, nuclear morphology study, flow cytometry (FACS) analysis, and gene expression investigation. The methanolic leaf extracts of Solanum virginianum demonstrated significant anti-proliferative effects on MDA-MB-231 breast cancer cells. Furthermore, the combination of Solanum virginianum methanolic leaf extract and melatonin exhibited anti-migratory and anti-tumorigenic properties. This was further confirmed by gene expression studies related to both intrinsic and extrinsic apoptotic pathways. Notably, In combination groups, there was an increased expression of apoptotic genes (CASP3 (p < 0.001), CASP8 (p < 0.05), CASP9 (p < 0.05), BAX (p < 0.01) and anti-inflammatory genes (IL4, IL10), along with a decreased expression of the anti-apoptotic gene (BCL2) and metastatic genes (MMP2, MMP9). Overall, present study is the first comprehensive investigation into the potential of Solanum virginianum as an anticancer agent in conjunction with melatonin, highlighting the promising application of a combination approach for breast cancer treatment.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144599030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"METTL3-Mediated N6-Methyladenosine Modification Regulates the Progression of Diabetic Retinopathy.","authors":"Huaiyan Jiang, Wenzhong Fu, Yunmin Cai, Hongxia Xu","doi":"10.1007/s12010-025-05302-4","DOIUrl":"https://doi.org/10.1007/s12010-025-05302-4","url":null,"abstract":"<p><p>Diabetic retinopathy (DR) is a serious complication associated with diabetes, which may lead to diminished visual acuity or complete loss of sight. N6-methyladenosine (m6A) is recognized as the predominant modification present in eukaryotic mRNAs. However, the role of methyltransferase-like 3 (METTL3)-mediated m6A modification in DR still need further investigation. In our study, we constructed a DR cell model through the application of high glucose (HG) treatment on human retinal microvascular endothelial cells (hRMECs), and we found that METTL3 was downregulated for expression in the DR cell model. Mechanistically, we found that lncRNA MALAT1 was upregulated in hRMECs treated with HG due to the downregulation of METTL3 and subsequent reduction in m6A methylation. Functional experiments demonstrated that HG-induced upregulation of cell viability, endothelial-mesenchymal transition (EndMT), angiogenesis, and inflammatory factors were reversed by overexpressing METTL3, whereas these alleviating effects were neutralized by upregulation of MALAT1. Besides, inhibition of MALAT1 effectively attenuated retinal damage and inflammatory responses in streptozotocin (STZ)-induced DR mice. Furthermore, MALAT1 could act as a microRNA (miR)-23a-3p sponge to increase vascular endothelial growth factor A (VEGFA) expression. Reversal experiments indicated that knockdown of MALAT1 mediated DR alleviation effects were reversed by miR-23a-3p inhibitor or overexpression of VEGFA. In summary, our research findings indicated that silencing METTL3 reduced m6A modification of lncRNA MALAT1 and stabilized MALAT1 expression, thus promoted the growth, EndMT, angiogenesis, and inflammatory response of HG-induced hRMECs through the miR-23a-3p/VEGFA axis.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xinyue Bao, Jun Yan, Wanting Bai, Hao Ru, Chengzhi Yao, Qi Gao, Ruimin Gong, Junzheya Zhu, Jiebin Pan, Qi Sun
{"title":"Mendelian Randomization-Based Discovery of Novel Protein Biomarkers and Drug Targets in Colorectal Cancer: Validation Through Prognostic Modeling, Single-Cell Analysis, and In Vitro Cell Experiments.","authors":"Xinyue Bao, Jun Yan, Wanting Bai, Hao Ru, Chengzhi Yao, Qi Gao, Ruimin Gong, Junzheya Zhu, Jiebin Pan, Qi Sun","doi":"10.1007/s12010-025-05306-0","DOIUrl":"https://doi.org/10.1007/s12010-025-05306-0","url":null,"abstract":"<p><p>RNA modification plays a crucial role in biological processes. This study uses Mendelian randomization to investigate the relationship between RNAm-SNPs and CRC to identify potential protein markers and therapeutic targets. We analyzed RNAm-SNPs and CRC using RMVar and GWAS datasets. eQTL and pQTL analyses were performed to assess associations with gene expression and protein levels. Two-sample MR and summary-data-based MR identified candidate proteins. Single-cell expression analysis, prognostic model construction, protein-protein interaction studies, and druggability evaluations were conducted to identify cell-type enrichment and prioritize targets. Cell experiments further validated key genes in CRC. Six proteins (STX10, TBCA, GRIA4, PPT1, Sperm-associated antigen 2, and IL-21) were linked to CRC risk. These genes are primarily active in fibroblasts and epithelial cells in colon tumor tissue. Three proteins (GRIA4, IL-21, PPT1) are established targets for psychiatric and tumor disorders and may serve as therapeutic targets for CRC. Predictive modeling showed potential for clinical decision-making, and cell experiments confirmed TBCA as protective and GRIA4 as a risk factor in CRC. This study identified protein biomarkers associated with CRC risk, uncovering potential screening biomarkers and therapeutic targets, providing insight into the disease's molecular mechanisms.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582763","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}